This document provides an overview of laboratory procedures for collecting and handling various clinical specimens. It discusses the appropriate collection containers, requirements, and procedures for collecting blood, urine, stool, sputum, cerebrospinal fluid, and other specimens. Specific topics covered include blood collection by venipuncture and fingerstick, urine collection and testing parameters, stool collection for culture and ova/parasite examination, and safety practices for handling hazardous materials in the laboratory.

1. WELCOME
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2. Presented by:
Dr. Priyanka Buragohain
Guided by :
Dr Swapan Kr Chakraborty.
Prof. & HOD
Dr Anup Baishya
Associate Prof.
Dr. Hemen Kalita
Assistant Prof.
Dr. Mrinal Baishya
Lecturer
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3.  To study different investigative parameters
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4.  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
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5.  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.
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6.  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
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8.  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 .
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9.  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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10.  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.
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11.  For blood gas analysis
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12. 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
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13. 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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14. 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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15. 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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16. 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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17. 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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18. 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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19. White top
ADDITIVE Potassium EDTA
MOA Forms calcium salts
USES Molecular/ PCR and bDNA testing
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21. Adult draw:
 Min 2 ml
 10 ml red top &10 ml lavender top for
antibody
 16-20 ml for culturing
Paediatric draw :
 0.5 – 5ml for culturing
Newborn :
 capillary
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22.  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.
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23.  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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24.  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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25.  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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27.  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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28.  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.
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30. 30

31.  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.
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32.  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 study
4. Fluoride tube - glucose
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33. 33

34.  Do not remove more
than 1 lt of fluid at a
time.
 Collect in 3 EDTA
tubes.
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35.  Collect in 3 tubes
 EDTA- gross and
microscopic
examination
 Plain- microbiologic
examination
 Heparinised- chemical
examination.
 Should be done under
CT scan guidance.
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36.  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.
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37.  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.
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38. 38

39.  Remain alert
 Be cautious while working
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40.  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
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41.  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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42. 42

43. Examples Storage Safe use
Flammable
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
etc
Strong alkalis
like sodium
hydroxide,potas
sium hydroxide
Store them at
low levels
• Never mouth
pipette
• Pour at below
eye level
• Always add
corrosive
substance to
water slowly
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44. 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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45. 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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46.  ACIDS
 ALKALIS
 TOXIC SUBSTANCES
 HEAT
 BROKEN GLASS
 ELECTRIC SHOCK
 CONTAMINATION BY
INFECTED MATERIAL
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47.  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
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48.  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
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49.  Send for a physician or qualified nurse
 Place the victim in the open air
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50.  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.
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51.  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.
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52.  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
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53.  It is rare
 Put off the main switch
 Send for a physician
 Begin mouth to mouth
respiration
immediately
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