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Pharmacology & Toxicology Core Facilities SOP #:15928.B446.001 Version #:
Author: J. Thompson & T. Szasz Page of 8 Approval Date:
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Core Equipment ID: 15928
Description: BioTek µQuant Microplate Spectrophotometer
Room: B446 (Molecular & Biochemical Core)
Champion:
1.0 Purpose Standardize the process for control, maintenance, ownership and use of the Bio‐Tek µQuant Microplate Spectrophotometer located in room B‐446 Life Sciences. 1.1 BioTek µQuant Microplate Spectrophotometer
Capabilities1 µQuant™ merges the throughput, speed and convenience of microplate formats with the performance of a spectrophotometer. Your laboratory can now perform conventional EIAs at virtually any wavelength, quantitate nucleic acids in the low UV range, or automatically run spectral scans for peak absorbance measurements without using filters. With the patented Bio‐Cell™ accessory, conventional 1 cm measurements can be accomplished. The Bio‐Tek µQuant is an innovative plate reader with a high‐powered Xenon Flash lamp, tunable monochromator and low UV capability.
a. µQuant reads 6‐, 12‐, 24‐, 48‐, 96‐, and 384‐well microplates or 1
cm Bio‐Cells.
b. Pre‐programmed on‐board software addresses a wide variety of applications including nucleic acid quantitation, A260/A280 ratios, pathlength correction, protein analysis, basic ELISAs, and much more.
c. The µQuant truly provides a high‐value solution for unlimited applications.
d. For even more flexibility, control µQuant with Gen5 Data Analysis PC software.
1 µQuant Microplate Spectrophotometer. Bio‐Tek. April 1, 2009. http://www.biotek.com/products/microplate_detection/microquant_microplate_spectrophotometer.html
Pharmacology & Toxicology Core Facilities SOP #:15928.B446.001 Version #:
Author: J. Thompson & T. Szasz Page of 8 Approval Date:
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BioTek µQuant Microplate Spectrophotometer Software Capabilities 2 BioTek’s new Gen5™ software offers a logical interface designed to easily flow from reading parameters, to plate layout, to powerful data reduction, and finally to flexible data output options. The results of this intuitive design, including the exclusive StepWise™ protocol and data reduction tools and the power of Gen5's data reduction, will be evident in increased laboratory efficiency.
2.0 Reason for Issue Maintain a document that describes the Standard Operating Procedures that allows for the standard safe and maximal use of the Bio‐Tek, µQuant Microplate Spectrophotometer within the Pharmacology and Toxicology Core Facilities.
3.0 Process Description Allows Core Facility Users within the Pharmacology and Toxicology Department to properly and effectively use the Bio‐Tek µQuant Microplate Spectrophotometer. The process description details and creates a standard for the use of the Bio‐Tek µQuant Microplate Spectrophotometer. The controlled standard must maintain and adhere to proper and approved research and regulatory qualitative conditions. 3.1 SOP: 15928.B446.001 for LI‐COR Biosciences Odyssey, authored by
Janice Thompson and Theo Szasz, created on April 1, 2009, issued on (insert date).
3.2 SOP: 15928.B446.001 applies to any user and/or trainer of the Bio‐Tek µQuant Microplate Spectrophotometer.
3.3 Responsibilities: All Users are responsible for obtaining the proper approval and training before the use of the Bio‐Tek, µQuant Microplate Spectrophotometer. All users are responsible for the proper use, according to defined protocol, when using the Bio‐Tek, µQuant Microplate Spectrophotometer. a. New Users must ensure their familiarity with the equipment
through thorough reading of this SOP and the associated operating instructions provided by Bio‐Tek. The operating instructions
2 µQuant Microplate Spectrophotometer. Bio‐Tek. April 1, 2009. http://www.biotek.com/products/microplate_detection/microquant_microplate_spectrophotometer.html
Pharmacology & Toxicology Core Facilities SOP #:15928.B446.001 Version #:
Author: J. Thompson & T. Szasz Page of 8 Approval Date:
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manual is located in the closed cabinets on the right side of the Core Facilities Room, B446 Life Sciences.
b. All Users must schedule equipment use in the Equipment Schedule Logbook prior to use. The Equipment Schedule Logbook is located on the desk next to the Bio‐Tek µQuant Microplate Spectrophotometer.
c. All users must record all equipment use in the Equipment Usage
Logbook post‐use on the same day as the recorded use. The Logbook is located on the desk next to the Bio‐Tek µQuant Microplate Spectrophotometer. Within the Logbook on the current log sheet, Users must record the following: Date, PI, Name, Lab Location, Phone Number, Program, Runs, Problems/Comments.
3.4 Equipment Safety Issues3
a. Internal Voltage Always turn off the power switch and unplug the power cord before cleaning the outer surface of the instrument.
b. Potential Biohazards Some assays or specimens may pose a biohazard. Adequate safety precautions should be taken as outlined in the assay’s package insert. Always wear safety glasses and appropriate protective equipment, such as chemically resistant rubber gloves and apron.
c. Liquids Avoid spilling liquids on the instrument; fluid seepage into internal components creates a potential for shock hazard. Wipe up all spills immediately. Do not operate the instrument if internal components have been exposed to fluid.
d. Environmental Conditions Do not expose the system to
temperature extremes. For proper operation, ambient temperatures should remain between 18°‐40°C. Performance may be adversely affected if temperatures fluctuate above or below this range. Storage temperature limits are broader.
e. Sodium Hypochlorite Do not expose any part of the instrument
to the recommended diluted sodium hypochlorite solution (bleach) for more than 20 minutes. Prolonged contact may damage the instrument surfaces. Be certain to rinse and thoroughly wipe all surfaces.
3 µQuant Operator’s Manual, Revision 1. Bio‐Tek.
Pharmacology & Toxicology Core Facilities SOP #:15928.B446.001 Version #:
Author: J. Thompson & T. Szasz Page of 8 Approval Date:
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3.5 Quality Measures
a. Each Use: Each time the Bio‐Tek µQuant Microplate Spectrophotometer is used, clean all exposed surfaces with a cloth dampened with distilled water or distilled water and a mild detergent.
b. Every Six Months: Inspect cables and power cords to ensure their
integrity.
3.6 Example procedure for Lowry protein assay spectrophotometric measurement; BioTek, µQuant Microplate Spectrophotometer Use
a. Turn on the computer from the ON button.
b. Turn on µQuant Spectrophotometer from the ON/OFF switch
on the right side.
c. Double click KC Junior software on the Desktop and click OK on the initialization window.
d. Click on Open Protocol and select the saved protocol from the
drop‐down list. In this case it will be “JLG Lowry”. To create New Protocol for a different assay, refer to the user’s manual, keeping in mind the assay specifics (such as appropriate absorbance wavelength).
e. Click on Modify Protocol. A window will open with the
following tabs: General, Read Method, Template, Curve, Cutoff, Reports. If the saved protocol fits the intended use of the experiment, the only tab requiring modification from default settings will be “Template”. On the “Template” tab, select the appropriate Well Type from the drop‐down list to match the actual well content: “Blank”, “Standard” or “Sample”. To insert selections, click and drag over the 96‐well template area, with sample numbers automatically increasing in a left‐right and up‐down direction. If replicates of the same sample number are included, change the replicate number and indicate their direction (e.g. down and across). To delete selections, select “Clear” from the Well Type drop‐down list and click and drag over the wells that need to be deleted.
Pharmacology & Toxicology Core Facilities SOP #:15928.B446.001 Version #:
Author: J. Thompson & T. Szasz Page of 8 Approval Date:
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f. Click on OK and the software will return to the initial window.
g. Click on Read Plate. A window will open requesting “Results
ID”. Type in the desired name for the reading results.
h. Click on the “Read Plate…” button. A window will open requesting “Plate ID”. Type in the desired name for the plate.
i. Open the lid of the µQuant Spectrophotometer.
j. Remove lid of the 96‐well plate. Insert plate with the A1 well in
the upper left corner.
k. Close the lid of the µQuant Spectrophotometer.
l. Click on “Read Plate”. The spectrophotometer will begin the absorbance measurement of the plate.
m. When scan is done, a window will open displaying the results
(absolute, blanked and calculated values). Check standard curve by clicking on the “Standard Curve…” button, which will open another window. If desired, a copy of the standard curve can be printed from this window. Click OK to close the standard curve window.
n. Under the “Results” menu, select “Print results…” to obtain a
copy of the scan results.
o. To close the results window and exit the KC Junior software, click on the “X” button on the upper right corner.
p. A window will open requesting whether results should be
saved. Make the appropriate selection for saving on a USB flash drive or select “no” if results were printed.
q. A window will open requesting whether protocol should be
saved. Select “No” to keep protocol defaults. For changes to defaults, create New Protocol instead, as needed.
r. Open lid of the µQuant Spectrophotometer. Remove plate and
close the lid of the µQuant Spectrophotometer.
s. Shut down computer and µQuant Spectrophotometer.
Pharmacology & Toxicology Core Facilities SOP #:15928.B446.001 Version #:
Author: J. Thompson & T. Szasz Page of 8 Approval Date:
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t. Sign out in the Equipment Usage Log for the µQuant Spectrophotometer as indicated in Section 3.3c.
3.7 Core Materials
Currently, there are no Core Materials provided for the Bio‐Tek Quant Microplate Spectrophotometer.
3.8 User Materials All materials necessary for the particular equipment applications, including the compatible microplates, are to be provided by users. Users are individually responsible for the compatibility, maintenance and purchase of these materials.
3.9 Use Records
a. Records of Use – All Bio‐Tek µQuant Microplate Spectrophotometer use must be recorded. Refer to 3.3c.
b. Error Messages / System Issues – All error messages and system issues must be relayed to the Equipment Champion and the Pharmacology & Toxicology Core Facility Manager. This information must be relayed on the same day as equipment use. Error messages/system issues must be recorded. Refer to 3.3c.
3.10 Resource Index
Bio‐Tek µQuant Microplate Spectrophotometer literature and resources for the following items may be found at the link below. a. Application Notes (Phosphate and ATPase Determination using
the µQuant Scanning Microplate Spectrophotometer) b. Tech Notes (on Nucleic Acid Quantitation) c. µQuant Single Sheet http://www.biotek.com/products/microplate_detection/microquant_microplate_spectrophotometer.html
4.0 Competences, Authorization and Training
New Users must receive proper authorization from either the Equipment Champion and/or Pharmacology & Toxicology Core Facility Manager before equipment use. A new User may contact the Equipment Champion or Pharmacology & Toxicology Core Facility Manager to schedule training. Training includes SOP and instrument familiarization and any additional required or specialized training. All Users are individually responsible for current SOP familiarization.
Pharmacology & Toxicology Core Facilities SOP #:15928.B446.001 Version #:
Author: J. Thompson & T. Szasz Page of 8 Approval Date:
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5.0 SOP Performance and Equipment Review
The effectiveness of the SOP 15928.B446.001 will be monitored by the Pharmacology & Toxicology Core Facility Manager, Equipment Champion and all Users. Any procedural or qualitative deviations will be reflected within an updated SOP. Any approved User should aptly report any procedural or qualitative issues and/or errors to the Pharmacology & Toxicology Core Facility Manager or Equipment Champion. The Equipment Champion’s name and contact information can be found on the Core Facility Identification Sticker. The Core Facility Identification Sticker is located on the LI‐COR Biosciences Odyssey. The Core Facility Manager’s contact information may be found on the Core Facility Contact Posting within room B446. Updated SOPs will be published and approved Users will be notified. SOP: 15928.B446.001 review will occur every two years. 5.1 Update Date:
Reason for SOP Change: New Version #:
5.2 Update Date Reason for SOP Change New Version #:
6.0 Definitions
Any standard defined term specifically used in the Pharmacology and Toxicology Core Facilities’ SOP. These are terms that carry a unique Core Facility definition that is pertinent to proper SOP understanding and implementation. A list of terms and definitions is maintained to allow Users a quick and efficient reference. Listed below are common Pharmacology and Toxicology Core Facility terms. New common terms are to be approved and added, with definition, to the term reference guide. a. SOP Standard Operating Procedure, which is a standard guide that officially standardizes the process of control, maintenance and ownership of the Bio‐Tek µQuant Microplate Spectrophotometer. The SOP number stand for (xxx . xxx . xxx) equipment number . room number . SOP version number. b. Originator / Author The individual representing the Pharmacology and Toxicology Core Facilities that created SOP: 15928.B446.001. c. Stakeholder Any individual that uses the Bio‐Tek µQuant Microplate Spectrophotometer, including the Pharmacology and Toxicology Core Facilities Department.
Pharmacology & Toxicology Core Facilities SOP #:15928.B446.001 Version #:
Author: J. Thompson & T. Szasz Page of 8 Approval Date:
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d. New User An individual who has not completed the requirements of section 4.0. e. Approved User An individual who uses or performs the task of which is the subject of the SOP and has fulfilled section 4.0. This title may only be given by the Equipment Champion and / or the Pharmacology and Toxicology Core Facilities Manager. f. Champion An individual whose direct expertise with the Bio‐Tek µQuant Microplate Spectrophotometer has been recognized by the Pharmacology and Toxicology Core Facilities Committee. The Pharmacology and Toxicology Core Facility Committee may only award the champion title.
7.0 Appendix The below signatures and dates are required for full SOP approval and implementation. This SOP was written by: Janice Thompson _________________________________ Theo Szasz ________________________________________
This SOP was reviewed by: Dr. Stephanie Watts ______________________________ This SOP was authorized by: Dr. J.R. Haywood __________________________________ Issue Date: _____________________________________