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WELCOME

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Presented by:Dr. Priyanka Buragohain

Guided by :Dr Swapan Kr Chakraborty.

Prof. & HODDr Anup BaishyaAssociate Prof.

Dr. Hemen KalitaAssistant Prof.

Dr. Mrinal BaishyaLecturer

SAMPLE COLLECTION AND SAFETYPROCEDURE IN LABORATORY

Why is sample collection necessary ?

To study different investigative parameters

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Blood Urine Stool Sputum Semen Cerebrospinal fluid Body fluids Gastric washings

Ear/Eye/Nose/ Pernasal/Wound/ Throat/ Vaginal swabs

Nasopharyngeal aspirate

Fungal samples of hair, nail & skin etc.

Biopsy material

SAMPLES COLLECTED IN LAB

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Haematological study : Blood R/E, Hb%, CBC count,PBS study, coagulation profile study, genetic study etc.

Biochemical study : Blood sugar, LFT, Bl. Urea, Sr. creatinine, Sr. uric acid etc.

Serology : CRP, VDRL, HIV, RA factor, ASO titre, widal test, etc.

Bacteriological culture Immunological study : detection of antibodies

and antigens, etc. Blood transfusion services : ABO grouping, cross

matching of blood, etc.

STUDY OF DIFFERENT INVESTIGATION FROM A SAMPLE OF BLOOD

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For fasting BS- empty stomach with a preferable fasting of 8-10 hours

For PPBS- a full meal or 75 gm of glucose intake

For lipid profile- a fasting sample of not less than 12 hours, no fatty foods in previous meal

No alcohol before sample collection

PATIENT PREPARATION

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BLOOD COLLECTION PROCEDURE

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Veins : Anticubital vein, veins of arm, dorsum of hands, long saphenous vein, femoral vein, umbilical and scalp vein.

Capillaries :Finger tips, lobule of ear, heel and thumb.

Arteries : femoral artery .

SITES FOR COLLECTION OF BLOOD

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VENOUS BLOOD COLLECTION Make the patient sit Arrange the required equipments Examine the site of collection Locate the site Apply the torniquet Locate the vein Cleanse the area with an alcohol wipe Dry with gauze Feel and fix the vein by pressing down on the vein Remove the needle shield Approach the vein in the same direction the vein

is running with the needle 15* to the participant’s arm

Push the needle with bevel facing up Make sure that the needle is in the vein Loosen the torniquet and allow the needle to fill as

much as needed Withdraw the needle out of the arm, press gauze

firmly on the puncture Collect the blood in approprite container. Discard the needle into a sharps container.

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Wipe the tip of the finger, using alcohol swab and let it dry.

3rd and 4th fingers of non dominant hands are preffered.

Using a sterile lancet, make a skin puncture just of the centre of the finger pad. The puncture should be made perpendicular to the ridges of the fingerprint.

Wipe out the first drop of blood. Free flowing blood may be collected by

gently massaging the finger. Hold a small gauze pad over the puncture

site for a couple of minutes to stop bleeding.

In infant, gently rub the heel to warm it. Clean it and prick on the medial/lateral part of plantar surface to collect blood.

CAPILLARY BLOOD COLLECTION

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For blood gas analysis

ARTERIAL BLOOD COLLECTION

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Red top

ADDITIVE None

MOA blood clots and the serum is separated by centrifugation

USES Chemistries, Immunology and serology, blood bank (cross matching)

Gold top

ADDITIVE None

MOA SST contains a gel at the bottom to separate blood from serum on centrifugation.

USES Chemistries, Immunology and serology

ANTICOAGULANTS

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Light green top

ADDITIVE Lithium heparin

MOA Anticoagulates with heparin. Plasma is separated with PST gel at the bottom of tube

USES Chemistries

Purple top

ADDITIVE EDTA

MOA Forms calcium salts to remove calcium

USES Haematology and blood bank

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Light blue top

ADDITIVE Sodium citrate

MOA Forms calcium salts to remove calcium

USES Coagulation tests

Green top

ADDITIVE Sodium heparin or Lithium heparin

MOA Inactivates thrombin and thromboplastin

USES For lithium level, ammonia level

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Dark blue top

ADDITIVE EDTA

MOA Tube is designed to contain no contaminating metals

USES Trace element testing, toxicology

Light grey top

ADDITIVE Sodium fluoride and potassium oxalate

MOA Antiglycotic agent preserves glucose upto 5 days

USES Glucoses

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Yellow top

ADDITIVE ACD (acid citrate dextrose)

MOA Complement inactivation

USES HLA tissue typing, paternity testing, DNA studies

Yellow black top

ADDITIVE Broth mixture

MOA Preserves viability of microorganisms

USES microbiology- aerobes, anaerobes and fungi

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Black top

ADDITIVE Sodium citrate

MOA Forms calcium salts to remove calcium

USES ESR

Orange top

ADDITIVE Thrombin

MOA Quickly clots blood

USES STAT serum chemistries

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Light brown top

ADDITIVE Sodium heparin

MOA Inactivates thrombin and thromboplastin contains no lead

USES Serum lead determination

Pink top

ADDITIVE Potassium EDTA

MOA Forms calcium salts

USES Immunohaematology

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White top

ADDITIVE Potassium EDTA

MOA Forms calcium salts

USES Molecular/ PCR and bDNA testing

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Adult draw: Min 2 ml 10 ml red top &10 ml lavender top for

antibody 16-20 ml for culturingPaediatric draw : 0.5 – 5ml for culturingNewborn : capillary

SPECIMEN VOLUME

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Wash hands thoroughly with soap and water.

Dry completely Let first stream of urine to drain into toilet. Place a sterile container under the stream

and fill the container. Do not touch the rim or inner surface of the

container. Place and tighten lid on the container. Level and sent to laboratory.

URINE SAMPLE COLLECTION

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Prevent contamination

Send urine within 1 hour for accurate culture result

Can refrigerate for upto 24 hours if delay.

Bagged = BAD, highly unreliable

Voided clean catch= 80% - 90% accurate if perineum well cleaned and caught midstream.

Catheterized = Most accurate and reliable.

Supra pubic aspiration = very very accurate

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Urine specimens for culture should be collected in C&S preservative tubes.

Fill the tube upto the minimum fill line. Mix tube 8-10 times immediately after filling. Morning specimen is preffered. Maintain normal daily fluid intake, avoid

alcohol. In case of 24 hr urine collection do not void on

the collection container. In case of creatinine clearance 1st hydrate the

pt. By administering a minimum of 600 ml of water before the collection.

Avoid coffee, tea and drugs

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Position sample collection paper across the rim of toilet bowl.

Make a bowel movement onto the collection paper.

Avoid mixing with urine or water from toilet. Poke onto stool at six different sites. Collect in a neat, clean, wide mouthed jar. Do not clump, scoop, or fill the tube. Screw it tightly and level it. Store between 2-8 degree or room

temperature

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Transport within 1 hour of collection or refrigerate upto 24 hours.

Warm stools are best for detecting ova and parasites.

Not recommended on patients who have been hospitalisd for >3 days & not admitted with a diagnosis of gastroenteritis.

Specimen should be collected before antibiotic therapy is initiated.

Diarrhoeal stool will always give good results.

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Mouth should be pernished before sample collection.

For fungal culture – 3 consecutively collected early morning specimens are recommended.

For AFB – Collect 3 early morning specimens from a deep cough or 3 consecutively collected specimens, each collectad in 8-24 hour intervals with at least one being an early morning specimen.

Sample should be collected in a sterile disposable, impermeable container.

SPUTUM COLLECTION

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In Adult : L3-L4 level In small children :

L4-L5 level or lower. Morning is preferred

rather than late afternoon or evening.

Always use stylette inside the needle.

CEREBROSPINAL FLUID COLLECTION

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Collected from large joints like knee, ankle, hip, elbow, wrist, and shoulder.

10- 20 ml flud to be obtained in 3 or 4 sterile tubes.

1. Plain tube –gross examination, viscisity, mucin clot test

2. EDTA tube- cell counts and microscopic study.

3. Heparinised tube – microbiologic study4. Fluoride tube - glucose

SYNOVIAL FLUID COLLECTION

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Do not remove more than 1 lt of fluid at a time.

Collect in 3 EDTA tubes.

PLEURAL FLUID COLLECTION

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Collect in 3 tubes EDTA- gross and

microscopic examination

Plain- microbiologic examination

Heparinised- chemical examination.

Should be done under CT scan guidance.

PERICARDIAL FLUID COLLECTION

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Urinary bladder to be emptied Patient to lie in supine or

semi reclined position. Large bore I.V. needle is

introduced in the midway between symphysis pubis and umbillicus.

Needle is connected with a rubber tubing hich drains the fluid into the container.

20-50 ml fluid is sufficient for diagnostic procedures.

PERITONEAL FLUID COLLECTION

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Lie the patient in supine position.

Determine the position of foetus and pocket of amniotic fluid with ultrasound.

Prepare the area (mother’s abdomen)

Anaesthesize the area locally Spinal needle of 20 or 22

gauge is inserted into uterine cavity.

10-20 ml fluid is aspirated.

AMNIOTIC FLUID COLLECTION

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SAFETY PROCEDURES IN LABORATORY

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Remain alert Be cautious while working

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A fire extinguisher should always be handy. Keep sand bucket in the laboratory. Take measures to prevent electrical short

circuiting. No smoking in the working zone of the laboratory. Breakable items should be kept in proper racks

and never at the edge of the working table. Do not suck anything with the mouth, use rubber

teets and bulbs for sucking. Do not place eatables on the working bench. Keep finger nails short

PRECAUTIONARY MEASURES

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At the end of the day clean all working benches with a disinfectant. See that nothing except the required electrical appliance is on.

Dispose all infected material properly. The glasswares should be disinfected with

a suitable disinfectant and be cleaned thoroughly with running water.

Use rubber gloves and a nose mask while working.

Wear a laboratory gown or uniform.

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Examples Storage Safe useFlammable chemicals

Ether, xylene, ethanol, methanol, Romanowsky stain, acid alcohol etc.

• Cool storage• In fireproof

metal box• At ground

level• Less quantity

• Never heat over flame

• Use water bath or electric hot plate

• Flame should be at least 10 feet away

Corrosive chemicals

Strong acids like conc. Sulphuric acid, nitric acid etcStrong alkalis like sodium hydroxide,potassium hydroxide

Store them at low levels

• Never mouth pipette

• Pour at below eye level

• Always add corrosive substance to water slowly

Types of chemicals :

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Examples Storage Safe use

Toxic, harmful & irritating chemicals

Potassium cyanide, iodine, formaldehyde, chloroform, methanol, etc

Store in locked cupboard

• Wear protective gloves

• Wash hands after using them

• Never mouth pipette

Oxidising chemicals

Chlorates, perchlorate, strong peroxide, chromic acid, etc

• Keep away from organic materials and reducing agents

• Keep away from flammable chemicals

Handle with utmost care

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Examples Storage Safe use

Explosive chemicals

Picric acid Store under water

• Do not allow to dry

• Never expose to flame

Carcinogens Benzidine, nitrosamines, nitrosophenols, o-tolidine, o- dianisidine

• Level them as carcinogenic

• Handle with special precautions

• Wear protective gloves, mask, eye shields

• Do not allow contact with skin

• After use wash well with cold water

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ACIDS ALKALIS TOXIC SUBSTANCES HEAT BROKEN GLASS ELECTRIC SHOCK CONTAMINATION BY

INFECTED MATERIAL

ACCIDENTS

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Splashes on the skin : bathe the area with 5% aqueous sodium carbonate.

Splashes in the eye : put 4 drops of 2% aqueous sodium bicarbonate into the eye.

Swallowing of acid : Drink 5% soap solution immediately Gurgle with soap solution Give him 2 whites of egg mixed with

500 ml of milk or water 3 glasses of water Rinse the mouth and lips with 2%

aqueous sodium bicarbonate

ACID BURNWASH IMMEDIATELY WITH WATER

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Splashes on the skin : bathe the area with 5% acetic acid/ vinegar

Splashes in the eye : apply drops of saturated solution of boric acid

Swallowing of alkalis : Make him drink 5% solution of

acetic acid/lemon juice/diluted vinegar

Gurgle with the same 3 glasses of water Rinse the mouth and lips with 5%

acetic acid

ALKALI BURNSWASH IMMEDIATELY WITH WATER

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Send for a physician or qualified nurse

Place the victim in the open air

POISONING

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Severe burns : If splashed with burning flammable

solvent;roll him in a blanket or overall Lay the victim on t5he ground Cover him if he is cold Inform the physician Minor burns : Plunge the area with cold water or

ice water. Apply mercurochrome or acriflavine

ointment Apply dry gauze dressing If infected inform the physician.

HEAT BURN

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Wash the wound and remove the glass pieces.

Apply mercurochrome or acriflavin ointment on the wound

Cover with gauze and adhesive tape.

If it bleeds profusely try to stop bleeding by pressing down on it and refer the patient to a physician.

INJURIES CAUSED BY BROKEN GLASSES

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Wash the wound. If it is not bleeding squeeze hard to make it

bleed Bathe the area with antiseptic lotion. Wash thoroughly with soapy water. Bathe again with antiseptic lotion Refer the patient to the physician. If swallowed wash thoroughly with water

and diluted antiseptic lotion

CONTAMINATION BY INFECTED MATERIALS

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It is rare Put off the main

switch Send for a physician Begin mouth to mouth

respiration immediately

ELECTRIC SHOCK

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