This document discusses blood collection procedures and anticoagulants. It explains that blood can be collected from veins, capillaries, or arteries for various tests like hematological, biochemical, serological, and molecular examinations. The venipuncture procedure and necessary supplies are outlined. Common anticoagulants like EDTA are described which prevent clotting by chelating calcium. Effects of storage temperature and time on blood cell counts and morphology are also summarized.

1. Dr Anupam Singh
SAMPLE
COLLECTION
&
ANTICOAGULANTS
MODERATOR--PROF ASHUTOSH KUMAR

2. •The procedure in
which an operator
bleeds a specific
amount of blood of
subject for a particular
investigation can be
termed as collection of
blood.

3. Common examinations done on
Blood are ???
1.Hematological
2.Biochemical
3.Serological
4.Cultural
5.Molecular

4. Samples can be collected from-
1. Veins- most commonly used.
2. Capillaries
3. Arteries- in case of arterial blood gas
analysis and paediatric patients

5. Venepuncture is a routine procedure .
In order to do this safely the phlebotomist
must have a basic understanding of the
following:
1. Anatomy &
physiology
2.Criteria for
choosing a vein
3.The device to use
4.Skin preparation
5. Personal safety

6. PHLEBOTOMY TRAY---
1. Syringes & needles
2. Tourniquet
3. Specimen containers
4. Request form
5. 70% isopropanol swabs
6. Sterile gauze swabs
7. Adhesive dressings
8. Self sealing plastic bags
9. Rack to hold specimens upright
10. Puncture resistant disposal container

7. Venipuncture
Procedure

10. 1. Prepare the accession order.
2. Approach and identify the patient; sanitize hands.
3. Verify patient diet and latex sensitivity.
4. Assemble supplies.
5. Position patient.
6. Apply tourniquet.
7. Put on gloves.
8. Cleanse venipuncture site.
contd……

11. 9. Perform venipuncture.
10. Order of draw.
11. Release the tourniquet.
12. Place the gauze pad.
13. Remove and dispose of the needle.
14. Bandage the arm.
15. Label blood collection tubes and record time of collection.
16. Send blood collection tubes to proper laboratories.

14. Capillary blood-
• To draw only a small amount of blood
in a microtube
or strip for blood sugar and bleeding
time tests.
• For infants and young children.

17. Why the finger should not be
Squeezed while collecting blood
By finger prick method ?

18. Prick should be deep enough for free flow of
blood or gentle squeezing to start the flow of
blood . Squeezing the finger tip hard results
in tissue fluids diluting the blood , thus
lowering
the hematological values.

19. Why initial 1-2 drops of blood are
discarded in finger prick method ?

20. Initial drops of the blood gets
contaminated with
the antiseptic and therefore
discarded.

21. Why is ring finger preferred
For capillary blood collection ?

22. Ulnar side of the tip of ring
finger is
comparatively less innervated
and
therefore prick is less painful.

23. Reasons to reject specimen for
Hematology are :
A.Clotted specimen
B.Severely hemolyzed specimen
C.Improperly labeled or unlabeled specimen
D.Specimen too old
E.Failure to meet volume criteria
F.Improperly collected capillary specimen
G.Leaking tube
H.Delay in transport
I. Collection of specimen in wrong tube

24. complications
Encountered in
Blood collection

25. Ecchymosis
Syncope
Haematoma
Failure to draw blood
Petechiae
Edema
Obesity
Intravenous therapy
Hemoconcentration
Hemolysis
Burned, damaged, scarred, and occluded veins
Seizures and tremors
Allergies
Inability to obtain a blood specimen

26. What gauze needle should be
used for
collection of venous blood ?

27. •The needles of 19 or 21 G are suitable
for most adults.
• 23G are suitable for children and ideally
should have a short shaft.

28. Order of collection of specimen---
A.Blood culture or sterile tubes (i.e. yellow stopper)
B. Coagulation tube (light blue stopper)
C. Serum tube with or without clot activator or gel
(red, gold or red-gray marbled stopper)
D. Heparin tubes(green or light green stopper)
E. EDTA tubes (lavender stopper)
F. Oxalate/fluoride tubes (gray stopper)

34. ANTICOAGULANT TUBE
EDTA (ETHYLENE DIAMINE TETRA-ACETATE)
•Types: sodium and dipotassium EDTA (1.50-2.2 mg/ml)
•Acts by its chelating effect on the calcium molecules in
blood
•Suitable for routine hematological work such as CBC,
HbA1c
•Repeated inversions of the container the anticoagulant is
thoroughly mixed in the blood added to it.

39. For handling, transfer and storage of
blood samples the following
equipment is needed:
1. transfer and storage tubes (note that some of these should be
freezable)
2. disposable pipettes or pipettes with changeable apex
3. centrifuge. If gel tubes are used, centrifuge should have swinging
bucket rotor
4. timer
5. racks for tubes
6. special boxes for tube transfer and storage
7. set of labels with identification codes or other method to mark the
tubes (note that these should not be vulnerable to freezing)
refrigerator
8. freezer (as required)

40. Effects of storage on blood count-
oVarious changes occur in anticoagulant blood when stored at room
temp and these changes occur more rapidly at higher ambient temp.
oRed cell count, white cell count, platelet count & red cell indices are
usually stable for 8 h after blood collection.
oWhen blood is kept at 4 deg c the effects on blood count are not
usually significant for upto 24 h.
oBest to count TLC & platelets within 24 h and it should be noted
that the fall in TLC may become marked within a few hour, especially
if there is an excessive amount of EDTA.
oRet count are unchanged when blood is kept in either EDTA or ACD
anticoagulant for 24 h at 4 deg c but at room temp count begin to fall
within 6 h.
oHb conc remain unchanged for days.

41. oCoagulation tests be carried out within 2 h when blood or plasma
stored at 22-24 deg c, 4 h at 4 deg c, 2 week at -20 deg c, 6 M at -70 deg c.
oInappropriate handling of blood specimen during transfer to the
laboratory may cause hemolysis, partial coagulation & cell
disintegration.

42. Effects of storage on blood cell morphology-
oOccurs within few hours of colletion. These changes may be discernible
by 3 h & by 12-18 h these become striking.
oSome but not all neutrophils affected, nuclear lobes may become
separated. Small vacuoles appear in cytoplasm.
oIn monocytes, some or many devlop marked changes. Small vacuoles
appear in cytoplasm. Nucleus undergoes irregular lobulation.
oIn lymphocytes, similar changes occur.
oNormal red cells get little affected upto 6 h at room temp but longer
periods lead to progressive crenation & sphering.