BLOOD COLLECTION perpendicular to the

fingerprint lines.
METHOD - Depth of puncture should
be no deeper than 3.1mm.
SKIN PUNCTURE

Indications:
- For infants less than six (6) months
of age, young children,
chemotherapy patients, the elderly
adults with obesity, burns, or when
IV fluids are flowing into the
accessible veins.

Specimen:
● Capillary blood/ Peripheral ● Decreased RBC count,
blood hematocrit, hemoglobin and
- Mixture of capillary, venous, platelet counts.
and arterial blood with ● Increased WBC count.
interstitial and intracellular - Due to tissue injury caused
fluid. by skin puncture.
- Different from those ● Increase Glucose
obtained on venous - Peripheral blood is not
specimens. preferred in glucose testing
- Flows from tissue (venous blood).
(throughout the body) -
Precautions:
Puncture Site: ● Phlebotomist should carry red,
● Lateral or medial plantar puncture-resistant containers in
(bottom) surface of the heel their collection trays.
- For infants under one (1) ● Do not puncture an area that is
year old. swollen, bruised, infected or
- Depth of puncture should already has been punctured,
not be greater than 2.4mm patients with edema, dehydration
or 2mm. or poor peripheral circulation.
- To avoid damage to the ● Warming can increase the blood
bone. flow sevenfold. The site should be
- To prevent infection like warmed to a temperature no
osteomyelitis. greater than 42 degree C for no
● Palmar surface of the distal longer than 3 to 5 minutes.
portion of the 3rd (middle) or 4th - Use warm-washed cloth/
(ring finger) of the nondominant commercial heel warmer.
hand
- Adult with fragile veins. ORDER OF DRAW FOR SKIN
- Puncture on the finger PUNCTURE
should be made
1 Tube for blood gas
 - Patient and his veins
analysis
- The equipment
2 Slides, unless made from Note: “do not inflict harm to the patient”
specimen in the esp. To leukemia patients.
EDTA microcollection
tube Equipment:
3 EDTA microcrollection ● TOURNIQUET
tube - Used to provide a barrier
against venous blood flow
4 Other microcollection to help locate a vein.
tubes with anticoagulants - Should be applied 3 to 4
5 Serum microcollection inches above the
tubes venipuncture site and left
on for no longer than 1
minute.
Note: EDTA comes first than the other
- When bp cuff is used as a
microcollection tubes with
tourniquet, it should be
anticoagulants because adequate
inflated at approx:
volume for hematology should be
______________________
needed in CBC.

● NEEDLES
- The gauge number of a
ORDER OF DRAW FOR SKIN needle is inversely related
PUNCTURE to the bore size.
LAVENDER - Needles used in evacuated
1 EDTA
tube systems are double
2 GREEN Lithium heparin pointed.
❖ LONGER END:
Other LIGHT Lithium heparin ❖ SHORTER END:
plasma GREEN with gel
- Purpose of rubber sleeve in
additives GRAY Sodium fluoride
and potassium the shorter end of the
oxalate needle: _______________
- Standard needle for blood
3 RED No additive collection: _____________
- Most widely used needle
Serum GOLD With clot
tubes activator and gel lengths: _______________
- The needle should be in
bevel position, and should
VENIPUNCTURE
make an approximate 15
degree angle with the arm.
Introduction:
● Easiest and most convenient
method of obtaining enough
volume of venous blood suitable
for a variety of tests.
● Three (3) factors involved in good
venipuncture:
- The phlebotomist
 evacuated tubes, it
● NEEDLE HOLDERS is necessary to
- Needles and holders have multiply the
safety features to prevent obtained platelet
accidental needle stick. count by 1.1 for
accuracy
● COLLECTION TUBES - Excessive concentration of EDTA
- Can be plastic or glass may cause:
- OSHA recommends the ❖ Cell shrinkage leading to
use of plastic tubes decreased spun hematocrit,
whenever possible increased MCHC and a
- Most of the glass tubes are falsely low ESR
coated with silicone ❖ Degenerative changes in
WBCs and platelets will
❖ Additives in Collection Tubes swell and break causing a
- Clot activators falsely increased platelet
- Anticoagulants count
- Antiglycolytic agent ❖ Hgb will NOT BE
AFFECTED
● CLOT ACTIVATORS - EDTA cannot be used for
- Accelerate the clotting coagulation studies
process and decreases the ❖ Interference with factor V
specimen preparation time and the thrombinfibrinogen
- Glass or silica particles reaction

● ANTICOAGULANTS SODIUM CITRATE

Ethylenediaminetetraacetic Acid ● Light Blue
(EDTA) - 1 part of .109M (3.2%) Na
- Normal concentration: _________ citrate to 9 parts of blood
- Lavender (plastic):K2 EDTA/Spray - For coagulation studies
dried - When the Hct of patient is
- Lavender (glass) :K3 EDTA in >/= 55%, the amount of N
liquid form citrate should be decreased
- Pink: Spray dried K2 EDTA ● Black
- Action: Prevents coagulation by - 1 part of citrate to 4 parts of
chelating calcium in the blood blood
- Platelet satellitosis/Platelet - For original/standard
satellitism Westergren ESR method
❖ Adherence of platelets - Action: Prevents
around neutrophils, coagulation by binding the
producing a ring or calcium of the blood in a
satellite effect soluble complex
❖ Due to serum factor which
reacts in the presence of HEPARIN
EDTA - Chemistry tests, blood gas
❖ Remedy: _____________ analysis, EOFT, cardiopulmonary
➢ Because of the bypass
dilution in the citrate - Concentration: 15units/mL of blood
- Used as an in vitro and in vivo
ORDER OF DRAW FOR
anticoagulant VENIPUNCTURE
- Action: Prevents clotting by binding
to antithrombin in the plasma and 1 Blood culture tubes
inhibiting thrombin and activated
2 Coagulation sodium citrate
coagulation factor X
tube
- Caution:
❖ When blood smear is 3 Serum tubes with or without
prepared from heparinized clot activator or gel
specimen and Wright separator
stained, a blue-colored
4 Heparin with or without gel
background may be
obtained 5 EDTA tubes
❖ Never used for
coagulation studies 6 Glycolytic inhibitor tubes
because of its inhibitory
effect on thrombin Venipuncture Site:
● Preferred vein:
OXALATE ● Second choice:
- Concentration: 1-2mg/mL of blood ● Third choice:
- Commonly added to tubes ● Alternative site: If veins cannot be
containing glucose preservatives to found in either arm, it might be
provide plasma for glucose testing essential to assess the veins on
- Action: Acts by formation of the dorsal side of the wrist and
insoluble complexes with calcium hand.
● The veins in the feet should not be
SODIUM FLUORIDE used without permission of the
- Preserves glucose for 24 hours physician.
- Glucose, lactic acid, blood alcohol
- Combined with potassium oxalate
if anticoagulation is needed
- Action: Inhibits glycolysis

SPECIAL-USE ANTICOAGULANTS
● Sodium Polyanetholesulfonate
(SPS)
- for blood culture collection
● Acid Citrate Dextrose
- for immunohematology
tests (e.g. DNA testing,
HLA phenotyping for
paternity testing and
transplant compatibility)

Solution for Skin Antisepsis:

● 70% isopropyl alcohol in a
commercially prepared pad
● The current CLSI standard states
that cleansing with a
back-and-forth scrub is acceptable.

HEMATOLOGY
LABORATORY APPARATUS
TOURNIQUET
- to make the veins more noticeable
and accessible by increasing
venous fullness.
- The tourniquet should never be
worn for more than one minute.
- This could lead to
hemoconcentration or a change in
the results of a blood test.
SYRINGE
- to take blood from patients whose
veins are delicate or tiny.
- A butterfly needle is an additional
device that can be utilized in these
circumstances.
ETHYLENEDIAMINE TETRA ACETIC
ACID (EDTA)
- EDTA is used in concentrations of
1.5 mg/1 mL of whole blood.
- The mode of action of this
anticoagulant is that it removes
ionized calcium (Ca2+) through an
irreversible process referred to as
chelation. This process forms an
insoluble calcium salt that prevents
blood coagulation
- EDTA is available into three forms:
Dry form (Na2 EDTA, and K2
EDTA) & Liquid form (K3 EDTA)
- It is the anticoagulant of choice in
HEMATOLOGY
- Tri-potassium (K3 EDTA) salt also
known as SEQUESTRENE. It is
the liquid form.
- K2 EDTA are recommended by the
International Council for
Standardization in Hematology
(ICSH) and CLSI as the
anticoagulant of choice for blood
cell counting and sizing because
they produce less shrinkage of
RBCs and less of an increase in
cell volume on standing. -Turgeon
- Liquid tripotassium EDTA is often
preferred to the powdered form
because it mixes more easily with
blood- Rodak’s
- For routine cell count and blood
smear preparation
- It is the preferred anticoagulant for
platelet count and evaluation of
blood cell morphology
TUBE RACK
- used to hold upright multiple test
tubes at the same time.
PLASTIC DROPPER
- They are often used to avoid
cross-contamination.
GLASS SLIDE
- to hold objects for examination
under a microscope.
 CELL COUNTER

- used for calculating the

concentration of cells for adjusting
molecular biology experiment
reagents, examining the growth
rate of microorganisms, and
calculating the fraction of dead to
live cells to measure cell viability.

THOMA PIPETTE TALLY COUNTER

- used to prepare leukocyte and - counting liquid droplets, bacterial

erythrocyte microscopic counts. colonies, blood cells, or any other
repeated occurrence.

MICROSCOPE

- utilized to investigate, identify,

treat, and prevent illnesses
affecting the blood and its
constituent parts, such as blood
cells, hemoglobin, blood proteins,
and blood coagulation.
- A hematology microscope is used
to examine the bone marrow and
blood.
- to study blood, blood problems,
HEMOCYTOMETER and blood diseases.
- Blood clots, hemophilia, leukemia,
sickle cell anemia, lymphoma, and
- a device used for manual cell blood parasites are a few of these
counting that is also known as a illnesses and conditions.
haemocytometer or a cell counting
chamber. CAPILLARY TUBE /
MICROHEMATOCRIT TUBE

NEUBAUER CHAMBER
- counting liquid droplets, bacterial
colonies, blood cells, or any other
repeated occurrence.

MICROSCOPE
 - sophisticated instruments designed
to streamline the analysis of blood
- are employed to calculate the
samples.
proportion of red blood cells in
blood by volume. - These machines utilize advanced
technology to accurately count and
characterize the different
SEALER
components of blood, providing
comprehensive data for diagnosis
- To avoid any leaks, the capillary and monitoring
tube's sealing must be strong.
- To prevent inaccurate findings, the
centrifuge machine shouldn't be ERYTHROCYTE SEDIMENTATION RATE
opened during the test. (ESR)
- If the centrifugation is halted, the
likelihood of a mistake in the
outcome will rise. - It is a blood test.
- It measures how quickly
MICROHEMATOCRIT CENTRIFUGE erythrocytes, or red blood cells,
separate from a blood sample that
has been treated so the blood will
- used to calculate the blood's
not clot.
erythrocyte percentage by volume.

MICROHEMATOCRIT READER

- to separate tiny amounts of blood

and solutions and to calculate the
volume fractions of erythrocytes
(red blood cells) in blood.

AUTOMATED MACHINE