College of Pharmacy &

Medical Science
Medical Laboratory Sciences
Clinical Hematology &
Coagulation Lab

Instructor: Mahmoud Ramlawi, MSc

LAB #1
Phlebotomy (blood collection)
Phlebotomy
● Is surgical opening or puncture of a vein to withdraw blood or to introduce fluid.
There are two types of blood samples: venous blood and capillary blood.

● Collection of blood sample by Venipuncture : It is one of the most critical phases

of obtaining accurate laboratory test result

● Objectives:
1. To Know the different types of needles, syringes and blood collecting tubes and
their use/
2. To learn how to collect blood samples by different methods.
phlebotomy anatomy
 Equipment
1. Syringes :
• In the past syringes were the only practical way blood could
be collected . They were made of :
• A. Glass : and after each use ,they were placed in paper
bags or cloth wraps and sterilized(by boiling) for reuse.
These have been replaced By:
• B. Plastic , disposable syringes
• C. Evacuated System :tubes and needle holders
Syringes are still used for collecting types of samples:
A. Arterial blood Gases
B. Blood cultures
C. various body fluids
• These are best for patients with small veins , veins that easily
collapse ( when exposed to the pressure in the evacuated tubes )
• They may also be the best choice when other collecting devices
are used ,such as Butterfly infusion sets and intravenous infusion
sets .
Two advantages for syringes use :
1. Blood usually appear in the hub of the syringe as a vein is
penetrated , which is helpful for the phlebotomist and when
collecting blood from arms with fragile , deep or mobile veins .
2.Blood –flow rate and withdrawal pressure is easily controlled by the
phlebotomist , so that collapse of thin walled veins can be
avoided . Large amounts of blood (up to 60 mL ) can be
withdrawn using a small diameter needle and syringe.
2. Needles :
2. Needles :
The Gauge (diameter ) and Length of the needle used on a syringe or
evacuated system depends on :
1.Amount of blood to be drawn .
2.Type of analysis to be performed
3.Condition of potential phlebotomy sites

Its gauge number gives the bore size , or diameter of a needle . The
smaller the number , the greater the diameter, For ex: G24&G26
indicates small needle Lumen(diameter) while , G16&G18 indicates
needles with large lumen.
2. Needles :
● When an evacuated system is used , tubes for collection should be
in the order : 1. Sterile blood culture.2.Tubes without additives
(serum ) .3.Na citrate or Heparinized tubes. 4.Tubes with other
additives (EDTA, Flouride)
3. Tubes :
3. Tubes :
• EDTA (ethylene diaminetetraacetic acid ) (sequestrene, Versene):
• It is used as a disodium or dipotassium salt that prevent coagulation by chelating or
binding Calcium in the plasma . Calcium is required in many steps of the coagulation
process. About 1-2 mg of EDTA is needed for each mL of blood .
• EDTA is the most common anticoagulant used in routine hematological studies .

Advantages :
a. Prevents Platelet clumping .
b. Preserves cellular morphology.
c. Prevents artifacts in preparing blood films .
d. Little effect on chemistry tests
e. EDTA blood stored at 4c for 24hrs . If tested again show no changes in results compared to the
previous results .
Disadvantages :
a. Inhibits Alkaline phosphatase
b. Not useful for Calcium and Iron testing
c. If in excess : shrink RBCs ,so dec . Hematocrit , False ESR results and MCHC . , Degenerates
WBCs & platelets so inc. platelet count.
• Heparin : inactivates Fx and FII(thrombin).

• Available as Sodium , Potassium and Ammonium salts , It inhibits Thrombin action

.About 20 units of heparin are required to anticoagulated 1 mL of blood .
• Advantages :
1. minimum interference in most chemical tests .
2. Available as a liquid or a powder .
3. can be used in Phosphorous test.
• Disadvantages :
1. Relatively expensive .
2. Produces blue background on Wright –Stained smear .
3.Inhibits acid Phosphatase activity .
4.Causes clumping of WBCs & Platelets .
Sodium Citrate : this is the anticoagulant of choice for the coagulation studies . A concentration
of 3.2% is used , mixing 1 part of Na citrate with 9 parts of blood . Calcium is chelated ,
which is easily reversed by the addition of ionized Ca since it is required to clot the plasma
in coagulation tests .
Advantages :
a. Maintain stability of coagulation factors (V&VIII&platelets)
Disadvantages :
a. Not suitable for many chemistry tests .
b. b. Inhibits Alkaline Phosphatase .
c. c. Cause distortion of cells morphology .
2- Capillary Blood Capillary blood is used for infants under 1 year
of age and when it is not possible to obtain venous blood.
Common sites for drawing capillary blood are the heel and the ball
of the middle finger.
Areas to Avoid When Choosing a Site for Blood Draw:
Certain areas are to be avoided when choosing a site for blood draw:

1- Extensive scars from burns and surgery - it is difficult to puncture the scar tissue and obtain a
specimen.

2- Hematoma - may cause erroneous test results. If another site is not available, collect the
specimen distal to the hematoma.

3- Intravenous therapy (IV) / blood transfusions - fluid may dilute the specimen, so collect from
the opposite arm if possible.

4-Cannula/fistula/heparin lock - hospitals have special policies regarding these devices. In

general, blood should not be drawn from an arm with a fistula or cannula without consulting the
attending physician.

5- Edematous extremities - tissue fluid accumulation alters test results.

Techniques to Prevent Hemolysis (which can interfere with many tests):
1- Mix all tubes with anticoagulant additives gently (vigorous shaking can cause
hemolysis) 5-10 times.
2- Avoid drawing blood from a hematoma; select another draw site.
3- Make sure the venipuncture site is dry before proceeding with draw.
4-Avoid milking the site when collecting capillary samples
5- Do not use a small diameter needle if not needed.
6-Avoid a probing, traumatic venipuncture.
7-Avoid prolonged tourniquet application (no more than 2 minutes; less than 1 minute is
optimal).
8-Avoid massaging, squeezing, or probing a site.
9-Avoid excessive fist clenching.
10- If blood flow into tube slows, adjust needle position to remain in the center of the
lumen
 Evacuated System :
• The most common blood collecting system use: 1.A disposable
needle . 2.A needle holder .3. An evacuated tube

• The needle screw into the holder and each end of the needle is
pointed ; one to enter the vein ; the other is to puncture a collecting
tube . The vein is entered first , and then any number of collecting
tubes can be added to the other end . Each tube is punctured through
the rubber cap and blood flows into it due to the partial vacuum .
(gradually loss of vacuum from the tubes that have been stored on
shelf a few years .
Advantages of the Evacuated System :
1. One needle holder can replace many bulky syringes saving space on Phlebotomy
trays .
2. A number of collecting tubes with various additives can be used sequentially (some
patients experience anxiety at the sight of a 30 mL syringe being filled with their
blood whereas 5-6 tubes may not seem as threatening )
3. Blood enters each tube and mixes rapidly with a specific anticoagulant , whereas
blood in a syringe must either forgo anticoagulant until the phlebotomy is complete ,
or the syringe must be coated with anticoagulant , thus limiting the variety of
additives used.
4. Blood collection is faster with the evacuated tubes than with the syringes especially if
several tubes are to be filled .
Blood parts