operation manual sylab

Operation Manual

IceCube 14S VAL

Operator Manual, Version 1.29 for Software Version 1.21 Val and up

SY-LAB Geräte GmbH., Tullnerbachstrasse 61-65, A 3011 Neupurkersdorf, Austria Tel +43 2231 622 52 –0, FAX +43 2231 621 93

Internet: http://www.sylab.com/, E-mail: [email protected], ©2000-2004 SY-LAB Geraete GmbH

http://www.sylab.com/

mailto:[email protected]

SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Operation Manual - IceCube 14S_VAL– Ver. 1.29 © 2004 SY-LAB Geräte GmbH.

All Rights Reserved - 2 -

Contents

Contents ____________________________________________________________________ 2

Warranty ____________________________________________________________________ 5

Specification _________________________________________________________________ 7 Freezer __________________________________________________________________________ 7

Power Connection _______________________________________________________________________ 7 Minimum PC Config. ____________________________________________________________________ 7 Freezing Chamber _______________________________________________________________________ 7 Capacity Options ________________________________________________________________________ 9 Software, Data Output____________________________________________________________________ 9

Liquid Nitrogen Dewar_____________________________________________________________ 9 Safety______________________________________________________________________ 10

General Precautions ______________________________________________________________ 10 Precaution (Liquid Nitrogen, etc.)___________________________________________________ 11 Precaution for Transportation______________________________________________________ 11 Packing Contents ________________________________________________________________ 12 Fuses___________________________________________________________________________ 13 Specific Notes____________________________________________________________________ 14

Installation and Check-out ____________________________________________________ 15 Setup Requirements ______________________________________________________________ 15 Site Preparation, Space and Clearance_______________________________________________ 15 Environment ____________________________________________________________________ 16 Unpacking ______________________________________________________________________ 16 IceCube Installation - Wiring and Hook-Up __________________________________________ 18

Variant A: ____________________________________________________________________________ 18 Variant B: ____________________________________________________________________________ 18 Variant C: ____________________________________________________________________________ 20 Variant D: ____________________________________________________________________________ 22 Connecting the LN2 Hose to Freezer and Supply Vessel ________________________________________ 23 Pt-100 Transport Protection_______________________________________________________________ 23 Turn ON/OFF and Check of the System _____________________________________________________ 25 Storage_______________________________________________________________________________ 25

Handling of Temperature Sensors___________________________________________________ 26 Freezing Chamber _______________________________________________________________ 28

Description ___________________________________________________________________________ 28 Available Freezing Racks __________________________________________________________ 29

Program Log-In _____________________________________________________________ 33 Description of Program Tools and Buttons ___________________________________________ 34

Menu Area____________________________________________________________________________ 36 Main Menu - File ____________________________________________________________________ 36

Edit description ___________________________________________________________________ 36 Open historical data ________________________________________________________________ 39




Process directory ________________________________________________________________ 39 Open last run ___________________________________________________________________ 39 Set source directory______________________________________________________________ 39 Print Diagram, Print Program & Events, Print both _____________________________________ 39 Close window __________________________________________________________________ 39

List of Available Tools_________________________________________________________ 42 Show description, Show program, Display Total View _________________________________________ 42

Freezing Program__________________________________________________________________ 43 Set data directory __________________________________________________________________ 43 Set program directory ______________________________________________________________ 44 Exit_____________________________________________________________________________ 44

Main Menu - Display _________________________________________________________________ 45 Total View _______________________________________________________________________ 45 Tracking View ____________________________________________________________________ 45 Settings__________________________________________________________________________ 45

Time axis______________________________________________________________________ 46 Temperature axis________________________________________________________________ 46 Time elapsed / remaining _________________________________________________________ 46 Actual program position __________________________________________________________ 46 Temperature value to track ________________________________________________________ 46 LNi pressure ___________________________________________________________________ 46

Main Menu - Options _________________________________________________________________ 47 Description of the various conditions and alarms _________________________________________ 48

Start Temperature reached condition_________________________________________________ 48 LNi pressure Alarm (not implemented on 14S) ________________________________________ 48 Allowed Temperature deviation ____________________________________________________ 49 Temperature reading invalid _______________________________________________________ 49 Critical power fail - awaiting user assistance __________________________________________ 49 Short time power fail_____________________________________________________________ 49 Freezing program finished ________________________________________________________ 50

Save Settings _____________________________________________________________________ 50 User Administration________________________________________________________________ 51

Info for the Administrator _________________________________________________________ 53 Language ________________________________________________________________________ 54

Main Menu - Version Info _____________________________________________________________ 55 Main Menu - Maintenance _____________________________________________________________ 55

IceWater Calibration _______________________________________________________________ 55 Communication ___________________________________________________________________ 57

Writing a Test Program ___________________________________________________________ 58 Test program for finding the crystallization temperature of a sample_______________________________ 58

Freezing program editor_____________________________________________________________ 59 Edit control - Program____________________________________________________________ 59

Edit Freezing program points ____________________________________________________ 59 Axis standard ________________________________________________________________ 62 Mouse button ________________________________________________________________ 62 Active point size______________________________________________________________ 62 Moving Screens and Zoom Functions _____________________________________________ 63

Running the Test Program________________________________________________________________ 64 Operation Panel ___________________________________________________________________ 68

Seeding _______________________________________________________________________ 68 Hold__________________________________________________________________________ 68 Next__________________________________________________________________________ 68 Load program/Stop ______________________________________________________________ 69

Writing the final program - Optimization (as an example)_____________________________________ 70 Special Function - Autoseeding _______________________________________________________ 74

Setting the Autoseeding Level _____________________________________________________ 74 Setting the Restart Level __________________________________________________________ 78

Seeding ____________________________________________________________________ 80




Auto(matic)Seeding_______________________________________________________________ 80 Maintenance ________________________________________________________________ 81

Leak Check _____________________________________________________________________ 81 Cleaning of the Freezing Chamber and Instrument ____________________________________ 81 Handling of Temperature Sensors___________________________________________________ 81

Settings on the PC ___________________________________________________________ 82 Disabling of Start-Up Error Message _______________________________________________________ 82 Disabling Request for Ctrl - Alt – Delete ____________________________________________________ 82 Disabling Power Save Options ____________________________________________________________ 83

Windows® XP-Professional ________________________________________________________ 83 Windows® 2000 Prof. _____________________________________________________________ 84

Freezer Software (Re-) Installation______________________________________________ 85 IceCube Series - License Agreement _________________________________________________ 85

Touch Monitor and TouchWare Setup ___________________________________________ 88 Program Installation______________________________________________________________ 88

Setup and Calibration of the Touch Monitor__________________________________________________ 91 ImageLoader________________________________________________________________ 94

Inserting Your Own LOGO ____________________________________________________ 95

Troubleshooting _____________________________________________________________ 96

Freezer and Accessories______________________________________________________ 102 Furniture ______________________________________________________________________ 102

Spare Parts 14S ____________________________________________________________ 103

Appendix __________________________________________________________________ 104 Statement of Conformity (220 - 240 V AC) __________________________________________ 104 Statement of Conformity (110 - 120 V AC) __________________________________________ 105 Monitor-Mounting Plate Assembly _________________________________________________ 106 Block Diagram - Software ________________________________________________________ 108




Warranty SY-LAB warrants for one year from the date of shipment each instrument manufactured to be free from defects in material and workmanship impairing its usefulness under normal use and service conditions except noted herein. Our liability under this warranty is limited to repair, servicing and adjustment, free of charge at our plant, of any instrument or defective parts, when returned prepaid to us, and which our examination discloses to have been defective. The purchaser is responsible for all transportation charges involving the shipment of materials for warranty repairs. Failure of any instrument or product due to operator error, improper installation, use of accessory parts from third party vendors (e.g. Liquid Nitrogen supply tubes, adaptors, etc.), unauthorized repair or alteration, failure of utilities, or environmental contamination will not constitute a warranty claim. The materials of construction used in SY-LAB instruments and other products were chosen after extensive testing and experience for their reliability and durability. However, these materials cannot be totally guaranteed against wear and/or decomposition by chemical action (corrosion) as a result of normal use. Repair parts are warranted to be free from defects in material and workmanship for 90 days from the date of shipment. No instrument shall be returned to us without our prior notice (return authorization no. required). SY-LAB shall be released from all obligations under its warranty in the event persons other than its own authorized service personnel make repairs or modifications unless such work is authorized in writing by SY-LAB. The obligations of this warranty will be limited under the following conditions: • Certain products sold by SY-LAB are the products of reputable manufacturers, sold under

their respective brand names or trade names. We, therefore, make no express or implied warranty as to such products. We shall use our best efforts to obtain from the manufacturer, in accordance with his customary practice, the repair or replacement of such of his products that may prove defective in workmanship or materials.

• If an instrument or product is found defective during the warranty period, replacement parts may, at the discretion of SY-LAB, be sent to the purchaser for installation, e.g. printed circuit boards, magnetic valves, temperature sensors, etc.

• Expendable items (e.g. o-rings, seal-rings, indicator lamps, fuses, etc.) are excluded from this warranty except for manufacturing defects. Such items, which perform satisfactorily during the first 45 days after the date of shipment, are assumed to be free of manufacturing defects.

• Due to the sensitivity of the temperature sensors (Pt-100), the warranty period on these devices will only be 12 weeks. This applies also to parts made out of PMMA (acrylic glass).

• Damage due to any static discharge. Before handling the temperature sensors, we advice the operator to discharge himself by touching shortly the grounded metal wall of the freezing chamber.

• Damage due to lightning, over voltage and/or heavy voltage/current fluctuation on the supply line.

Freezer: 1 year TFT Monitor: 1 year (5 years on Touch Monitor technology) Printer: 1 year from day of purchase

Please Note: For warranty purposes, it is necessary to mention model and serial number of TFT Monitor and Printer on YOUR invoice to the end user.

http://dict.leo.org/?p=2Ib6..&search=acrylic

http://dict.leo.org/?p=2Ib6..&search=glass




Please read the warranty instructions accompanying the monitor and printer. In most countries, monitor and printer can be repaired locally.




Specification Freezer

• Temperature Range: +40 to -180° Celsius (+104 to -292° Fahrenheit) • Manual and automatic control of heating and cooling • Pt-100 Temperature measurement with 2 resistance-thermometers (chamber- and

sample sensors) using 4 – wire technique, 1.5 mm Ǿ, 65 mm standard length or 100 mm (optional). Up to 2 additional sample sensors can be installed (at SYLAB)

• Electrical power-turn-OFF to motor, fan, heater and magnetic valves when lid is opened.

• Build-in safety valve – opens at approx. ( 50 psi ) 3,4 bar • Variable LN2 conn. from either side of the freezer with original tubing • Monitoring and alarm messaging of supply pressure • Maximum recommended working pressure: 1.5 bar (recomm. 0,5-0,8 bar) • 22 psi (recomm. 7 to 12 psi)

Note: Freezer can not be operated above 22 psi - error message shows up on screen

Cooling rates: 0.00-60°C/min Heating rates: 0.01-20 °C/min (in range -180°C to -50°C) 0.01-15 °C/min (in range - 50°C to +20°C) Display accuracy: 0.01°C Heating power: 750W Note: The above heating and cooling rates were measured with the freezing chamber empty.

Power Connection Freezer: (has to be specified on order) 220-240VAC (100-120VAC) 800VA (w/o monitor, printer and PC) TFT Touch screen: 1A max. / 100-240VAC, 40Watt max. - switching power supply

Minimum PC Config. Windows® 2000 or XPP, 1x serial, 1x USB, CD-Rom drive and 3 1/2" Floppy disk recommended.

Freezing Chamber (LxWxH in mm): 217 x 217 x 348 (LxWxH) in inch): 8,5 x 8,5 x 13,7 16,4 Liter (~4,3 gal.) chamber volume, Stainless Steel, hinged transparent glass cover, prepared for AutoSeeding, additional sample sensors possible, thermo fuse over heat protection Dimensions Freezer (LxWxH in mm): 540 x 420 x 520 (w/ PC system) (LxWxH in inch): 21,3 x 16,5 x 20,5 (w/ PC system Height with TFT monitor: 740 mm (~29,2 in.) Height with opened lid: 820 mm (~32,3 in.) Weight approx.: 29 kg (~63 lbs.)




Required area (LxWxH in mm): 850 x 550 x 870 (depending on configuration) (LxWxH in inch): 33,5 x 21,6 x 34,3 ( ---------------“----------------) Dimension Trolley (LxWxH in mm): 710 x 565 x 720 (LxWxH in inch): 28 x 22,3 x 28,4




Capacity Options 1080 (0, 25 / 0, 5 ml) straws or 480 (2ml) ampoules or 240 (5 ml) ampoules, 11 metal cassettes or several blood bags (number varies according on bag size and fill volume)

Software, Data Output Unlimited number of programs, 12.1“ TFT Touch Monitor (Variant B-D), program menu driven via Touch Monitor, removal of the heat of crystallization due to counter cooling, Hold and Next Ramp function, free adjustable start temperature, thermo sensor calibration routine, zoom function, Date- and Time integrated filename, color inkjet printer (option), program-, chamber- and sample curve.

Liquid Nitrogen Dewar Liquid Nitrogen supply vessel must comply with the local regulations for pressure vessels. Working pressure of 0, 3 to max. 1, 5 bar (7 to 22 psi), volume of 60 to 240 liters depending on the number of freezing cycles. Larger vessels require probably longer supply tubing (available through SY-LAB). For supply tubes longer than 1.3 m, new cooling and heating parameters might have to be acquired.

Note: To keep the instrument at its highest technical standard, SY-LAB reserves the right to make modifications / change specifications without the prior notice of the customer.




Safety General Precautions

• For your personal safety, read thoroughly the following sections before turning the instrument on for the first time.

• Pay attention to all notes, safety and warning messages mentioned in this manual and shown on the instrument.

• Always disconnect the instrument from the power line before attempting to open, clean or perform any service work. Do not use abrasives or cleaning sprays except when it is asked for in the manual. Use only a wet soft cloth for cleaning.

• Place the instrument on a flat and stable surface (table or desk). • Use only original Liquid Nitrogen supply tubes available through SY-LAB. Heating and

cooling parameters have been determined using SY-LAB supply tubes. SY-LAB will not take any responsibility for damages done to the plumbing or malfunction of the IceCube Freezer.

• The openings on the freezer housing are mainly used for cooling purposes. Do not block these openings otherwise, the instrument might get damaged due to over heating. Also, do not place the instrument close to heat sources or ventilation systems such as radiators, fans, etc.

• Before connecting the instrument to the mains, make sure that the voltage rating shown on the type-plate agrees with your main supply. If in doubt, contact your SY-LAB GmbH. representative or SY-LAB GmbH. Austria directly.

• Use only the original power supply cable. A grounded (ground potential) wall socket is required. If the wall socket does not fit the instruments supply cable, contact your local electrician.

• When using extension wires or distribution boxes, be sure not to exceed the maximum power rating of all instruments connected. This is normally about 15A AC.

• Never try to insert objects into any instrument openings. Electrical shock might be the result, which in turn might be harmful to life. Do not pour any liquid into any of the openings.

• With the exception of work asked for in this manual, never try to repair the instrument by yourself. Only authorized personnel must perform repair work.

• In all following cases, disconnect the instrument from the mains and call your service representative:

- Power supply cable or connector worn or defective - Liquid was spilled and entered the instrument - Instrument shows malfunction when used according to the operator manual.

Note: Use only accessories (as mentioned in the operator manual) intended to be used with this type of instrument.

- Instrument was dropped and/or housing is damaged. - Instrument shows strange behavior as compared to normal operation. - When instrument was stored for a long time.




Precaution (Liquid Nitrogen, etc.) • Liquid Nitrogen (LN2) is dangerous when it comes in contact with skin or when it gets into

the eyes. Use special care when handling LN2. For safety reasons, wear protective gloves, goggles and a leather apron.

• The new model freezers are built with metallic seals and must not be used with any additional seal rings.

• During a freezing cycle, the LN2supply tubing is getting very cold and if necessary, must only be touched with protective gloves. Ignoring this warning might result in severe burns and/or frostbites.

• Areas exposed to excess nitrogen gas (storage tanks, freezers etc.) must be well ventilated. Under normal circumstances, keeping a window area permanently open (at upper half of the room) will be inevitable. Nitrogen gas is not poisonous but it will displace the oxygen in the air. For safety reasons, the use of an oxygen monitor system (available trough SY-LAB GmbH.) is recommended. For badly ventilated areas, it will be mandatory. In any case, please contact your safety department for local regulations.

• A safety instruction booklet will accompany all supply Dewars delivered by SY-LAB. This booklet must be read and be understood before handling any Liquid Nitrogen equipment.

• Also, for safety reasons disconnect the instrument from the power line whenever maintenance work is performed on the instrument. Do not open the instrument while it is connected to the power-line. Electrical shock might result. We will void all warranty and will free ourselves from any obligations if our instruments are opened without the prior written consent of the manufacturer.

Precaution for Transportation During the transportation of the freezer, it is absolutely necessary to secure the Pt-100 temperature sensors. Use some Styrofoam pad, plastic tubing or similar and secure the sensors with tape to the inside wall of the freezing chamber. This keeps the sensors from bouncing around during transportation.

Caution: For normal operation of the freezer, the Pt-100s transport protection must be removed. Not doing so will lead to heavy temperature oscillations.

In addition, do not place any foreign parts or chamber furniture into the freezing compartment. Damage might occur if done so and will void any warranty. In case of printer or monitor failure, file a report to SY-LAB GmbH. and contact your local printer / monitor representative. Most printer / monitor manufactures have worldwide warranty service. Use only the original packing or pack separately when returned with the freezer. No return shipments will be accepted by SY-LAB without a Return Authorization Number (RAN). Do not throw the packing material away. Keep it for possible shipments in the future.




Packing Contents Please check the contents of the delivered goods. Obvious damage to packing or instrument must immediately be reported to the shipping agent. Any damage must be recorded on the delivery papers (invoice, etc.). If the shipment is accepted, then only with "reservation". In general, if no additional items were ordered, depending on the Variant, it should consist of the following: Variant A - Free standing freezing chamber - PC system provided by the customer

• IceCube freezer - basic unit • Power cable • Serial communication cable • Operator Manual • IceCube Freezer Software (CD) • Hardware dongle (key)

Necessary area for freezing chamber + PC system (L x W in mm): 540 x 400 (without PC) (L x W in inch): 21,3 x 15,7 (-----“------)

Variant B - Free standing freezing chamber with Touch Monitor

• IceCube freezer - basic unit • Power cable • Serial communication cable • Touch Monitor package • Monitor mount • Operator Manual • IceCube Freezer Software (CD) • Hardware dongle (key)

Necessary area for freezing chamber + PC system (L x W in mm): 740 x 400 (L x W in inch): 29 x 15,7 Variant C - Freezing chamber with Touch Monitor and standard PC-mount

• IceCube freezer - basic unit • Power cable • Serial communication cable • Touch Monitor package • Monitor mount • Standard PC mount (two arms) with straps • Operator Manual • IceCube Freezer Software (CD) • Hardware dongle (key)




Necessary area for freezing chamber + PC system (L x W in mm): 830 x 400 (L x W in inch): 32,7 x 15,7 Note: Max. PC - height: 390 mm (15,4 in.) Variant D - Freezing chamber with Touch Monitor and mini PC-mount

• IceCube freezer - basic unit • Power cable • Serial communication cable • Touch Monitor package • Monitor mount • Mini PC mount (single arm) • Operator Manual • IceCube Freezer Software (CD) • Hardware dongle (key)

Necessary area for freezing chamber + PC system (L x W in mm): 740 x 400 (L x W in inch): 29 x 15,7 Note: Max. PC - height: 300 mm (11,8 in.) Optional items for above Variants:

• Computer system originally packed • Printer • Accessory package - supply tubing, wrenches, etc. • Furniture f. freezing chamber - racks, seeding, spacer, etc. • Customized orders possible

Note: For monitor, printer and PC package contents, please consult the accompanying operator manual of the corresponding manufacturer.

Fuses Two Fuses protect the IceCube against overload. The fuses are located next to the main - power switch of the Freezer Electronic Box. Value: T 5 AL/250V Type: 20 x 5 mm fuse Caution: Disconnect the freezers power supply cable from the mains. Never change a fuse or perform internal work with the power cable plugged into the mains supply. Fuses




Specific Notes • Computer System from third party suppliers must comply with the territorial regulations. • The IceCube in the standard configuration comes with two temperature sensors. One

chamber- and one sample sensor. Optional, two more sensors can be added. However, this cannot be done in the field. So either you order the IceCube already with additional sensors or the freezers electronic box is sent to SY-LAB for up-grade.

• Liquid Nitrogen Vessels should only be filled to about 90% of their nominal filling volume. The free space above the liquid level is needed for gas building assuring continues supply of Nitrogen to the IceCube.

• Please note: Computer systems not purchased at SY-LAB must be configured prior to the installation into the IceCube. Besides the operating system, the freezer software as well as the Touch Monitor software must be installed. The instructions for loading the freezer program into your PC are found in this manual. The installation of the Touch Monitor is described in the monitor handbook.

• Since there will be no keyboard connected to your PC, deactivate "Hold On All Errors" in the computer BIOS. If this is not done, your PC system will stop when looking for the keyboard.

• Use only the original Liquid Nitrogen supply tubing from SY-LAB. The original supply tubing allows the placement of a supply vessel at either side of the IceCube freezer. Also, the heating and cooling parameters apply only to IceCube systems using the original supply tube.

• For supply vessels up to 60-liter content, the supply tube should be 1 meter long. For larger vessels, a 1.3 meter supply tube should be sufficient. If other length is required, pls. contact SY-LAB. Tubing Connections: 3/4" UNF Nut to G 1/4". Both ends use metallic sealing. No seal rings are required.

• Secure one or two wheels of the Liquid Nitrogen Vessel against movement. • Due to the very low temperature on the LN2 supply tubing and connections, water

condensation will occur. This is seen as white fog dropping to the floor. Water droplets in the surrounding air condense out and will sink to the ground. This is a normal process and has nothing to do with a leak in the LN2 system. Nitrogen gas is invisible and odorless.

• Never disconnect a cold supply tube from the supply vessel or the IceCube freezer. Wait until the supply tube has reached ambient temperature. If moisture due to condensation gets into the supply tube, it will lead to blockage of ice or to freeze the magnetic valve.

• Please note that for the setup of a Freezer, PC, Monitor and printer, 4 wall outlets (distribution block) are required.

• For normal operation of the freezer, the Pt-100s transport protection must be removed. Not doing so will lead to heavy temperature oscillations.




Installation and Check-out Setup Requirements

• Before connecting any instrument(s) to the power line, make sure the line voltage agrees with the voltage shown at the type plate of the Freezer and the Computer System.

• The instrument must be connected to a power source with proper grounding. Do not use extension cables without ground connections. Use only the original, from the manufacturer supplied power supply cables. Micro wave ovens, induction heaters, etc. could cause power disruption and noise on the power line. Consult your local electrician if problems are traced back to power line noise.

• Never remove any ground connections from the inside nor the outside of the instrument. • The Freezer must be connected to a common ground socket.

Any violation of this will be hazardous and dangerous to life. Also, all warranty will be void. The printer and monitor are using both a power supply of class II and need therefore only a 2-core wire.

• Use only the cables, originally supplied by the manufacturer. • Removal of any parts or opening the instrument might cause contact with high voltage.

Therefore, we urge anyone opening the instrument to disconnect the instrument from the power source. Any violation of this will be hazardous and dangerous to life.

• Any repair work or adjustments on an open instrument requiring the connection to the power line must be made through an authorized person knowing about the danger of high voltages.

• We want to point out that certain components such as, e.g. capacitors, might still be charged-up even though the instrument is disconnected from the power line.

• Never use any fuses of different type or value. Bad fuses must be discarded and be replaced with new ones.

• Operate the freezer only in rooms which are recommended by your local authorities (ventilation-, alarm system, etc.)

• If there is ever a reason to believe that the instrument does not operate safely or has become a safety hazard, immediately disconnect it from the power line, secure against unintentional use and call your local service representative or SY-LAB GmbH-Austria.

• Reasons to doubt safe operation:

• Instrument shows obvious damage • Instrument does not work anymore • After long storage in non-ideal area • After long and extreme transportation conditions.

Site Preparation, Space and Clearance An opening is provided on the backside of the freezer. It is used as an exhaust for the nitrogen gas. Position the IceCube in such a way that the cold exhaust gas can not damage any other equipment nearby. Keep at least a distance of 15 - 20 cm (6-8 inches) to other equipment. Additional requirements:

• Position the IceCube on a flat, horizontal and stable surface • The instrument must be accessible from all sides




• Do not place the instrument such that it is exposed to direct sunlight, extensive heat or cold, humidity and dust.

• You must be able to remove the power cord from the wall socket at any time. Do not block the access to the power socket.

• Do not use any wall sockets that can be turned ON or OFF by an extra switch or timer. Besides loosing data when automatically turned off, the sample material might be ruined as well.

• Do not use wall sockets where other heavy instruments such as motors, copy machines, ovens, etc. are connected. Such devices can shortly deplete the supply voltage when turned on.

• The computer system and its components should not be placed near big speaker systems, cordless- and mobile docking stations, etc.

• Freezer, computer and all other system components must be connected to the same power source having the same grounding potential. Potential differences due to bad grounding are potential sources of error and can lead to the destruction of any connected items. No warranty will be granted in such cases.

Environment The instrument will work without any difficulties under normal working conditions (15-35 °C / 60-95 °F). Nevertheless, in a humid environment, excessive condensation on the LN2-supply-tubing will occur and is normal. The environment should be such that 90% humidity (non condensing) is not exceeded. Very high humidity can lead to internal condensation and destruction of the instrument. Also, during transportation, it is vital that the instrument is given time to adjust to the surrounding temperatures before turned on, especially in the wintertime, when instruments are transferred from a cold to a warm environment or vice versa. Instruments on a trolley must be secured against movement. Use the roller-lock on the wheels to do so.

Unpacking Prior to the shipping, the instrument was carefully inspected and packed. Because of the size and weight, it needs special packing. Unpacking instructions are attached to the outside of the box. Check for any transport damages before opening the shipping box. If any damage is visible, immediately contact your shipping / insurance agent or better do not accept the shipment. If in doubt, make at least a note of the damage on the shipping papers. Take a picture as well. Check the contents of the package for completeness. Compare with the packing list, delivery note, etc.

If any items are missing, please contact immediately either your representative or SY-LAB Austria. Later reclamations will not be accepted. Always state Model Number and Serial Number of Freezer, Monitor, Printer, and so on. Do not return any items to SY-LAB without having a RAN (return authorization number). Note: Shipping- and insurance costs are not covered by SY-LAB. Do not dispose the packing material. You might need it for future shipments.




On the left, you see an original 14S shipping box. Size of box (LxWxH in mm): 670 x 520 x 580 (LxWxH in inch): 26,4 x 20,5 x 22,8 Open the box carefully from the top. Below the top, you will find another cardboard with handles.

Carefully pull the instrument out and pay attention that the instrument does not slip out on one of the sides.

Place the freezer on an appropriate location (see "Site Preparation and Clearance"). Depending on the Variant and options of your system, an accessory package will also be included (see "Packing Contents").




IceCube Installation - Wiring and Hook-Up Please consider the above mentioned environmental conditions. Place the IceCube on a flat and horizontal surface. Consider the stability of your table, laboratory trolley, etc. The IceCube weighs around 30 kg (~64 lb.). Note: Only trained personnel should perform the installation of the IceCube. Unpack monitor, printer, PC, etc. and inspect visually for any damage. The monitor stand was modified at SY-LAB (Variant B-D) and does not exactly look like the one shown in the monitor handbook. In the following section, we will go step-by-step through the instruments installation and hook-up procedure.

Variant A: (Free standing freezing chamber - PC system provided by the customer) In this configuration, only the freezing chamber is available. For the system to work, a PC system including monitor and keyboard are required. For hard copies, a printer will be useful.

The chamber comes with a power- and communication cable. Communication will be established between the freezers serial connection and a free COM Port on your PC. Use the supplied power cable and attach it to a wall outlet. Note: Make sure the

freezers power rating corresponds to your local supply (see Type-Plate). Thereafter, the IceCube Software can be installed (see " IceCube Series Software Installation).

Variant B: (Free standing freezing chamber and PC with Touch Monitor) This type of configuration includes already the Touch Monitor.

During normal operation, no extra keyboard or mouse will be needed. For hard copies, a printer will be useful.

In addition to the connections mentioned in Variant A, attach the stereo jack of the PC's line-out of the sound card to the line-input of the monitor (not on all units available). Also, attach the monitors Touch-Screen




USB-cable to one of the USB ports of your PC. As already mentioned above, after a successful installation of the operating system and the IceCube program incl. Touch Ware, etc., the keyboard and mouse will not be needed anymore.

Generally, the freezer series 14S will come in a basic configuration (without Monitor and PC support). Depending on the various types (Variant A to D), additional accessories will be included and must be assembled during the installation at site. In this configuration, a monitor base will be included. See the appendix for “Monitor Mounting-Plate Assembly”. Installation of the monitor base (Variant B) Remove the left upper side panel (face plate) as shown.

Attach the monitor base plate to the freezer frame (4 screws) as shown below. Replace the left side panel (face plate).

Thereafter, slide the monitor base onto the stud for the monitor support as shown.

Turn the monitor base such that the locking screw becomes visible. To secure the monitor base, use a 3 mm hex driver (included in the accessory package) and turn the locking screw counter clockwise (outward).




With the monitor base secured, slide the Touch Monitor onto it as shown. Note: Pls. consult the appendix for the Monitor / Mounting plate assembly.

The viewing angle can be set by adjusting the swivel head. The electrical wiring is described further above.

Variant C: In addition to the configuration in Variant B, this model comes with a PC support (2 pieces). Instead of the small monitor base plate with only one stud as in Variant B, this base plate is somewhat larger and is equipped with three (3) studs. The upper stud is for the monitor and the two lower ones for the PC support.

Replace the left side panel (face plate) and attach the monitor base as well as the PC supports in the same way as described above for the monitor base.

Slide the Touch Monitor onto the monitor base and place the PC onto the support as shown.




The electrical wiring is exactly the same as described in Variant B




Variant D: The installation of the base plate remains exactly the same as described in Variant B.

Thereafter, replace the left side panel (face plate).

The accessory package of this version comes with a single support for Touch Monitor and Mini-PC. Attach the support as described earlier.

Place the Touch Monitor and the PC onto the support.

The electrical wiring is exactly the same as described in Variant B. After the successful installation of the hardware, the software must be installed onto the PC. Please refer to the section on "Settings on the PC" and "Freezer Software Installation" further down in this manual. If the PC system was ordered from SY-LAB together with the freezer, the software is already installed and ready to be used.




Connecting the LN2 Hose to Freezer and Supply Vessel

The optional accessory package includes the flexible stainless steel hose and a set of wrenches. The conical metallic seal of the supply hose does not require any additional seals.

Attach the G1/4" end of the hose (small connector) to the freezer and the other end (3/4" UNF) to the supply vessel. Caution: Do not over tighten the nuts. Just tighten the nuts slightly with the wrenches. Re-tighten the nuts after a few minutes of a test run (cooling). Do not use supply tubes other than the original ones available through SY-LAB. The use of non – original tubes or adaptors will void all warranty requests. At this point, we again point out the importance of wearing protective clothing. All personnel working with the IceCube must be instructed on how to handle liquefied gases at sub-ambient temperatures. Read the safety instructions about handling LN2. Note: After a freezing run, do not immediately close the liquid valve of the supply vessel. Leave it open for another five minutes. Any Liquid Nitrogen still in the

supply hose will boil off and generate high pressure. If the liquid valve on the supply vessel is open, excess Nitrogen pressure can expand back into the supply vessel. Nevertheless, a safety valve is built into the IceCube. Any pressure in excess of about 3.5 bar will be released to atmosphere. When used in conjunction with an IceCube, the maximum working pressure of the supply vessel must not exceed 1.5 bar. At higher pressures, the operation of the IceCube's internal magnetic valve cannot be guaranteed.

Pt-100 Transport Protection

During transportation, the Pt-100 Sensors are secured in a block of Styrofoam, plastik tubing or similar material. This keeps the sensors from bouncing around. The sensors are very sensitive and should be handled with care. Never bend or drop any sensors. Do not throw away the Styrofoam block or plastic tubing. You might need it for future shipments. Note: For normal operation of the freezer, remove the transport protection from the Pt-100 sensors.

Description of the Instrument Components and Connections

Before the first turn ON of the freezer, re-check all connections. In the following, we will describe each component in more detail. These are:

• Freezer Electronic / Box • Touch Monitor • Printer (optional) • PC-System




Freezer Electronic / Box

On the Freezer Electronic / Box, you will find the main switch, power connector, fuses, the Pt-100 temperature sensor inputs and the COM serial port (9 - pin Sub D)

For replacing the main fuses, a small Philips type screwdriver will be needed. Use only the type of fuses mentioned in the section "Fuses". Caution: Before exchanging fuses or performing any internal work, always disconnect the instrument from the power line (mains).

On the topside of the freezer electronic, connections for the Pt-100 sensors (2 standard and 2 spare) and the serial control line are provided.

In the standard configuration, the 2 spare holes are fitted with blind plugs. The IceCube can be ordered with up to four sensors. A later up-grade from two to more sensors can only be done at SY-LAB. Communication is established through the 9 pin sub-D serial connector. An appropriate cable is included in the accessory package. There are four connections on the Touch Monitor (Variant B to D).

• Power Supply • VGA • Touch Monitor USB • Audio

The USB cable is hard wired to the monitor and cannot be removed.




Establish all connections. Power supply to power distribution box, VGA, Audio (not on all units) and USB cables to the corresponding inputs on the PC. For hard copies, an optional printer would be advisable. It can be any Windows compa-tible printer using either a parallel or USB connection. Now, all connection to PC and power distribution box should be established. Depending on the type of PC you are using, there might be a switched power-output on the PC. The monitor normally uses this switched power-out connector. We would like to point out, that for the operation with the IceCube, this connector must not be used.

Turn ON/OFF and Check of the System

The freezer system consists of various modules or components. It is therefore important to follow a certain turn-on and off procedure. Turn on in this order:

• Freezer electronic • Monitor • Printer if available • PC

Important: For synchronization, the monitor (especially TFT) must be turned on before the PC is switched on.

As for turn-off, it is important to shutdown the computer system in an ordinary fashion; otherwise, possible data loss will be the result. Also, you might have trouble booting up your system the next time. Please consult your PC manual for proper Shut-Off. After the PC has been shut-off, all other components can be turned off. Please note: The PC system needs some time to boot-up. Generally, the boot-up should not take longer than about one minute. Depending on the settings, the IceCube can be started automatically when entered into the AutoStart list or started manually by clicking onto the IceCube icon.

Storage After every freezing cycle, the freezing chamber should be warmed up to about ambient temperature. This can be accomplished by just starting a program, which has its start temperature at about 20°C. During the heat-up, keep the chamber closed. This has the advantage that very little condensation will occur inside the chamber. Once warmed up, dry the chamber and the lid with soft cloth or a sponge. If necessary, cleanse the system with a proper disinfectant. Caution: Before cleaning the freezing compartment, disconnect the IceCube from the power line (mains). Store the IceCube in a dry and dust free environment. Do not store below +5°and above + 40° Celsius. Do not exceed 90% humidity (non-condensing).




Handling of Temperature Sensors There are typically two temperature sensors in the freezing chamber. The sensor feed troughs are color labeled.

The red sensor is the chamber sensor, the blue one is the sample sensor. When standing in front of the IceCube, the sensor closest to the operator is the sample sensor. At all times, the sensors must be kept free of moisture. The sample sensor (blue) is used for monitoring the temperature inside a sample. IceCubes with additional sensors are available. It can be of an advantage to measure the temperature of samples at different points, e.g. in large bags. The temperature sensors are very thin, allowing us to insert them into 0.25 ml straws. In addition, the low mass of the sensor will give us a fast reaction time. When used in an ampoule, never try to force a sensor into an ampoule cap. It is necessary to pre-drill or punch (with awl, nail or small screwdriver) a little opening into cap. Prepare the opening such that the sensor can be pushed easily through the hole. Also, never try to pull out a sensor from a frozen sample. Damage will be the result. To remove a sensor from the wall-holder, pull the sensor cable slightly out of the feed through. The sensor can now easily be removed.

Caution: The chamber sensor (red) must never be inserted into a sample. This sensor controls the temperature in the chamber. If this sensor is obstructed, the chamber temperature cannot properly be controlled. Various different rack-systems are available from SY-LAB. Choose the one most suitable for you (e.g. ampoule- or straw racks, racks for homo-grafts, bone marrow or blood bags, etc.). If your system is equipped with an auto-seeding option, various racks for ampoules and straws are available too. If only small quantities of samples are frozen, the freezing chamber is nearly empty; a spacer can be used for proper positioning of your samples closer to the lid. Since the samples are located closer to the top of the chamber, it will be easier to visually check the samples (due to the see-through chamber lid) during the freezing process, especially during auto-seeding. As already mentioned, the blue sample sensor can be inserted into a sample (straw, ampoule, bag, etc.). The red chamber sensor can either remain in the wall socket or be positioned in a well-ventilated position near the sample material.




Note: The chamber sensor (red) should not be positioned in between sample materials. The heat released during the crystallization process of the sample influences the temperature reading and might alter the temperature in the chamber. Once, a program is optimized and working well, it will not be necessary, to continue inserting the sample sensor into a sample, especially if the sample quantity is limited. However, if enough sample material is available (for record keeping purposes), it will be advisable to monitor the freezing process for every freezing run performed Naturally, the sample used for monitoring must not be used on a patient. It must be discarded. In many instances, if the concentration of a sample in a cryo protectant solution is small, the cryo protectant by itself (a „dummy“) can be used for record keeping. Just remember, the sample sensor will show the temperature within the sample during the freezing process. In other words, you know what is going on inside the sample. For larger freezing bags, sensors with a length of up to 200 mm are available (standard length is 65 mm) Once more please note: For normal operation of the freezer, remove the transport protection from the Pt-100 sensors.




Freezing Chamber Description

The IceCube 14S Freezer is equipped with a transparent glass cover (lid). In this way, it is possible to visually monitor the sample material during the program run. This is especially important when determining the point of crystallization during an Autoseeding run.

Another feature is the hinge. The glass cover can be opened without looking for extra desk space. Due to the weight of the lid, the chamber is firmly sealed towards the top of the freezing chamber. An opening is provided at the chambers backside for expelling the gaseous Nitrogen. As seen from the front, there are two temperature sensors (standard configuration) positioned on the left side of the chamber wall. Straight ahead towards the right side, a plug is provided for the connection of the AutoSeeding rack. It is available as an option and can be installed at SY-LAB upon ordering or be installed in the field later on. AutoSeeding Option Kit # (14S): 13-159532 On the right side wall are the in- and outlets for the air-stream. Nitrogen gas mixed with air will be withdrawn at the top of the chamber, sweeps over the heater element (750W) to mix with the incoming Nitrogen

to be expelled at the bottom of the chamber. So the flow of the air stream is from the top to the bottom. The turbulence is generated by a fast moving fan, dispersing any incoming Liquid Nitrogen to gaseous Nitrogen.

At very low chamber temperatures, some liquid nitrogen droplets are allowed to enter the freezing chamber but will evaporate quickly off.




Available Freezing Racks Vertical Autoseeding rack for either 2 or 5 ml size ampoules.

A maximum of 36 ampoules can be placed on one rack. One side of the tubing connection is inserted into the AutoSeeding outlet at the backside of the freezing compartment. The other end of the tubing is guided through the large opening of the chamber and leads to the outside of the chamber. Caution: Make sure there are no delicate Instruments behind the Freezer. The extremely cold gas expelled at the rear exhaust might damage them. AutoSeeding rack for straws. It is available for 0.25, 0.5, 1.0 and 5.0 ml

straws. Additional straw holders are available.

Ampoule rack. It is available for 2 ml or for 5 ml ampoules. Up to 5 racks can be placed on top of each other for 2 ml ampoules and up to 3 racks for 5 ml ampoules each holding up to 96 (2ml) or 80 (5ml) ampoules.

Note: The freezing racks shown in this section undergo constant refinement and might not look the same in the future. A typical temperature sensor positioning in an ampoule is shown. The tip of the sensor should be inserted to about half of the liquid level.

It is important that the tip of the sensor is not touching the sidewalls of the containment (Ampoule). Keep the sensor centered. This can be accomplished with a centering disk made of plastic or similar material.




A rack for the vertical freezing of bags is shown below. This type of rack is adjustable and can be used for a variety of bags. Up to 7 profiles can be placed on this rack depending on the size of the bags. Size of profile: 184 x 100 mm (7,3 x 4 in.)

It comes standard with 7 profiles and allows the placement of 6 bags horizontally (depending on the size of the bag). If smaller bags are used, a higher number of bags is possible. Vertical rack for metal cassettes. Up to 11 cassettes can be place on such a rack.

For special applications, we have the possibility to produce customized racks as well.

The next picture shows such a non standard rack Rack for 20 ml glass bottles.

Need a special rack for your samples, pls. ask for a quote. Better, send us a sample of your containment (bag, ampoule, glass, beaker, straw, etc.). In the following example, we will show a standard ampoule rack resting on a spacer. This simulates a real time situation. The sample sensor is positioned in one of the containments, here in an ampoule. The chamber sensor remains in the sensor holder. Depending on the application, the chamber sensor can be positioned at any place within the chamber. In many cases, the chamber sensor is placed next to the sample material to react as close to the sample temperature conditions as possible. It is important that the chamber sensor is not touching any sample material, ampoules, freezing racks, etc. It should measure the temperature of the nearby airflow.




Various sizes are shown. The ampoule rack can be used for 2ml, 4 or 5 ml ampoules as well as for many other vials.

Please consult our accessories list at the end of this manual for more info on chamber furniture. One of our furniture is the straws rack. Because of the amount of straws that can be positioned into the IceCube14S chamber (up to 1080, 0.25ml/ 0.5ml), the handling of straws normally becomes difficult. In order to position the straws quickly onto the straw rack, we have made a device, called "Straws Distribution Block".

The Distribution Block is placed below the straw rack. Then, the straws are placed onto the rack. Because of the slits in the Distribution Block, the straws will be equally spaced. Use a small ruler to straighten out the horizontal position of the straws as shown below.

Once, the straws are positioned, the straw rack can be lifted off and be stacked up to a maximum of 30 racks / chamber on the 14S, each holding up to 36 straws.




The illustration below shows 3 empty racks stacked together.




Program Log-In Note 1: The surface of the Touch Monitors screen must only be touched with the soft tips of your fingers or with a soft stylus made out of plastic (such as the ones used on handheld PC's). Sharp and rough edges can lead to damage of the screen surface. Scratched or damaged surfaces are not covered by SY-LAB's instrument warranty. Before operating the Touch Monitor, it is mandatory to read the manufacturers instructions and warranty terms. --------------------------------------------- Note 2: Initially, the IceCube Series program does not know which of the Freezers (15M/L-14S) is connected to the PC. Only after the initial start of the IceCube Series software, the subsystem of the particular freezer reports back to the Software and will "mention" its name.

-------------------------------------------- When the IceCube program is first started, you will be asked to enter a User Name and a Password. For first-time user, the User name and Password will be in both cases

sylab Use the provided On-Line-Keyboard to enter YOUR User Name and Password if already assigned. If you do not have this information, ask your IceCube Administrator for it. The Administrator has the rights to sign you up as a new user. When using the above first-time User Name and Password, the administrator is advised to change the default User Name and Password from sylab to a real User Name and Password (this procedure will be described further down).

Here, we enter, (first time users will enter sylab in both fields – consult the section on “User Administration” to set up a new – real user) E.g. Garner Jeff and Password Press OK to get into the IceCube program or QUIT to revert back to the desktop. If you answered OK, a new screen will be presented to you like the one shown below.


Description of Program Tools and Buttons The IceCube program includes several graphical tools. In addition, Windows® based command structures are available as well. In the following section, we will describe the terminology of the various buttons, display fields, and so on.

Main Menu Pop-up menu. Behind this menu button, sub menus are found for

PlyoHesy

In geIn whtemIn elatemfurIn thePalanVe

Status Area

Display Area

Main Menu

Instrument Panel

Logo Area

Operation Panel



File, Display Options, Version information and Maintenance.

ease Note: The access to various pop-up menus depends on the USER-LEVEL granted to u by the IceCube Administrator. re, we assume that all levels (administrator level or when initially logged in with sylab -

lab) are granted.

the File menu, patient information, freezing programs, freezing runs and reports can be nerated or edited as well as the data and program path (directory) be set. Display, one can choose the way the program curve will be displayed. In Total View, the ole temperature range +40°C to -180°C can be displayed. Tracking view follows the program perature and shows a somewhat narrower temperature range.

Settings, the parameters for X-axis (time) and Y-axis (Temp.) can be defined, remaining or psed time be shown on the screen, the current program temperature be displayed next to the perature curve and the pressure curve be displayed. A more detailed description can be found

ther down in this manual. Options, some sub menus pop up. In Device settings, various alarms can be turned on/off and pressure alarm limits be set. Users can be administered, giving each user his own Name and ssword. Finaly, the language can be selected. At the moment, only German and English guage files are available. More language files will be available in the future. rsion Info gives us some information on the installed PC-program version as well as the




version number of the instruments resident program. In case of failure, this information together with the instrument model and serial number should be given to the service personnel. Maintenance allows the user to perform an Icewater calibration as well as to set the correct Communication port Status Area Display of the current instrument action like heating or cooling ON or OFF, alarm 1 / 2 status, ventilation system running, online- (CTS) and text status, etc. All statuses are displayed in form of signal lights. Is the light ON, is the corresponding function active. Display Area Graphical display of program-, chamber-, sample temperatures, and so on. During the program run, the temperature curves are displayed. Instrument Panel Numerical display of program-, chamber- and sample temperature(s) and remaining time or run time of a program. The display for the 3rd and 4th sensor is only active if these options are installed. Operation Panel Buttons for starting and stopping a program run, initialization and triggering of manual seeding, Hold and Continue button, Program Load button. to StartTemp brings the freezing chamber to the pre programmed start temperature. Seeding (option) opens an additional magnetic valve, Hold keeps the freezer suspended at the current status, Next advances the program run to the beginning of the next step and Stop stops the freezing cycle. Logo Area Above the non-removable SY-LAB logo, it is possible to add an additional logo, i.e. your own, if desired. A bitmap (.bmp) format with a size of 106 x 122 pixels should be used. In the following, we will describe in detail the various tools and buttons.




Menu Area

Main Menu - File Under the Main Menu - File, six sub menus are shown as seen on the left. The Edit description will allow us to enter sample or patient specific information. Open historical data allows us to locate, view and to some extent modify already finished freezing process data. On the Validation Software package, all modifications are recorded and are traceable In the Freezing program, new programs can be written or existing programs be edited. The next two entries regarding data and program

directories give you the possibility to select your own location (directories) where data and programs might (will) be saved. Exit will close the IceCube program.

Edit description As already mentioned, on this screen, information about a sample or patient can be entered. The first field will accommodate the Freezing Program Name. You will be able to reach this screen either by selecting Edit description just described above or it will pop-up just before the actual program run is started.




If the Edit description was selected before loading a program, no Freezing program name will be displayed. The program name will be automatically entered as soon as you load a program later on. However, information about a sample, patient, operator, etc. can be entered in the following field lines. The Last field will ask you for a sample or patient name. This field will be divided into a Last and First name area. It is important to enter some specific identification in the Last field. This will help you later on in finding a specific sample or patient in your data file directory. So Last will require some input. We would suggest entering the last (family) name of a patient or the name of a specific sample. The first 8 characters of Last will be part of an automatically generated Process record file name. The remaining characters of the name will be truncated. All other field entries except the operator name are not required but are provided to enter other useful information such as Sample or Patient ID and Memos. The Operator name will automatically be filled in and will be identical to the Login name of the currently logged in user. If more than one user is operating this instrument, they must each have a separate User Name. Pls. consult your system administrator to set you up as a new user. On the bottom of the Edit description screen, the automatically generated process file name can be seen. As you can see, the first part consists of the Last name. In addition to the Last name it will include the year, month, day, hour, minute and seconds. In this way, there can only be one specific file name for each freezing process. As an example, the file name might look like this: Entering any information into the various fields is accomplished with the pop-up keyboard.

Garner_2003-03-18_13-47-26.prc This corresponds to the file name convention

LAST_yyyy-mm-dd_hh-mm-ss.prc The extension of the file name PRC indicates that it is a freezing PRoCess file.

As already mentioned earlier, this screen will also be presented to you when you press the "to Start Temp" button after loading a program. Keep in mind that in order to save a freezing process record, we have to give it a specific name. Therefore, you cannot start an actual freezing run before giving it a Last name. You MUST enter something in the Last

field. Also, use for the Last name only characters which are allowed under the Windows® file name convention. These are all characters A-Z, a-z, 0-9. Do not use special characters such as / \ < > , . ü,ä, etc. Two Memo fields are available for adding additional information. These two fields can be edited even after a process file run whereas all other fields are protected against any changes. Just keep in mind, after a freezing run, the operator, patient name, ID, etc. must not be changed for safety reasons. However, some additional information, e.g. about the freezing run might need to be recorded (in the Memo field). However, any changes made to the in Memo field will be traceable.




On the right of each memo field, a message will let the operator know of how many changes were made to this particular memo field. If modifications were made, "Show" will list all changes that were made in the past. It will show who the operator was and at which time and date the "editing" occurred.

In our example, it will look like below. Zero "0" modification(s) has changed to one (1) showing that one modification was done to the particular memo field.

As soon as the requested information is entered, you must press OK




Open historical data For viewing freezing process records, we have to get into the Open historical data menu. Either a clear freezing process screen is displayed or a previously loaded process record is shown. On the right bottom of the screen, the Menu button gets you into various sub menus. At this point, Show description will display the patient or sample information like in Edit

description above. Show program will display the original freezing program if one was loaded with Open process record and Display Total View will show both, the program curve as well as the associated freezing run. In the File menu, you can choose among Process directory, Open last run, Set Source directory, Print Diagram, Print Program & Events, Print both of above and Close window.

Process directory When Process directory is selected, you first have to select a process record from a file list, assuming that you have made a freezing run previously. If not, this list will be empty and no historical data exists.

Open last run Open last run will display the last freezing run you have made.

Set source directory Set source directory allows you to choose the location where you want to store the Process record files.

Print Diagram, Print Program & Events, Print both Print Diagram, Print Program & Events and Print both of above will produce a printout of the process curve, process curve and a tabular listing of events ( Hold, Continue, Next, etc.) or both of the previous. Before printing starts, a preview will be shown on the screen.

Close window Close window will allow you to exit this page and to revert back to the previous screen.




Let us go back to the Process record for a more detailed discussion. If this menu was chosen and a process file opened (e.g. Testcurv_2004-07-....), following screen will be displayed.

The process record will then be displayed, can be enlarged, etc. and be printed out. The above curve is a typical freezing process test curve to find the temperature of crystallization. However, here we can see clearly the program temperature curve (green), sample temperature curve (blue) and hidden behind the green curve

the chamber temperature curve (red). The area of interest can now be zoomed in (to enlarge the display area, draw a rectangle around it from left upper to right lower corner) and be Added to the List on the right. You can choose as many windows as

you like. When added to the list, the window can be printed out later on. In the right area field, you will see the windows information.

Hier, 23.96 -4.13 44.52 -30.00 It means that the left upper corner of the selected window is at 23.96 minutes and -4.13 °C. Whereas the right lower corner of the window is at 44.52 minutes and -30.00°C.




Choosing "Events" on the above screen, additional Information about each "Action" performed during the program run can be displayed. If this is all we want to print out, press Menu - File - and choose among the various print options.

• Print Diagram: As the name implies, this will only print the diagram(s)

• Print Program and Events: Will print all the program steps and events like opening and closing the lid, pressing manual seeding, stop, etc.

• Print both of the above: Again, as the name already implies, both of the above, diagram(s), program steps and events will be displayed on screen and eventually be printed if you desire so.

If you choose Print both of the above, the result might look something like the display below.




List of Available Tools

The size of the graphical display can now be modified using

the three buttons shown.

The arrow buttons work like buttons on a tape recorder. The two middle buttons move through the various pages, page by page, whereas the two outer buttons move the display either to the first page

or the last page. Some of you might have only one page to display. In this case, the arrow buttons are not used. The printer sign on the left is used for the printer - setup. The diagrams can be printed out locally or be sent to a remote (network) printer. A message (description) appears when the cursor is positioned over an icon. Close reverts back to the main screen.

Show description, Show program, Display Total View In the sub-menu Main Menu - File - Open historical data - Menu, we have the possibility to look into the Freezing Prozess Information (Show description), look at the original program curve (Show program) and display the complete program curve and process file (Display Total View). Naturally, a process file must be opened first, using File - Prozess directory (choose your process record) or Open last run in order to display the last freezing process data.

We have now briefly discussed the Edit description and Open historical data menus. The next menu Freezing program will allow us to write a new or edit an existing program.




Freezing Program As the name already implies, here we can create a new program or edit an existing program. Actually, we are getting again in the program edit screen as seen below.

When Create new is selected, a new program with only two steps (0 and 1) will initially be displayed. Step #0 represents the start temperature. It is the temperature, at which the sample should be loaded into the freezing compartment. In many applications, this will be either 20°C for ambient temperature or about +4°C for refrigerator conditions. The default starting temperature value is +20°C.

Also, as a default, the slope in step #1 (temperature gradient) will initially be set at -1.00°C/min. In order to edit an existing program, we would choose File / Edit existing and select the program we would like to edit. More on this, further down in this manual.

Set data directory The process file of a finished run has to be saved somewhere. Here, we have the possibility to choose an appropriate location. For clarity and ease of locating your data later on, we suggest putting the data sub directory in the same location as the IceCube freezing program. Assuming that you installed the IceCube program in a directory called Freezer, a data directory called Process Records will be created. In this Process Record directory, all future process records (data) will be saved.




Note: In order to prevent data from being lost, do not forget to backup your process records periodically.

Any other directory can be used. Also a new directory can be created.

Set program directory As already mentioned above in Set data directory, the same applies here. You simply can choose the location where to store your freezing programs. Again, we suggest storing the freezing programs in a sub directory of your IceCube program directory. But again, any other directory can be used. Also a new directory can be created. Note: To avoid loss of data and programs, it is advisable to make occasional back-ups.

Exit The last file menu entry is Exit. As in most other Windows® based programs, Exit will close all processes within the program and reverts back to the desktop area. All running processes must be stopped or finished otherwise, you will not be allowed to Exit the IceCube program.




Main Menu - Display In this menu, we can set the way the program curve will be presented on the screen during a freezing run. Three sub-menus are available, Total View, Tracking View and Settings.

Total View Total View will give us an overall view of the program temperature. This is currently between + 60°C to -200°C maximum. However, if the last step in the freezing program ends at e.g. -120°C, Total View will show a screen with the lower temperature limit set at -120°C.

Tracking View Tracking View will follow the temperature curve as the program run progresses. This will give us an optimal presentation of data on the display.

Settings The remaining Menu entry is Settings. Setting allows us to define how the screen behaves during a freezing run. A freezing run may need a long time, thus requiring drawing curves beyond just one display screen. So we have to tell the program at what stage to update the screen contents for both temperature and time axis.

In addition, we can choose if we want to get the elapsed or remaining time displayed in the instrument panel. Also, we can choose to get the temperature displayed on one of the temperature curves.




Time axis On the left side of the screen, the time axis can be set. In our example, during a program run, a time window of 18 minutes will be shown. When the temperature curve approaches the right edge of the display area, just 0.5 minutes before, the screen will shift to the right by 1 minute.

Temperature axis The setting of the temperature axis functions somewhat similar. However, the up-shift as well as the downshift can be set independently. In our example, the display range is set to 40°C. When the display reaches the upper or lower edge of the display area, the up- and down shift adjusts the screen by 10°C. In this example, the up- and downshifts are set equally. Under normal operating conditions, it is not necessary to change these settings.

Time elapsed / remaining The second instrument from the right on the instrument panel is showing either the remaining or elapsed time of a program run depending on the setting above. Just mark the button show elapsed time or show remaining time as desired.

Actual program position During a program run, the current temperature of the program temperature curve can be displayed just above or below the curve. When Actual program position - Display numerical temp is marked, a little circle appears right at the tip of the program temperature curve.

Temperature value to track Since we marked - Temperature value to track -, the screen will focus on and will follow the e.g. Chamber temperature

LNi pressure LNi pressure - show curve is not implemented in the 14S. If all settings are done, press OK to revert back to the previous screen.




Main Menu - Options In the Options submenu, the Device Settings can be changed and saved, the Users be administered and the Language be selected. Device Settings and alarm configuration: To get into this menu, select Main Menu - Options - Device Settings. Following screen will be displayed. In all cases, you can see that besides an internal alarm, additional alarm outputs can be set. However, this requires additional hardware within the freezer. The additional alarm outputs can be ordered as an option with the freezer directly or be upgraded later on (freezer box must be sent to SY-LAB). The alarm-relay outputs provide potential free contacts rated at 250VAC / 10A.

Here, we can set the condition and duration of various alarm outputs on the IceCube Freezer. In short, these are:

• Indication that start temperature has been reached • Alarm outputs when: • temperature deviation is out of the set limits • defect of temperature sensor • a critical power failure occurred • a short time power failure occurred • the freezing program is finished

Note: LNi pressure Alarm not activated on 14S units




Description of the various conditions and alarms Note: All displayed values are the default settings

Start Temperature reached condition On the right, you can see the corresponding display-field. Here we can determine how a start temperature reached condition is defined. In our example, if the start temperature of the freezing chamber (defined in the freezing program) is within +/- 0.5°C and if this condition prolongs for at least 20 seconds, the start temperature reached indicator will light up. As you know from the above description, the "to Start Temp" button will change to "Start" when the program is started. But when the program reaches the start temperature and meets the conditions shown above, the "Start" buttons will be illuminated. This functions only once. As an example. If the start temperature is reached, the button lights up. If however, START is pressed, the light is turned off. If the cover is now opened, the temperature now rises above the start temperature and returns back after the cover is closed, the Start button will not be illuminated again.

LNi pressure Alarm (not implemented on 14S) During a freezing run, the pressure in the supply vessel will be monitored, thus giving us some information on how a freezing program behaves when the pressure changes. Here in our example, an internal alarm (Buzzer) will be triggered when the pressure falls below 0,1 bar or when the pressure exceeds 1,5 bars. Besides getting a corresponding message on the screen (when the PC is operational), an acoustic alarm will be heared.

The appearance of this acoustic alarm depends on the settings of the four time fields. In our example, we turn the buzzer ON for 1 second, and then we turn it OFF for 1 second. Again we turn the buzzer ON for 1 second but now, we turn it OFF for two seconds. So this alarm will have a distinct sound, like

Beep (1sec.) - Break (1 sec.) - Beep (1 sec.) - Break (2 sec.)

Note: In the following alarm descriptions, the internal buzzer alarms will behave in the same way as just described and will not be mentioned anymore.




Allowed Temperature deviation Whenever the actual chamber temperature deviates from the program temperature by more than the pre-set value, an acoustic alarm will be issued according to the settings on the right. Here, the internal buzzer will give an alarm whenever the program- and chamber temperature deviate more than +/- 5°C. If the Temperature deviation is getting again within the specified limits, the Alarm will be resetted.

Temperature reading invalid The readings of the temperature sensors are always compared with values in a tabular listing. If the readings are wrong or out of some limits due to a sensor short or break, a visual as well as an acoustic alarm will be issued according to the settings on the right.

Critical power fail - awaiting user assistance As already mentioned earlier, a critical temperature window can be defined. This will be most likely the temperature range in which crystallization of the sample takes place. In case, a power failure occurs within this critical temperature window, the alarm conditions on the right apply. In addition, you will get a corresponding warning message on the screen, when power is re-established to the PC.

The warning message also will list the time at which the power failure occurred. It is now important to determine in which state the sample is. According to the sample condition (frozen or not), the operator has to take an appropriate action. Either to discard the sample, re-run the freezing procedure or placing the sample into Liquid Nitrogen

Also, another window will be displayed. The Re-Start Point window. Here, the user has now the possibility to determine where the program should continue with the program run. Either time or temperature can be selected. It is also possible to set the re-start point graphically by positioning the cursor at the point of restart.

Short time power fail A short time power fail is a non critical alarm. Since the power failure occurred only for a few seconds, we can assume that this did not harm the sample. As soon as power is re-established to the freezer, the freezing program will




proceed as if nothing would have happened. Some hospital power supplies will take a few seconds to switch over to the auxiliary supply. So the IceCube Series Freezer can handle such systems and continues to work. Nevertheless, an alarm condition is issued, acoustically as well as visually. The visual warning message will be seen only when power is re-established to the PC.

Freezing program finished At the end of a program run, most operators would like to get notified (e.g. to transfer the sample(s) to Liquid Nitrogen). For this purpose, we are using the program finished alarm. It will give an alarm but can be set such as to give alarm on the internal freezer buzzer as well as on the local PC.

Save Settings To keep those changes also for the future, it is necessary to Save the Settings in memory.




User Administration One of the menus in the Options menu is "User administration". User administration allows the creation of a list of users. Each user has to have a specific name and password in order to Log-In into the IceCube Series program. Depending on the user rights, one has access to functions that others do not. Initially, IceCube Series software is delivered with the administrator log-in name and password set to sylab. We strongly recommend to change these settings top your personal User Name and Password. Note: Before deleting sylab, make sure you have tested your personal log-in. The following screen opens if logged in as an administrator with all rights

or this screen,

when logged in as normal user, having no administrator rights.

Note: Again, depending on the settings of the user rights, the user will have only access to functions for which the administrator gave permission to use them. Initially, when Logging-In as sylab , administrator rights are granted. The administrator should then create his own User Name and Password, create for himself administrator rights and set up other users with limited or equal rights. The user sylab should then be deleted.




First, you will be presented with a list of users if they were previously entered. To create a new user, select New user. The keyboard pops-up and you will be able to enter a user name, password and password verification. Each field needs at least 5 characters. Just press OK to accept the new "USER". If you have not entered exactly the same characters in the password verification field as you have entered in the password field, an error message will be displayed

"password and verification do not match" Correct the problem by typing in the same information in both fields. Also, if the user name or the password is too short, it will let you know "minimum required length is 5 characters". In addition to enter a new user, users can be deleted or passwords be changed. Once you have entered a new user, Close form to revert back to the main screen.




Info for the Administrator Administrators have the right to grant or not to grant other users access to various functions. Following functions can be selected or de-selected. These are: Look at previous freezing runs----------------------------- All available programs can be used ---------------------- Start the act. prog. before the start temp is established Stop freezing run before the program is at the end ---- Allow the user to use the HOLD/CONT. button------- User has permission to jump to the next step---------- Give permission to turn on manual seeding-------------- Not available on 14S (Trigger) ----------------------------- Allow the user to write or edit a freezing program ------- Permit the modification of the two memo fields --------- Change the path of the data directory ---------------------- Change the path of the freezer program directory ------- Choose another language ------------------------------------ When changes are made, allow to save them perm. ---- Allow the user to make changes to the device settings - Let user perform an ice-water calibration ----------------- Give permission do change load device settings --------- Allow the user to update the subsystem of the frezzer - Give permission to use a different communication port Grant administrator rights -----------------------------------




Language In the IceCube Series software, you will be able to select among one or more different languages. At the moment, only German and English are available. Other languages can be added if a need for it arises. The language file name ending will be *.LNG. For example: Engl117b_1.LNG

When you open the language menu, you will be asked to choose the directory location and the name of the language file. Please note: Once you have selected a different language file, it is necessary to close and re-open the IceCube 14S freezer program.

Using the new language file as the default, it will be necessary to press "Save Settings". Next time the freezer is turned on, it will have the new language file setting.




Main Menu - Version Info Behind this menu, we can find the program version of the currently installed IceCube program. This information together with the serial number of the instrument will allow us to search through our production records. It is like the fingerprint of your instrument.

Main Menu - Maintenance There are two submenus in Maintenance, Ice water calibration and communication.

IceWater Calibration From time to time, it might become necessary to perform an ice-water calibration. Electrical components are slightly changing their characteristics with time. So it is advisable to occasionally re-calibrate the temperature sensors. In any case, if a temperature sensor is exchanged, an ice water calibration must be performed. Get some crushed ice and pour it into a mid-size coffee cup. Add some tap- or distilled water to it. Stir thoroughly to get a good mixture of water and ice (slurry). Now take the temperature sensor(s) out of the chamber holder and place it into the slurry. During the calibration, slowly agitate the sensor(s), making sure that the temperature distribution in the cup is constant. Do not touch the walls of the cup. Also, hold the sensors as shown in the picture in order not to influence the temperature readings. Agitate for a minute or so to reach thermal equilibrium. Enter the Ice Water Calibration screen by selecting Main Menu - Maintenance - Ice water calibration.




The calibration screen opens, showing you a lot of temperature readings and digital data. Don't worry, it is not complicated and you do not have to enter or change any of the values you see in the table. The only functions we can use on this screen are:

Calibrate Load from ....

Save as .... upload

When you receive the instrument from SY-LAB, you also will get a diskette with calibration data. These are the default calibration parameters for YOUR freezer. They are stored in the freezers memory

already and are provided only for the purpose of having the default parameters available in case somebody overwrites the internal calibration parameters with erratic data (e.g. performing an ice water calibration at ambient temperature). If the calibration parameters are lost, they can be reloaded, using the function Load from. When the freezer undergoes a new ice water calibration, the new calibration data should be saved. This is accomplished by the Save as ... function. The actual calibration is performed with Calibrate. Now, let us continuo our discussion from before. With the particular sensor in the ice water slurry, press the corresponding correction button at the bottom of the screen. The first field is for the Chamber temperature sensor, the second field for the first Sample temperature sensor (other temperature sensors are optional) and so on. When you feel that the slurry is agitated well, press the Calibrate button. Chamber Sensor Sample Sensor Additional Sample Sensors

The reading from the temperature sensor at 0°C is then compared with the tabular listing. The difference or the correction value will then be displayed in the corresponding field in °C. After calibration, the new calibration data must be uploaded to the electronics. Press upload. Also, store (Save as ) the new calibration data either on Disk or write it to a sub -directory for later retrieval if necessary.




Communication In the communication menu, we can set, as the name already implies the communication port (COM-Ports). Depending on the number of COM-Ports on your computer, you will be able to select among COM1 / COM2 / / COMn. Select the correct COM port and "Save Settings".

At this point, we will continue with writing a little test program to get you started.




Writing a Test Program Note: The following description assumes that full administrator rights are granted.

Test program for finding the crystallization temperature of a sample

In this section, we will describe in detail how to write a new program. At first, we will use a typical test program for finding the crystallization point of a sample. Keep in mind that for every individual sample material, a new program curve must be found. In our example, a 2ml ampoule, filled with 1ml of a 10% Glycerol / Water solution will be used. Since this will be a fairly easy program, it is perfect for getting the hands on the equipment. Following the test curve programming, we will use the results from the test curve to create an actual freezing program curve. As already mentioned, we will try to find the crystallization temperature of a 10% Glycerol / Water solution. For this purpose, we slide the tip of the sample temperature sensor through the cap of the 2ml ampoule. Getting good temperature distribution results requires the tip of the sensor to be in the geometrical center of the solution. The sample temperature sensor (blue) is the one closest to the front of the freezer. Now we can start creating our first (test) program. For this purpose, we will focus on already existing programs. The start temperature of the freezing process depends in most cases on the initial temperature of the sample or freezing (cryo) protector. Is the cryo protector at refrigerator temperature - (about 4-5°C) and the sample + cryo protector kept at this temperature, then, our freezing process should start at this temperature. However, is our sample / cryo protector at ambient temperature, then we will start the freezing process at ambient temperature - about +20°C. In our example above, we will start the freezing process at +20°C. Then, we will bring the cryo solution (sample + cryo protector) down to zero (0°C). At zero degrees, we will remain there for some time to give the cryo solution time to equilibrate at this temperature. In other words, we give the cryo solution time to get close to the programmed temperature. In this way, we always start with the same sample temperature conditions. So we will cool the cryo solution with -2°C/min. from +20°C down to 0°C. Then, we will keep the temperature at zero for 5 minutes. Please note: This is only an example. Cooling rates, equilibration time, crystallization, and so on might be different for your sample material, cryo solution, etc. The point of crystallization should be found while slowly cooling the cryo solution. As slower the cooling takes place, the higher the chance that we find the exact crystallization temperature. However, if the cooling (slope) is too slow, the program will have to run for a very long time. This in turn will lead to a high Liquid Nitrogen consumption. So we have to find a compromise. A cooling rate of -1°C/min. is widely accepted and used in many applications for approaching the crystallization temperature. From experience, we will assume that the crystallization temperature of our Glycerol / Water solution will be somewhere around -7°C to -10°C. So in order to cover this temperature range, we will program a lower temperature of -40°C. In other




words, we will have a temperature step going from 0°C with -1°C/min. down to -40°C. This is all we need for our test curve. In summary, the program, let's call it TESTPROGRAM will look like this:

Step Temp - °C Slope - °C/min. Time - min. 0 20,00 0,00 1 0,00 -2,00 10,00 2 0,00 0,00 5,00 3 -40,00 -1,00 40,00

With the information above, we will now program our first freezing process. For this purpose, we have to get into the Main-menu - File and select Freezing program - Create new.

Freezing program editor

Edit control - Program The following screen will be opened.

Edit Freezing program points

Initially, one temperature segment (step) - +20°C to +10°C, with a slope of -1°C/min. is displayed as a default. At first, the program segment will be displayed as a green line. We have now various possibilities to enter our program. One would be to graphically move

the end points of a temperature segment around. This is OK for getting an approximate program curve.




Another method would be to use the arrows next to the temperature, slope and time fields. The various steps are then shown in a tabular listing at the upper right of the screen. However, entering an exact program will require the input through the keypad. In this way, all entries for Temperature, Slope and Time can be entered exactly as we want them. To open the keypad, just click in the Temperature, Slope or Time field. Immediately, the keypad display opens. A second double click in one of the fields will close the keypad again. The Temperature, Slope and Time values can now be changed numerically by typing in a value or values can be changed by using the up and down arrow buttons. The left arrow is used as a back-step button (removes character by character to the left of an entry). In addition, there is a delete (del), clear input, enter and minus (-) button.

Now we want to keep the first temperature point at +20°C. The end-temperature point

however should be at 0°C with a slope setting of -2°C/min. Touch the green end point of the first segment if it is still green. The green line will turn to red. Now, double click in the temperature field. The keypad opens. Enter 0,00 and press Enter. The value in the temperature field will change to 0,00 and the Slope field will be highlighted next. Type in -2,00 for the Slope and press Enter. The Time will be adjusted accordingly. Here, it will display 10 minutes. Naturally, a temperature difference from +20°C to 0°C with a speed of -2.00°C/min. will take 10 minutes. Let us enter the next step in the program. Press Append Point. The previous segment (step) will turn green and the new step will be red. Repeat the procedure above. This time, we would like to

remain at 0°C for 5 minutes. So we set the temperature to 0.00°C. Click into the temperature field and enter 0.00 on the keypad. Then, click on

time, press clear input and then type in 5.00 and press enter. Again, to add the last step, press Append point, enter -40°C for temperature and -1.00°C/min. for slope.




The display should look exactly like the one above. If you have made any mistakes and you want to change any of the steps above, use the Previous, Next, Delete point or Insert point buttons to correct them.

Previous will move you back in the program. Once the first step is reached, the Previous button will be disabled.

Next will advance step by step to the right until the end of the program is reached. On the last step of the program, Next will be disabled.

Delete point will remove the red (current) program segment from the screen and also from the program list on the right. Caution: If you have pressed Delete point by

mistake, you have to re-insert this program segment.

Insert point will add a program segment just before the current step (red). If step 3 is the current step, Insert point will add a step just between the second and third

program segment.

Append point will add a new program segments to the end of a previous segment. So Append will add new steps to your program.

Now it is time to save our little test program. Filing operations

The four buttons on the right are used for filing operations. File/New will clear the screen and you are ready to enter a new program as described above. File/Save will allow you to save your freezing process curve (program). You will be asked for a file location and file name if it is a new program. If you opened an already existing program, file save will re-save your file under the same name

without asking for another name.




File/Open will allow you to open an already existing program. It can then be edited and saved under the same or a different name. File/ save as will give you the opportunity to save a new or existing program under a different file name. File/ Exit closes this screen and reverts back to the Main-Screen Press File/Save and enter TESTPROGRAM as file name. If this name already exists, you will be asked to either enter another name or to overwrite the existing name.

Axis standard

Axis standard will always display a default screen width of 180 minutes and a screen height of 170°C (+30°C to - 140°C).

Mouse button

In order to provide the function of a right mouse button, one can press on the standard mouse symbol. It will change its appearance and can now be used once as a right mouse. If this function is needed repeatedly, just press on the mouse symbol.

Active point size

Active point size - will allow you to change the size and sensitivity area of the movable pointer (red dot). The larger pointer can be moved around more

easily on the touch screen and is provided only for your convenience.




Before we start the freezer to determine the crystallization temperature of our sample, let us discuss the way windows or screens are zoomed in and out, screens moved around and so on.

Moving Screens and Zoom Functions

Program curves can be very long and might not fit on one display screen. In order to move from screen to screen, a mouse button was installed on the program screen. When the Touch Monitor is touched, the function of the left mouse button is activated. In order to enlarge an area of your screen, you can draw a square around this area by moving from the left upper point to the right lower point just as you would with a mouse in a Windows® based environment To repeat, an enlargement can be accomplished by drawing a rectangle starting at the top left down to the bottom right. If we reverse this procedure, starting to draw a rectangle from the lower right corner to the top left corner, we have reset the screen to the Total View. Try it out. Do not hesitate, move your fingers over the screen and test a little bit. Now as already mentioned, this would be the function of the left button on a standard mouse. However, for some features, we will need also the function of the right mouse button. No problem. Just press the mouse button on the lower right of the screen. The mouse image will change (red right button) and will be active for only one mouse action. Thereafter, the image changes again back to the left mouse button. When will the right mouse button be used? Whenever you want to move the whole screen around! If the right mouse button is active, touch the monitor and move your finger around on the screen. You will see that the screen is shifted to whatever direction you move your finger to. In this way, it is possible to move around on the screen for optimum display comfort.




Running the Test Program In the previous section, we have learned how to write a simple program. Actually, this program can be used for finding the temperature of crystallization of our or many other samples solutions. Let us open the previously saved TESTPROGRAM. Press the Load program button and select "TESTPROGRAM" from the displayed program name list. The program will be loaded and will look like the one below. Note: Load program is only available when the IceCube has established communication with the PC. Only then is Load program visible.

Our test-program is fairly small, so it will fit only on one display screen. Following are the various program steps.




Step 1 lowers the temperature in the chamber from 20.00°C to 0.00°C with a slope of -2.00°C/min. Step 2 holds the chambertemperature at 0.00°C for 5.00 minutes. Thereafter, step 3 lowers again the chamber temperature from 0.00°C to -40.00°C with a slope of -1.00°C/min. We assume that the crystallization temperature will be around -7 to -10°C. So we have covered with this program the temperature range of interest. At the moment, we see only the program temperature. It is the green curve or line. When the program is running, we also see a red and blue curve. The red curve represents the chamber temperature, whereas the blue one represents the sample temperature (if inserted into a sample). The sample sensor is only monitoring the temperature behavior in the sample and does not take part in the temperature regulation of the chamber. However, for other functions like Autoseeding, the sample sensor can be selected to trigger Autoseeding. More on this subject in section "Autoseeding". Before the start of the program, only the Chamber- and Sample Temperatures are displayed. As soon as the program run is started, the Program temperature and the Time display will be shown as well. Sensor 3 temp. and Sensor 4 temp. will be active only when additional sensors are installed. In the standard configuration, the IceCube comes only with two sensors - Chamber and Sample sensor. Inspect the level of Liquid Nitrogen in your supply vessel. Also check that the vessel is pressurized and the liquid valve is opened. This freezing cycle might need anywhere from 5 to 10 liters of Liquid Nitrogen. Note: Use only supply vessels which are approved and confirm to the local regulations about those pressure vessels. Only trained personnel is allowed to operate such vessels. All necessary

Step 1 Step 2

Step 3




and required safety precautions must be followed. If in thought, please contact your safety advicer. You can see that the "to Start Temp" has changed its color to blue. A sign, that it is ready to be pressed. If all conditions are right, press the "to Start Temp" button. As soon as the "to Start Temp" is pressed, the IceCube will try to reach the start temperature. At the same time, the button "to Start Temp" will change to "Start".

At this point, it is possible to start with the actual program run. However, in most cases, we want to wait until the freezing chamber reaches the start temperature. When the start temperature is reached, the Start button will be "illuminated". This is the time to insert your samples into the freezing chamber. Please note also the "lamps" in the Status Area. The Fan light as well as the Online (CTS) light should be ON, telling us that a) the fan is running and b) communication (PC to Freezer) is established. In addition, depending on the initial temperature of the freezing chamber, the Cool and / or Heat lamps should be turned ON and OFF in a periodic manner.




As seen on the following screen, we have reached the start temperature, the "to Start Temp" button has changed to Start and is illuminated. Insert your sample solution / material, etc. and press Start. Immediately, the IceCube starts to run the freezing process. The screen changes and displays three curves. The green one represents the program-, the red one the chamber- and the blue one the sample temperature. Occasionally, if all temperatures are nearly the same, the three curves will overlap and only one curve might be seen. If you have activated Actual Program position - show and display numerical temp. in the Display Settings menu, the program temperature will be shown in a little window next to the temperature curve. The freezing process will now proceed. As soon as Start has been pressed, the Start button changes back to "to Start Temp" and will be disabled. Other buttons however will be activated.

As we can see in the next screen, the three temperature curves are clearly visible. The chamber (red) temperature is exactly following the program (green) temperature. The sample (blue) temperature is lagging behind, because the sample needs more time to cool. We are now waiting for the moment at which the sample temperature will be rising very rapidly. This will be our crystallization temperature of this sample.

As it happens, crystallization took place at a chamber temperature of 11.9°C. At the same time, the sample temperature was at -10.3°C. It is the chamber temperature that we are interested in. In addition, we will further investigate the sample to see how long it will remain at the elevated temperature. Small samples

will continue to cool faster than larger samples. Actually, what you see below the blue curve is the amount of latent HEAT. It will be our task to modify the test program in such a way as to compensate for this amount of heat. Ideally, the blue curve should remain parallel to the green / red curve. However, this will not be possible. The phase transition from liquid to ice will always be seen. It is on us to find a curve that will minimize the increase of heat.

This procedure is called "optimization". As we can see below, the sample curve tries to reach the program curve again. This is then the case when all the liquid has changed to solid (ice).




As long as there exists a mixture of liquid and ice, the temperature of the sample remains nearly constant. For now, we have gathered enough information to modify our test curve. Therefore, we will now stop the freezing process. Press Stop. You will be asked if you really want to stop? Press YES. As soon as you say YES, the freezing process will be stopped and the total freezing process curve will be displayed.

The freezing process has been saved to disk automatically. There is no need for additional saving. Just remember, the freezing process file will be found in YOUR specified data directory location under the name convention (e.g.):

LAST_yyyy-mm-dd_hh-mm-ss.prc

Operation Panel

Seeding Seeding - Manual - (works only if optional seeding unit is installed): Seeding is another way of freezing a sample. Instead of using a heat compensation curve (will be shown later when optimizing a curve), the main task here is to uniformly grow ice crystals around cells, embryos, etc. Please consult the section on "Seeding". A program can be written such as to perform seeding automatically. Please read the section on "Autoseeding". For seeding, optional equipment is required.

Hold Hold: During a freezing process, the operator has the possibility to manually suspend the run. This is accomplished with the Hold function. When Hold is pressed, the current temperature will be held constant for as long as the Continue button is pressed. The Hold button will be toggled to Continue after it is pressed and vice versa. In other words, a Hold / Continue operation adds time to the total length of the program. One application would be that the operator needs more time to fill up the freezing chamber. So at start temperature, he just presses Hold. When the chamber is loaded, he Continues with the freezing run.

Next Next: The Next function performs the task of jumping to the beginning of the next step before the previous step was finished. For example, if we are in the middle of program segment (step) 1




and we would like to proceed rapidly to the beginning of step 2, we just hit Next. In the section "Optimizing a Program Curve", we will discuss the usefulness of this function. Next will not work during a Hold.

Load program/Stop Load program button shows up first to load an existing program. Once a program is selected (loaded), this button changes to "Stop". Stop: As the name already implies, Stop will stop the freezing run. But before this happens, you will be asked to acknowledge this action.

You can then choose YES or NO. The process file will be saved under the following file name convention.

Sample- or Patient name_Date_Time.PRC e.g.

Huber_2003-03-18_13-47-26.prc

(max. 8 characters)_year-month-day_hour-minutes-seconds.prc Also, you will be asked if a standard report should be printed.




Writing the final program - Optimization (as an example) In this section, we are going to modify our test program such as to compensate for the heat of crystallization. In addition, we will add the remaining steps to bring the sample down to a temperature of about -120°C. At this temperature, most of the samples are then transferred to a long-term storage vessel. Let us recall what we have so far.

Step Temp - °C Slope - °C/min. Time - min. 0 20,00 0,00 1 0,00 -2,00 10,00 2 0,00 0,00 5,00 3 -40,00 -1,00 40,00

We also learned that the crystallization of the sample takes place at a sample temperature of -10.3°C. The corresponding chamber temperature was at -11.9°C. Since we want to compensate the increase of heat in the sample, we have to rapidly lower the temperature in the chamber just an instant before the warming starts. So we will modify step 3 above to cool down rapidly from -11.4°C (about 0.5°C less as the chamber temperature at crystallization) to -40°C with a speed (slope) of -60°C/min. From past experiences, it was found that compensation temperatures of -40°C or -50°C are suitable for many applications. At -40°C, we will remain at this temperature for some period of time. Again, from experience, for ampoules we will keep the temperature at -40°C for 0.5 minutes. Then, we will increase the temperature again to about -25°C with a slope of +10°C/min. To slow down the heating process, we add another step with a smaller slope. From -25°C we go to -17°C with +3°C/min. Then, we continue to cool with a slope of -1°C/min. down to -30°C. At this point, the sample(s) should be all frozen. Now, we can speed up the cooling process. From -30°C to -70°C we cool at a rate of -5°C/min. Thereafter, we approach our final end temperature of -120°C with a cooling rate of -8°C/min. In tabular form, it looks like this:

Step Temp - °C Slope - °C/min. Time - min. 0 +20,00 0,00 1 0,00 -2,00 10,00 2 0,00 0,00 5,00 3 -11,4 -1,00 11,40 4 -40,00 -60,00 0,48 5 -40,00 0,00 0,50 6 -25,00 +10,00 1,50 7 -17,00 +3,00 2,67 8 -30,00 -1,00 13,00 9 -70,00 -5,00 8,00 10 -120,00 -8,00 6,25

Use the same procedure as described above in the section "Writing a Test Program" to enter these program steps. Call this program "FullProgram".




The result will be shown below. In the display area, we see the graphical representation of the freezing process curve. On the right, we have the corresponding tabular listing. It should be identical to the table we have worked out before.

If not yet done, "save as" the new program under the name "FullProgram".

Prepare a new sample of the same material and quantity. Use the same type of ampoule. Ampoules from another manufacturer might have different heat transfer coefficients. So please keep all parameters the same. Also, do not use a pressure vessel with a different pressure. It will definitely change slightly the temperature point at which crystallization takes place.




Just remember, this is our first real freezing process curve. In most cases, this curve has to be modified repeatedly to get the optimum freezing process for our sample. According to the results of this freezing run, we will have to further adjust (optimize) this program curve. Check again if:

• the pressure in the supply vessel is adequate, • the liquid valve is open • new sample is on the sample sensor • the chamber is at about ambient temperature • the program "FullProgram" is loaded • the chamber lid is closed.

Press "to Start Temp".

You will now be asked to fill in some sample or patient information. It is mandatory to enter at least a Sample / Patient name. It uses this info for creating a unique file name. Enter "Test" for Last name. As a first step, the chamber will be brought rapidly to the start temperature. In our example, this is +20°C. As soon as the start temperature is reached, the blue Start button will light up, signaling that the freezing chamber has reached the start temperature and that the sample can be loaded into the chamber. In the screen below, we have on purpose, forgotten to press the Start button. Nothing much really happened. The chamber temperature will remain at the start temperature as long as we do not press Start or Stop or the Liquid Nitrogen runs short. Here we can see how constant the start temperature remains. When Total View is selected, the freezing process file (green) will also be displayed.




The system has stabilized at +20°C. Press Start to start the freezing process. It will now take about 26 minutes before we reach the crystallization temperature again. Normally, it takes a few runs to get a good freezing process curve. Depending on the sample material and its containment, optimizing a curve can be simple and quick or just be the opposite. We will see in about 26 minutes. If we selected in Main Menu - Display - Settings "Actual program position" - display numerical temp., we will also see the numerical display of the program temperature during the freezing run on the screen.

While this program is running, let us look at Autoseeding.




Special Function - Autoseeding As already mentioned before, there are some special functions built into the IceCube program. One is the Seeding. Seeding can be programmed so that it is initialized automatically. We can find this function in the Main Menu - File - Freezing program - Edit existing or Create new. The function "Trigger" is not available on the 14S Freezer.

Setting the Autoseeding Level At the beginning of this manual, we have followed the Program path. Now, we will look into the Autoseeding path. Click on Autoseeding. Following screen will appears

For IceCubes without the Autoseeding option, Autoseeding should be disabled. If Autoseeding is installed, the operator has two possibilities to initialize the seeding.

• by temperature of any sensor or

• by time If temperature is chosen as the seeding criteria, we have to tell the system at what temperature to start and on which Sensor to measure. Any temperature sensor can be selected. In general, the sample sensor is used for monitoring. As an example, above, we have found crystallization talking place at a sample temperature of 10.9°C. This was measured with the sample sensor in the sample (ampoule). Seeding should take place prior to the self-crystallization. So we choose a temperature somewhat less than 10.9°C. For a first try, we should enter 10.4°C in the temperature field and select the Chamber sensor as measuring criteria. Press Enter. As soon as the Enter button is pressed, a horizontal line on the graphics display will indicate when Autoseeding will be turned on. In addition, we have to tell the system for how long the seeding valve should open - Active time. Past experience has shown that seeding should not last longer than about 30 seconds. If it lasts longer, the freezing behavior of the sample might change. Note: In order to understand what seeding is doing, please refer to the section on "Seeding". If crystallization of the sample is not initialized within 30 seconds, the sample temperature is still too high (too warm). If the crystallization temperature was determined accurately, seeding times of 10 seconds are not uncommon. The shorter the seeding time, the better for the sample.




The second way to initialize seeding is on Time. Basically, if we run a freezing process, the crystallization temperature will be reached in a certain time. So this criterion can be taken as well. If we know that it takes 26.4 minutes for the sample to reach 10.4°C, we can enter in the time field 26.4 minutes. The valve open time should be the same as above. No more than 30 seconds. The shorter, the better. Now, let's come back to our resulting freezing curve.

As we can see, the sample temperature is still rising around the crystallization temperature. So apparently, the quick temperature compensation at -11,4°C was initialized too late. So we have to modify our current freezing program "FullProgram".

Step number 3 has to be changed from -11,4°C to let's say 10,3°C as seen in the tabular listing on the left. Save the newly modified curve under the name "FullProgram2". Again, we have to re-run the sample. As already mentioned earlier, always start at the same conditions. For instance, freezing chamber at 20°C, sample at 20°C, new sample of same material and quantity as in the previous run, sample sensor inserted into the geometric center of the sample, same pressure vessel, etc. The result of this curve will let us know if we have to continue to optimize the curve or if it is already acceptable. Normally, we would

have to go through another 2 to 3 runs before the curve "might" be optimized. Without documenting here the same curves over and over again, it was found that when running the program on the left, the sample temperature still increased at the crystallization temperature. So therefore, Step 3 was changed to -9.5°C and the freezing process started again. It will take about another 26 minutes before we can see the results.

We can see that even at -9.5°C, there is still an increase at the sample temperature around crystallization. So we will once more, modify Step 3 down from -9.5°C to -8.5°C and see what happens. If we compare the current freezing process with the initial one, we will discover that the area under the sample (blue) curve has become smaller. This in turn tells us that the low temperature in the chamber has compensated some of the heat given off by the sample during the crystallization.




Having now changed step 3 to -8.0°C, we rerun the sample again, assuming the system is back at starting condition. The result of this run is visible. As you can see, the increase of heat in the sample is already smaller than it was ever before. Also, the temperature increase of the sample is already somewhat compensated by the lower temperature in the chamber. However, we still have to compensate a little earlier. This time, we set step 3 to -6.5°C and see what happens.

Once we know the characteristic behavior of our material, we might be able to speed up the optimization of our freezing processes. Initially, the crystallization temperature of the 10% Glycerol / Water solution was found to be at a chamber temperature of -10.9°C. Now we are down at about -6.5°C. Naturally, the rapid cooling will somehow change the cooling behavior of our sample. Since different sample materials are behaving differently, we have to follow this procedure by trial and error. Now, the curve on the left looks again better than the previous one but still is not optimized. So we try once more to lower the rapid cooling point (step 3) to -5.5°C. At the same time, we will slightly change step 7. As it seems, once the sample temperature increase ceases, the heating time or speed of step 7 (+3°C/min) takes too long. So we increase the speed of heating to 5°C/min. Here, we see that with step 3 set to -5.5°C, the rapid increase of the sample temperature starts to flatten out. This is already a good sign that we are very close to our goal - getting a curve that is as flat as possible. So again, once more we increase step 3 to -4.5°C. Our tabular listing now looks like this: Now, let us run the sample one more time and see what happens. The final freezing process curve is shown below. As you can see now on the enlargement, the increase of temperature in the sample has almost ceased. As a comparison, look back at our initial freezing process and compare the area below the sample curve. Can you see the big difference? We just have optimized our freezing process curve.




If you have enough time, you can play some more with the temperature profile. Perhaps you get the curve even flatter. Below, the same curve as above is shown but only the scaling factor has changed. Again, we can see that the increase of heat in the sample is minimized and fairly well compensated by the temperature profile below. We are now at the end of describing how to optimize a temperature profile. It is now up to you to either accept the program curve as is or to play some more to make the curve even flatter.




Setting the Restart Level (only if no additional emergency power source / UPS is connected) Another function in the Edit control window is Restart. After a power failure, the IceCube freezer tries to reestablish the temperature conditions it had before the power failure occurred, depending on the settings in Restart.

First, a critical temperature and time range can be set. In other words, we can specify an area where our sample is most damageable. Naturally, this will be around the point of crystallization. So we will set the limits accordingly to our freezing process curve. Both criteria have to be met; the temperature in the sample after power return has to be within the temperature and time limit settings. In our example below, it will look like this.

The temperature limits are set approximately between 2,5°C and -24°C. The time limits are set approx. between 15.6 and 40 minutes. Let us analyze these settings. Case 1: Above or around zero (0°C), the sample is not frozen yet. Also, we have not reached 25 minutes yet. After power comes back, the IceCube will continue automatically with the freezing process. Case 2: The sample temperature is below -24°C and the program run time is over 40 minutes. The sample is practically frozen and the IceCube will automatically continue with the freezing process once power is reestablished to the instrument. Case 3: The sample temperature at the time of power return is within the upper and lower limit (between 2.5 and -24°C) and the power failure occurred within the left and right time limit. The freezing process will not automatically continue. In this case, an alarm will be issued, signaling to the operator to take action. The responsibility is now on the operator to decide what to do. If the sample has not started to crystallize yet, it might be possible to continue with the program run. If the sample is nearly at the end of crystallization, it might be possible to continue with the program run. However, if the sample is just at this point where the heat of crystallization is removed, your sample might be spoiled. For important sample material, we always recommend using an Uninterruptible Power Supply (UPS). Also, we can set the Maximum non-critical power fail time to bridge the time required for emergency power supplies to come up. Most hospitals will have such emergency power supplies. If not continuously operated, it might take a few seconds to start such systems. If power is interrupted, the IceCube system will stop working. However, if this interruption is only of short duration (a few seconds), the IceCube will automatically continue with the freezing process as if nothing would have happened.




The internal microprocessor of the IceCube stores all of the settings and temperature readings prior to the power failure and can therefore immediately (within 1 sec.) continue with the program run after the power is restored to the instrument. The external PC will need some time to boot up again and it might take a minute or two to see the actual freezer program on the screen again. During all this time, the freezer operates properly in the background. In addition, no data will be lost. It is up to the operator to find out how long the emergency power supply needs to be activated. Add a few seconds and enter it into the Maximum non-critical power fail time. As a good guess, 5-10 seconds might be sufficient. Now keep in mind, during the power failure, the temperature of the chamber and sample will have changed. Depending on the length of the power failure, this temperature change can be small or large. The Freezer has now to decide at what temperature it should continue with the freezing process. For this purpose, we have provided two choices.

• Continue with the freezing process at the temperature of the sample -Sample Temperature. As an example, the temperature of the sample before power fail was at -40°C. After a power fail of 3 minutes, the temperature of the sample increased to -35°C. When power is reestablished, the freezing process will continue with the program run at -35°C up to the last step of the program. This type of setting will provide a smooth temperature change between the actual sample temperature and the freezing process temperature.

• Move to a preset chamber temperature and continue with the freezing process from there. Some applications might require bringing the sample, after a power failure, to a preset temperature. In this case, use the function preset temperature of xx.xx °C.




Seeding (Inducing Ice Formation under Controlled Conditions)

Auto(matic)Seeding (Available as an option) Auto-seeding or Automatic seeding allows the user to select a pre-defined temperature point at which crystallization should begin. Setting a "seed" does this. To be more precise, one point on the outside wall of the sample container (straw, bag or ampoule) is brought to a very deep temperature. The temperature of the medium inside the sample container will also drop rapidly at this point. The temperature of the medium next to this cool spot will have decreased down so far as to induce the crystallization process. The result will be the uniform growing of ice crystals. Most important is the fact that the speed of this uniform crystallization can be controlled. This means that it is possible to control the amount of heat given up within a period of time, giving the system enough time to dissipate the latent heat of crystallization. If the temperature of the sample material or sample solution is far (a few degrees) from the temperature of self-crystallization, performing Autoseeding will result in a slow freezing process. As closer the temperature of the sample is to the point of self-crystallization, as faster the sample will freeze when Autoseeding is turned on, thus being able to control the amount of heat dissipated in time. In addition to Auto(matic)seeding, seeding can be done manually as well. Manual seeding is described in this manual under "Special Functions". Below, you can see a typical autoseeding rack for straws.




Maintenance Leak Check Check occasionally the connections of the Liquid Nitrogen Supply hose for leaks. Due to the high temperature differences in the hose, shrinkage and expansion of the material might cause the hose to become leaky. Just retighten slightly to fix this problem. If the supply system is still leaking, check both sides of the supply hose as well as the connections on freezer and supply vessel. Any damaged parts must be replaced by authorized personnel. Fog (condensed moisture) on the supply tubing and its connections will sink to the ground. This is normal and does not represent a leak in the system. If no liquid is expelled by the supply tube, the system will be tight.

Cleaning of the Freezing Chamber and Instrument Caution: Before doing any cleaning work on the freezer, disconnect it from the power line. Due to technical reasons, the freezing chamber can not be removed from the freezer housing. However, the freezing chamber is fairly large and cleaning should not create any difficulties. Use only mild and soapy non-scratching detergents on a wet cloth. The materials used inside the chamber are Stainless Steel, Glass and Glass fiber reinforced plastics. Use only disinfectants which will not harm those materials. After cleaning and disinfecting, wipe the chamber dry with soft cloth or paper. The freezer housing itself must be cleaned only with a soft wet cloth.

Handling of Temperature Sensors The temperature sensors should be handled with special care. Never apply any force on the sensors. Never try to pull out a sensor from a frozen sample. Wait until the sample reference has changed to liquid again. The sample sensor feed through is marked as blue. The chamber sensor is marked as red. Never try to insert the chamber sensor (red) into a sample. Unknown chamber temperature behavior will be the result. Use only the sample sensor to monitor the sample material. Do not bend or drop a temperature sensor. Limited warranty conditions apply.




Settings on the PC (Done by the instrument administrator)

Disabling of Start-Up Error Message Computers that are delivered from SY-LAB together with an IceCube Series are pre-configured. However, if you supply your own PC or the original PC needs to be changed, some settings have to be made on the PC. In normal operation with the Freezer, no keyboard is connected to the PC. Therefore, the computer has to start without a keyboard. In order to do so, we have to disable in the System-BIOS the entry

"Hold on Keyboard Error " On some computers, this function can not be disabled or is not available in the BIOS. A keyboard error will be displayed on the screen "press F1 to continue". Ignore this message, the computer will still boot up after a few seconds. Note: For BIOS changes, a keyboard has to be connected to the PC.

If this is different on your computer, please refer to the PC`s User Manual.

Disabling Request for Ctrl - Alt – Delete under WINDOWS® 2000 For automatic restart1 of the PC after a power-failure, the login request Ctrl-Alt-Delete has to be disabled.

Press Ctrl-Alt_Delete to begin Ctrl-Alt-Delete helps keep your password secure ----Ctrl-Alt-Del

User name: Administrator

Password: ...................... (leave blank) Log on to: ...................... (leave blank)

□ log on using dial-up connection

OK Cancel Shutdown Option << To remove the above login request, boot up the computer with mouse and keyboard connected. Depending on your Windows® operating system, here we show it for Windows® 2000, go into

Settings - Control Panel - Users and Passwords - Advanced - Secure Boot Setting □ Require users to press Ctrl-Alt-Delete before logging on

Remove the checkmark ( ) from the checkbox and restart you computer again. This time, you will not be asked for Ctrl-Alt Delete anymore.

1 Please Note: Not all computers feature an automatic restart after power loss. For such systems, the PC System must be turned on manually. However, no data will be lost since they are saved internally in the IceCube and are transferred to the PC when power is re-established




However, you will still be asked for a User and Password. Since you did not enter a Password previously, there is no need to enter one. The User name will be already assigned. However, at this point, even though no keyboard is necessary, you still have to press on OK. This would create a problem for the automatic restart feature of the IceCube. Therefore, the Auto Log On feature has to be enabled. Since this procedure will be different among different computers, we suggest leaving this up to your PC system advisor because of changes that have to be done in the registry. Here, we have just given one example.

Caution: Again, we want to point out that these settings should be done only by your PC Advisor. Any

wrong changes made to the registry could lead to total failure of the system. Start - Run - <regedit> (Enter) You are now in the REGISTRY Go to: HKEY_LOCAL_MACHINE \ SOFTWARE \ Microsoft \ WindowsNT \ CurrentVersion \ Winlogon following should be entered: AutoAdminLogon (REG_SZ) = 1 (0 = No automatic Log.on) DefaultDomaineName (REG_SZ) = <Your Domaenname> DefaultUserName (REG:SZ) = <Your Username> DefaultPassword(REG_SZ) = <Your User Password> if one was specified earlier, otherwise, there is no Password entry. Exit

Disabling Power Save Options All power save options (features) must be disabled. These are the Screensaver, no Power down, no Standby and no Hibernation. The disabling procedures vary according from operating system to operator system. Here, we give some examples on how to disable these functions on our currently available XP Professional operating system. Consult the operating system manual for your system.

Windows® XP-Professional • Press the right mouse button while pointing into a free area on the Desktop. • Choose “Properties”. - “Screen Saver” and set the “Screen Saver” field to None. Then, • Click on the button “Power …..” and set all “Settings for ….” to “NEVER”.

e.g. Turn off monitor: Never Turn off hard disk: Never




System standby: Never Sysem hibernates: Never and acknowledge with OK.

Windows® 2000 Prof. • Press the right mouse button while pointing into a free area on the Desktop. • Choose “Properties”. - “Screen Saver” and set the “Screen Saver” field to None. Then, • Click on the button “Power …..”, choose Home/Office Desk as the power scheme and set

all “Settings for ….” to “NEVER”. e.g. Turn off monitor: Never Turn off hard disk: Never System standby: Never While in the “Display Properties” screen, select “Hibernate” and remove the mark from “Enable Hibernate Support”. Now all power save options, etc. should be disabled.




Freezer Software (Re-) Installation (PC knowledge required). Note: Please read all the applicable license agreements. Install only programs for which you are agreeing to these agreements. If you do not agree, do not continue to set up that particular program. Preinstalled programs must only be used when you are fully agreeing to the various program license modules. ............................................................................................................................................

IceCube Series - License Agreement The product IceCube Series (synonyms "program" or "software") is the intellectual property of SY-LAB Geräte GmbH, Austria. It is forbidden to publish, recompile, change or copy the program or parts of it. Diagrams or results obtained with the IIceCube that are the property of the program owner are accepted. For the reason of data safety it is allowed to make one copy of the program CD-ROM. To use the IceCube program, a license from SY-LAB is necessary. It is not mandatory to have an IceCube instrument to obtain a software license. A license is granted as a single place license in form of a hardlock dongle that is necessary to operate the IceCube Series instruments. The loss of this dongle is therefore equivalent to the loss of the license. If only the software CD-ROM gets lost, the license is still valid and a new CD-ROM can be ordered without additional license fees. Free updates are included in the license as long they are aimed to fix possible bugs. Program extensions are not free and are not included in the original license SY-LAB disclaims liability for any loss in connection with direct or indirect usage of the program. The users by themselves are responsible for data safety and data protection COPYRIGHT: ---------- MS Windows 95/98/NT/2000/XP are protected by Microsoft Corp. USA. IceCube (c) 2004 SY-LAB GmbH. Austria. All rights reserved Purkersdorf, 27.5.2004




Every IceCube Series Freezer will be accompanied by a Program CD and a hardware dongle. Note: Before the installation of the software begins, the correct hardware dongle has to be installed either on the parallel printer port of the PC (if a parallel dongle was supplied - older version) or on one of the USB Ports (USB dongle - newer version). Without this dongle, the freezer software can not work properly. Freezers which are delivered with a PC from SY-LAB have already the software preinstalled on the PC. However, the hardware dongle and all cables must still be connected (please consult the section "IceCube Installation - Wiring and Hook-Up" in this manual). Note: To run a validated IceCube Series Freezer program, an IceCubeVal hard lock dongle has to be used. On PC's not supplied by SY-LAB, the software has to be installed as follows: Note: The following freezer software installation assumes that you have a correctly working PC system with Windows® 2000Prof. or XPProf. pre-installed. On systems with Touch Monitor, a keyboard and a mouse is needed during the software installation.

• Make sure the hardware dongle labeled "IceCube Series VAL" is installed. • Insert the Program CD labeled "IceCube Series VAL" into your CD-ROM drive. If the

AutoStart feature is enabled on your PC (please consult the PC-User Manual), the installation of the IceCube program will start automatically. If it does not start automatically, please consult also the file "Readme.txt" provided on the CD for further instructions.

.... if AutoStart for your CD-ROM is not activated or not available, go to INSTALL.BAT located on the IceCube CD_ROM and start it from there. INSTALL.BAT will start/install all necessary

applications and programs.

• First, the hardlock device will be installed. If the hardware dongle is not installed, at program start, you will get an error message

"Error 7: Hardlock not found" • Next, the TouchWare install shield is initialized, then the IceCube install shield program

is loaded. • Before the IceCube Program is installed, you will be asked by the InstallShield to read

the license agreement, choose the destination location and select a program folder. Before the program set-up starts, the current settings like Setup Type, Target Folder and User Information are displayed again. Pressing NEXT will start the installation. Freezer- and ImageLoader Icons will be created after successful installation. Also, you will be notified




that the setup has finished and that you may launch the application by selecting the corresponding icon. Click FINISH to complete the IceCube program setup.

• After the IceCube program installation, you have to go through the TouchWare installation procedure. Press NEXT on the TouchWare welcome screen. Again, you will be asked to read the license agreement. If you accept, press NEXT. The complete Touch Monitor setup is described right in the next section "Touch Monitor and TouchWare Setup".

To start the Freezer program, just press on the freezer icon twice. The Freezer welcome and log-on screen will be displayed. Please consult now the section on "Program Log-In". Please note: In order to restart the IceCube Series Freezer program after a power failure, the freezer -shortcut icon is automatically placed in the PC Desktop - "Startup" menu. PC's that are able to power up automatically should have this feature enabled. Pls consult your PC's operator manual on how to do this or ask you PC system advisor.




Touch Monitor and TouchWare Setup (Only for Variant B to D) Note: In most cases, the touch monitor can be touched with the tip of your fingers. However, some small buttons might be rather difficult to touch. In this case, use the soft backside of a ball point pen, pencil or the pointer of a Palm® (handheld) PC. Do not use any sharp edged devices, otherwise, damage the Touch Monitor Surface might be the result.

Program Installation Computers that are delivered from SY-LAB together with an IceCube are pre-configured. However, if you supply your own PC or the original PC has been changed, the Touch Monitor software has to be newly installed. Also, some settings in the TouchWare software must be set accordingly. Together with the IceCube Series Freezer, you will receive a CD-ROM with all necessary programs and drivers on it. One of these drivers will be the TouchWare driver. Just select the corresponding SETUP Icon. If you have started to install the IceCube software first, you automatically will be guided to the welcome screen below. Following screen will appear.

Go on by pressing Next >. Now you will be asked to accept the License Agreement. Please do so and continue with Next >.




In the next pop-up screen, you will be asked to choose the correct hardware. The Touch Monitors delivered by SY-LAB are connected via the USB port to the PC. Please choose USB and press Next >.

In the next screen, select Express Installation as the default setup and move to the Next > screen.




After the installation, you can take a look at the ReadMe file. Finish will close the Setup program. Note: Please remove the installation disk from your CD-ROM drive before continuing. If you have previously connected the USB cable to the Computer USB port, remove it and plug it back in. The computer system will see that a new device is connected to the USB port. Also, you will be notified that the Touch Monitor has not been calibrated yet. In the following section, we will give you the instruction on how to set up your TouchScreen correctly. Press "Calibrate". You will be presented by a screen showing two dots diagonally arranged. Press and hold shortly each dot (calibration target)at its center. When finished, press "Done". Advance to the following page and follow the instructions.




Setup and Calibration of the Touch Monitor In the above section, we installed the Touch Monitor software. You should now see the TouchWare icon on your desktop area. Open the TouchWare software by selecting this icon. The following screen pops up.

First, select Drawing as the Touch Mode. Next, adjust the yellow Slow and Small Double-click bars as shown on the screen above. Set the Touch Sound Beep to the setting YOU prefer. Here, we set it to Beep on Touchdown. Now move on to the next screen Tools. We will have to move some submenus in Tools before reaching the desired setting. In Tools, select Options. Then move on to Advanced. In the Advanced Touch Screen Settings, select 5 Point (4 Point in some versions) in Calibration Style. Also mark the check boxes: The screens should look like the one shown below.




Pressing twice on Close will get you back to the initial screen.

The Touch Screen Linearize button is now accessible.




Even though the Touch Monitor will work without linearization, we still urge you to perform this task. Linearizing will align more exactly the Touch Monitor to the functions behind it. Press Linearize. Following screen pops up.

There are 25 position spots located on this screen. Try to touch and hold each spot at its center as exactly as possible. After 1 or 2 seconds, each spot will be marked as touched. Follow this procedure for the first 4 rows. On the last horizontal row, try to touch the left 2 spots slightly above. The reason why we should press the spots slightly higher has to do with opening the task bar later on in the Windows® screen. This finishes the setup and installation of the Touch monitor and TouchWare software respectively.




ImageLoader Concurrently to the IceCube program installation, an ImageLoader program was installed. This program will allow you to update the Freezers internal operating program (sub-system). If such an update becomes necessary, SY-LAB or your representative will provide you with a small xxx_yy.IMG file either on diskette or per e-mail. To upload this upgrade file onto the freezer, start the ImageLoader program. Following screen will be displayed.

Select the *.IMG file (in our example - P18_01_V109pA.IMG) and turn the main switch of the 14S Freezer ON. If the freezer was already turned ON, turn it OFF for 2 seconds and ON again. The upgrade file will be uploaded to the freezer. A progress bar, located right under the file name, will advance from the left to the right, indicating the upload progress. The new version of the freezer operating program can then be checked in the Freezer program under Main Menu - Version Info as seen below as an example.




Inserting Your Own LOGO The IceCube software allows you to insert your own company or institute logo. Some space has been reserved for this on the lower right of the screen, just above the SY-LAB Logo. You can only add a logo once. In addition, the SY-LAB Logo is not replaceable The size of the logo should be 106 by 122 pixels in size in bitmap (*.bmp) format. With no prior logo installed, double press on the dark area above the SY-LAB

logo. A file selection screen opens. Enter the name of the logo file you would like to insert and press OPEN. The logo will be placed into the empty black area like shown below. As long as you do not exit the Freezer program, you can replace the logo and play around until the appearance suits you. As soon as you exit the freezer program, the logo file will be permanently displayed.




Troubleshooting Problem Diagnostic / Solution No Power to the system • Check if there is power in the wall socket.

Use a Voltmeter to determine the correct voltage output. Compare the voltage reading with the voltage shown on the freezers type plate.

• Are all components, printer, monitor freezer and PC connected to a power distribution box? Check!

• Is the power distribution box plugged into the wall socket?

• Are all power switches of the single components turned on?

Power to some components but not all

Check the single components for: Power rating (voltage), power connection, broken fuse, power switches turned on?

After turn on, the PC gives error message "Hold on all errors - No keyboard connected"

Enter your PC-BIOS and de-select this function. Check your PC's user manual on how to do this.

Power to PC, Monitor, Printer, and Freezer box ok. IceCube software does not start.

• Message "Error 7: Hardlock not found" shown on screen. The IceCube Freezer Series software is copy protected and needs a "Dongle" to function. Make sure the hardware dongle is installed either on the parallel port (older version) or on one of the USB-ports (newer version). Consult the section on "Freezer Software Installation"

• Windows Screen comes up normally but freezer program still does not start.

• Program fault - re-install the freezer software

• Freezer-Icon on desktop points to a wrong path

Program Log-In not successful 1 • Wrong "User" or "Password". First time users can log-in using for both entries "sylab". The Freezer Administrator should create a new User Name List (also for him) with or without limited access to the system. The factory set Log-In name should then be deleted. See section on "User Administration"

Program Log-In not successful 2

�For first log-in, use sylab for UserName and Password. In addition, the password entry is case sensitive (distinguished between upper and lower case letters). The UserName is not case sensitive.




Password lost / forgotten

�Once individual users are defined, each user has to Log-In using his specific UserName and Password. �In case of a lost / forgotten Password, the System Administrator having Administrator rights can re-assign a UserName and Log-In Password. The Password can then be changed again by the individual user. Thereafter, the Administrator has no way of knowing the users individual password. Note: If the Administrator also forgets his Password, the IceCube Freezer Series Software has to be re-installed. Therefore, it is advisable that two independent persons know the Administrator Password.

Power to PC, Monitor and Printer ok. IceCube software starts ok. "Online (CTS)" LED is OFF.

The freezer electronic (box) is either not turned on, the communication cable to the PC not connected or the freezer box fuse damaged. Check all of the above, replace fuse if needed. Check also the COM port settings in the freezer program - Main - Maintenance - Communication and choose the correct COM Port. Note: When using an USB to RS-232 Adaptor (not part of SY-LAB specification), it will be necessary to set the Adaptor (in the Device Manager) to work on COM1 to COM4 only, besides installing the necessary drivers.

After power failure, the IceCube Program does not start automatically

• Place the shortcut icon in the desktops "Start-up" menu.

• Also, deselect in the BIOS (not everywhere available) the request for a keyboard except on 14S-A, where a keyboard is needed.

• Disable the request for Ctrl-Alt-Del (Win2000). See section on "Settings on PC"

Printer seems not to work After issuing a print command, it might take up to one minute before the printer produces a print-out. Do not touch any printer buttons during this period of time. Other reasons, please check:

• Power connection (power supply) • Printer turned off or not connected • Printer out of paper • Ink cartridge empty • Paper jam

(consult the Printers Operator Manual for further info)

Monitor shows vertical lines after turn on

Sequential turn-on procedure was not followed. Synchronization-problem.




Unplug and reconnect the power plug from the monitor. This will re-synchronize the monitor and should cure the problem.

Touch Monitor not working Assuming the correct software setup of the Touch monitor as described in the user manual, check if the USB connection is properly seated. Unplug shortly the USB cable. Reapply and see if the problem has vanished. Further troubleshooting on PC-System level (see PC Handbook).

No signal to Monitor VGA Monitor cable not connected to either the PC or the Monitor.

Chamber does not get cold during cooling cycle

• No pressure or no LN2 in supply vessel. • Liquid valve on supply vessel turned off. • No supply tube connected between freezer

and supply vessel. • Magnetic valve does not open.

Reasons: Moisture caused MV to get stuck, dirt in system, no electrical signal to valve. Let freezer warm up, try again. If MV still does not operate, an electrical problem might be the cause.

• Supply tube connected to gas-phase of supply vessel instead of to the liquid phase.

Chamber gets cold with no program running

• Magnetic valve slightly leaking - Listen into the chamber compartment. A slight "zzschsch" can be heard. Either the valve seat is dirty or defective. Clean or replace (call service representative)

• Magnetic valve badly leaking - Moisture (Ice) or dirt in the system. Magnetic valve stays open due to one of the above.

• Replace or clean (call service representative) Freezing curve oscillates too much (+/- 3°C)

• Check the supply tube and the pressure vessel for correct pressure. 0.5 to 0.9 bar. If pressure is too low or too high, oscillation can occur.

• Operating conditions not ideally (e.g. supply tube to long or non-standard, supply pressure out of specification, non-standard chamber furniture, etc.

Chamber and / or Sample Sensor Temperature do not follow the Program Curve of the Demo Program

• Various conditions could cause this effect: • no or little pressure in supply vessel (out of

specification) • supply tube connected to the gas-phase of

the vessel • not enough LN2 in the supply vessel • leak in the LN2 supply line • blocked supply line (dirt or ice) • supply tube too long (non-standard)




• temperature sensors defective, dirty or moist • chamber sensor inserted into a sample. The

chamber sensor must never be inserted into anything. It is solely used for regulating the temperature in the freezing chamber

Fan, Magnetic Valve, Heater not working

• Chamber lid not closed, • If physically closed but still get message,

then the electronic does not recognize the lid-switch.

• Contact a SYLAB representative. Heater not working (run any program which has its start temperature at +40°C)

• Heater element broken • Over temperature protection switch open or

defective • Contact a SYLAB representative

Wrong temperature display

• Bad contact on Pt-100 sockets. Check. One of the Pt-100`s might be defect. The resistance of a Pt-100 at about ambient temp. should be around 100 Ohms. Exchange if necessary. The sensors can be switched among each other in order to determine if the fault is on the sensor or the electronic. Thereafter, an Ice-water calibration becomes necessary.

• The chamber is hotter than about 40 to 50°C. The chamber is protected by a temperature switch against over-heating. The heater circuit will be turned off once a temperature of about 40 to 50°C is reached in the chamber. The temperature switch will turn on again once the chamber temperature comes down below 35°C.

Instrument does not cool

• Not enough LN2 in supply vessel. • Not enough pressure in supply vessel (0,5 to

0,9 bar) - (7,5 to 13,5 psi) • Check if the gas phase valve is closed and

that there are no other leaks on the vessel • Supply valve on vessel is closed • Magnetic Valve does not open. Either it is

frozen or there is dirt in the system. In either case, the valve must be removed, cleaned or be exchanged. Should be done only through authorized personnel. Ice particle or dirt might be in the supply vessel. In this case, supply vessel must be exchanged or cleaned.

• Moisture in the system. Never, ever disconnect a cold tube from the system. Moisture from the surrounding air will enter the tube and condense. When warmed up,




water will then be carried to the magnetic valve and will lead to malfunction of the magnetic valve

Instrument does not heat

• Thermo switch open and/or defective • Heater element broken.

Contact SYLAB representative. Log-In not possible Password lost / forgotten

• For first log-in, use sylab for UserName and Password. In addition, the password entry is case sensitive (distinguished between upper and lower case letters). The UserName is not case sensitive.

• Once individual users are defined, each user has to Log-In using his specific UserName and Password.

• In case of a lost / forgotten Password, the System Administrator having Administrator rights can re-assign a UserName and Log-In Password. The Password can then be changed again by the individual user. Thereafter, the Administrator has no way of knowing the users individual password.

• Note: If the Administrator also forgets his Password, the IceCube Freezer Series Software has to be re-installed. Therefore, it is advisable that two independent persons know the Administrator Password.

Pressure display on Monitor (not on 14S) shows different value as the pressure display on the supply vessel (+/- 10%)

• Liquid phase valve on Vessel is closed • During opening and closing of the magnetic

valve (it is normal) • Either the pressure sensor in the freezer or

the one on the vessel are at fault. • Note: The pressure reading does not have to

be very exact. It is more like informing the user that there is pressure applied to the freezer or not.

Temperature Sensor shows erratic values. Spikes on a temperature curve.

• Spikes on a temperature curve indicate a broken temperature sensor. In order to be sure if the temperature sensor or the associated electronic circuit is at fault, exchange the chamber and the sample sensor. If the problem shifts, the temperature sensor is at fault. Exchange the faulty sensor and perform an Ice-Water Calibration.

• Otherwise, there is some electronic problem in the freezer electronic box. Contact a Sylab representative.

"Seeding" does not respond • Seeding option is not installed. Can be up-graded in the field - Ask for quote.




• Seeding valve defective, frozen or dirty - Call Service

• No signal to seeding valve - Call Service • Access to Seeding not granted (User Level)

"Trigger" does not respond (not on 14S Series)

• Trigger not installed • Trigger is malfunctioning - Call Service • Access to Trigger not granted (User Level)

No Alarm output (for all alarms) • Alarm(s) turned off - Check Main Menu - Options - Device Settings. Each alarm can be turned on and off. This applies also to optional external alarms.

• Internal Buzzer defective. Call Service "Start" button does not light up • Chamber temperature is oscillating too much

around the start temperature. See above in this list.

• Device Settings for "Start Temperature reached condition" too narrow or time range to long. Default: +/- 0,5°C for 20 seconds.

LN2 Leak Due to high temperature deviations in the supply tubing and its connections, it will become necessary to check and /or re-tighten both ends of the supply tubing a few minutes after a freezing run has started (when tube is cold).

According to our own findings and the customer's feedback, the troubleshooting list will frequently be updated. Check at: www.sylab.at




Freezer and Accessories Furniture No. Description Description 2 13-158500 Ampoul-rack for 2 u. 5 ml f. 96 pos., made of CrNi 13-158510 Freezing ramp for 36 straws 13-158550 Autoseeding rack f. straws 0,25 ml -43 pcs. 13-158551 Autoseeding rack f. straws 0,5 ml - 35 pcs. 13-158555 Autoseeding rack f. ampouls - 48 pcs. 13-158560 Magazin f. straws 0,25 ml 13-158561 Magazin f. straws 0,5 ml 13-158600 Rack f. Aluminum cassettes 11 positions 13-158515 Quick loading tool 13-158570 Spacer complete 13-158610 Adjustable rack 6 positions 13-158650 Rack f. Cryo Sink SY-Lab reserves the right to change technical specifications without any prior notification.




Spare Parts 14S Pcs. Description Part No. Option Temperature Sensor 65 x 1.0 mm 13-156880 2 Temperature Sensor 65 x 1.5 mm *13-156885 Option Temperature Sensor 100 x 1.5 mm 13-156886 Option Temperature Sensor 160 x 1.5 mm 13-156887 Option Temperature Sensor 200 x 1.5 mm 13-156888 Option Flexible Stainless Steel hose, 1m 13-159010 Option Flexible Stainless Steel hose, 1.3m 13-159013 1 Fan-Motor *13-155400 1 Motor-Axis L43 *13-155411 1 Fan *10-154800 1 Seal for Freezing Compartment S-APF *13-153435 Option Touch Monitor, light grey - RAL 7016 (Variant

B,C,D) *13-156900

1 Magnetic Valve 12V (Note: 1 additional if

Autoseeding is installed) *13-155100

1 Safety Valve 1/4", 50 PSI *13-155080 1 Electronic Box, compl. *13-156000 Option Accessory package - up to 100 Liter Supply Vessel 13-159000 Option Accessory package - over 100 Liter Supply Vessel 13-159100 Option Fuse, 20x5/T 10A, 10 pcs./ pack (for 110V Units) 10-165039-

01 Option Fuse, 20x5/T 5A, 10 pcs./ pack (for 220V Units) 10-165043-

01 Option Additional 3 potential free Alarmrelais 13-159520 *) Included in the standard IceCube14S shipment.




Appendix Statement of Conformity (220 - 240 V AC) Document will be attached to the instrument




Statement of Conformity (110 - 120 V AC) Document will be attached to the instrument




Monitor-Mounting Plate Assembly Depending on the IceCube model that was ordered (Variant B to D) , different monitor accessory packages will be included in the shipment.

In any case, the TFT Monitor is originally packed and needs to be slightly modified in order to fit onto the 14S.

First, mount the swivel heat onto the monitor mounting plate. Tree positions are possible. To position the touch monitor closer to the IceCube main unit, attach the swivel to the leftmost position. To position the monitor further away from the main unit, attach the swivel to the rightmost position. On the left, the center position is shown. Next, screw the mounting plate / Swivel assly. onto

the backside of the TFT Monitor as shown on the following picture.




The vertical angular position of the monitor can be adjusted with the swivel head screw.

Thereafter, plug the swivel head (the end with the red rubber O-ring) onto the monitor base. Monitor base mounting was described earlier in this manual.




Block Diagram - Software

operation manual sylab

Documents