Sample Collection

A Laboratory Point of
View
Lt Col AK Harith
Department of Biochemistry
Why do we need to know how to
collect a specimen …

Adequacy of the sample sent

Confounding factors

Concept of Preanalytical variations

 Samples usually sent
Blood Venous
Serum
Plasma
Capillary blood
Arterial Blood
Urine Random sample
24 hrs sample
Timed 24hrs collection
 Misc
CSF

Pleural / Peritoneal fluid

Synovial fluid
 Collection of Blood
 Universal protection
 Plan well before poking needle
 Keep appropriate tubes ready
 Label tubes before hand
 Collect adequate quantity of blood

 AVOID HEMOLYSIS

Causes increase in colour of serum

Values of certain parameters rise
 How to avoid hemolysis
 Use dry containers for blood collection
 Use wide bore needle
 Let the blood flow into the syringe
 While filling the container
remove the needle
avoid frothing
gently mix the anticoagulant if used
 Type of blood samples collected
Serum Most of the Biochemistry tests
Serological tests
Blood grouping and cross matching
Plasma Glucose estimation
Hematological studies
Coagulation studies
Capillary Point of care testing
blood Screening/Testing for Inborn Errors of
Metabolism
Whole ABG studies
blood LE Cell phenomina demonstration
 Serum
Obviously no anticoagulants used

Fasting sample preferred

Chylomicrons cause increase in turbidity of the
serum and affect colorimetric assays

Usually 5ml of blood is collected in a sterile bottle.

Blood is allowed to clot

Whole process takes atleast 2 hrs

before the test can be done
Clot is gently dislodged
Clot retraction takes place
Serum is released
Transferred to an other test tube
Centrifuged
The clear supernatant is used for
further studies
 Plasma
Picture bottle EDTA Hematological
Li/K/Na salts studies
Sugar bottle NaF Plasma glucose
Na iodoacetate
Citrate mixture Sodium citrate Coagulation
studies
Heparin bulb Heparin ABG
OFT
Wintrobes Ammonium and OFT
Mixture Potassium ESR
oxallate
 Mechanism of action
EDTA Chelates calcium ions
Can not be used for Coag studies
Can not be used for biochemistry
NaF Inhibits glycolysis (Enolase)
Iodoacetate Inhibits glycolysis * * * * * * *
Wintrobes Preserves the RBC shape.
mixture Ammonium salts interfere in urea
estimation
Citrate Binds to calcium
Heparin Activates Antithrombin III
Concentration of the Anticoaglulants
Anticoag Conc Vol of blood

NaF 2mg/ml 2 ml

EDTA 2mg/ml 2ml

Citrate 3.2% solution 4:1 for coag

studies
9:1 for ESR
Heparin 10IU/L
 Vacutaniers
 Tubes having anticoag
 Inbuilt vacuum to suck blood
 Draws in adequate volume of blood
 Works on a push concept and hence chances of
counter is low
 Tubes can be directly introduced into the autoanalyser
 Closed system – less chance of blood contamination
 Urine
Random sample : Any time

24 hrs urine sample : AFB culture

Require to prevent bacterial contamination
Preservatives used : HCl, Formalin, Thymol

Timed 24 hrs sample collection :

Clearance tests : Procedure
 CSF
 Precious fluid – cant afford to play with it

 First sample sent for biochemistry

 Second sample for cytology

 Third sample for microbiology

 CSF
No anticoagulants used
Tests done rapidly

Note : Traumatic vs Hemorrhagic tap

Yellow colour of CSF
Protein correction in traumatic tap
CSF sugar takes 6-8 hrs to
equilibriate with serum
 Pleural / Ascitic fluid
No special precautions

Synovial Fluid
Very viscous fluid
Role of anticoagulants felt