HEMATOLOGY 1 LEC

Week 2.1 | BASIC HEMATOLOGICAL METHODS OF EXAMINATION | September 6, 2022

HGB DETERMINATION
● measurement of the concentration of Hgb
BASIC HEMATOLOGICAL METHODS in the blood
OF EXAMINATION ● Protein in RBC that carries oxygen to
HEMATOLOGY TEST your body
● CBC (Complete Blood Count) ● Used also to rule out Anemia
○ a basic screening test and one of ○ Rule out Anemia if low Hgb, Hct
the most frequently ordered ○ If high, polycythemia vera
laboratory procedures.
○ A panel of test that examines the DIFFERENTIAL COUNT
components of the blood ● WBC differential count
■ Hgb, Hct, RBC and WBC ● Percentage of each type of WBC
count, Differential count ○ When B.E.N meets M.ona, L.isa
○ If manual method, ito lang yung mga ● can detect immature WBC, morphology
included (size, shape, inclusion bodies) and
○ If automated, series of tests can be abnormalities, both of which are signs of
made and no need for computation potential issues.
of the indices and checking of ○ Through preparing smears
smear (depends on the protocol) ● RBC and platelets can be seen as well
○ i.e, dinala sa E.R ang patient, may ○ Size and morphology
ubo at lagnat…
HEMATOCRIT / PCV
WHITE BLOOD CELL COUNT ● Volume of packed RBC that occupies a
● Also known as leukocyte count given volume of whole blood.
● We check the actual number of WBCs in a ● PACKED CELL VOLUME (PCV)
liter of blood
● Test that measures the number of WBC in RETICULOCYTE COUNT
your body ● Use to assess the erythropoietic
● Is often included in CBC (production of RBCs) activity of the bone
● A WBC count detects hidden infections marrow
● Helps doctors monitor the effectiveness of ○ Ang ating bone marrow ba ay
chemotherapy or radiation nagre-response pa sa mga kaso ng
○ Pag-nagpapa chemo/ radiation anemia?
therapy, the doctor will request a ○ Capable pa ba mag produce ng
CBC. If the result is all normal, retics si bone marrow?
chemo will push. If the result is low, ○ Meron bang hemolysis kaya ang
no chemo. Or radiation therapy taas ng retics natin?
○ Radiation therapy will affect both ● whole blood, anticoagulated with EDTA is
normal and abnormal cells that is stained with a supravital stain such as
why papataasin muna WBCs prior new methylene blue or brilliant cresyl
to radiation therapy blue , pure azure blue (expensive)
● Treatment in cancer patients ○ The test is 1:1 (1 part specimen, 1
part stain); equal drops dapat
RED BLOOD CELL COUNT ○ Mix then incubate for 50 mins. Then
● blood test used to find out how many / prepare a good smear
how few RBCs you have
● The actual number of red cells per volume of ● SUPRAVITAL STAIN
blood ○ Is used to stain living cells
● also known as Erythrocyte count ○ Reticulocytes still have remnants
● important because RBCs contain Hgb of RNA
(inside) which carries oxygen to your
body tissues ● Retics are still immature cells that can be
○ Increase RBC found in the peripheral blood
■ Polycythemia Vera ○ Dapat ang retics ay within
0.5-1.5%
○ Decrease RBC ○ Babies’ retics. 2-6%
■ Anemia ● After ng reticulocytes, tsaka palang
magiging RBCs

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● These are special tests requested
separately
● Not included in CBC

Erythrocyte Sedimentation Rate (ESR)

● Sedimentation Rate/ sed rate
● Rate at which red blood cells sediment in
a period of one hour.
○ Depends (others get 2 hrs)
○ Wintrobe
■ glass
○ Westergren
■ disposable
● The rate of fall of RBCs. Gaano kabilis mag
settle RBCs in 1hr
○ i.e, patient have a high ESR of 85
mm (if ganito kataas, mabilis mag
settle RBCs)
● High ESR in individuals with rheumatoid
arthritis and tuberculosis
● Common hematology test and
nonspecific measure of inflammation
○ Nonspecific because isasagot lang
sayo ay YES or NO if may
inflammation/ wala
○ It cannot determine the location/
origin of inflammation
○ Another non specific measure of
inflammation is the CRP (C-
Reactive Protein)
■ Hahanapin kung saan ang
inflammation since
nonspecific test

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 HEMATOLOGY 1 LEC
Week 2.2 | QUALITY ASSURANCE PROGRAM | September 6, 2022

QA IN HEMATOLOGY LABORATORY
● intended to ensure the reliability of the
QUALITY ASSURANCE PROGRAM laboratory test.
● QA is the sum of all those activities in which ● The use of control is to determine whether
the laboratory is engaged to ensure that the machine is working properly, the reagent
information generated by the laboratory is working properly, the medical technologist
is correct. is also working properly
● Includes all aspects of laboratory ● The objective is to achieve precision and
activities that affect the results produced accuracy.
from the choice of method, to the
handling of specimens and reporting ● Accuracy is the closeness to the estimated
results. value to the true mean.
○ Extraction palang, preparation pa ● Precision is the reproducibility of a result,
lang dapat perfect na/ maayos whether accurate or inaccurate
○ Try to correlate the results sa
diagnosis/ condition ni patient
● The real purpose is to determine how
correct/incorrect the result is.
● Quality is a measure of excellence or the
state of being free from error
● Lab should be 0% error (ASAP)

● QC (Quality Control)
○ This will minimize errors through
documentation, controls
● QA (Quality Assurance)
○ Ensures that our tests are reliable
with a degree of precision and
assurance

3 MAJOR ACTIVITIES
● PREVENTIVE
○ Pre-Analytical
○ activities done prior to the
examination of the specimen or
sample that is intended to establish
a system conducive to accuracy
testing.
○ i.e, trainings, seminars, phlebotomy
training, running of controls (to
check machines and reagents)

● ASSESSMENT
○ Analytical
○ activities done during testing to
determine whether the test
systems are performing correctly.
○ i.e, calibration

● CORRECTIVE
○ Post-analytical
○ done when error is detected (after
assessment) to correct the system.
■ Recalibration of the
instrument/machine.
■ Kailangan na magbukas ng
bagong reagent..

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 HEMATOLOGY 1 LEC

Week 2.3 | BLOOD COLLECTION | September 6, 2022

BLOOD COLLECTION
SKIN PUNCTURE
● Dermal puncture, Capillary puncture,
Peripheral puncture
● Blood samples collected are called
peripheral blood instead of capillary
blood.
● Sample is a mixture of capillary, venous, ● REASONS FOR CAPILLARY PUNCTURE
and arterial blood with interstitial (tissue) IN INFANTS AND VERY YOUNG
fluid and intracellular fluid. CHILDREN
○ Advantages of skin puncture using
SITES OF SKIN PUNCTURE earlobe
● finger (middle or ring) ■ Less painful
○ Non-dominant hand ■ Less tissue juices
○ Lateral palmar surface contamination
● earlobe ■ Check for abnormalities in
○ (less free nerve endings, less pain WBC and histiocytes
and less tissue juice)
● <1 yr old ○ Disadvantages of skin puncture
○ lateral portion of the plantar ■ Small amount of blood
surface of the heel/toe. ■ Test cannot be repeated
○ Never on the center because the ■ Can’t add more tests
bone might be pierced as well
○ Order of draw for skin puncture

VENIPUNCTURE
● manner of inserting a needle attached to a
syringe to a palpable vein to collect blood
for laboratory testing.
○ Use index finger when palpating
○ Not thumb because it has pulse
● blood sample collected: VENOUS BLOOD
METHODS OF SKIN PUNCTURE ● most widely used blood sample in all
● Heel Stick Procedure (<1 year old) laboratory tests
● Finger Puncture
○ 3 rd or 4th finger of nondominant ● 3 factors involved in a good venipuncture
hand ○ Phlebotomist
● Earlobe Puncture ○ Patient and his/her vein
○ Check for abnormalities in WBC and ○ The equipment needed
histiocytes (less tissue juices/
contaminants) Key notes to remember on Venipuncture
● Angle of needle insertion
● Sites to avoid: ○ Routine venipuncture – 45 degrees
○ inflamed and pallor areas ○ 15-30 degrees
■ No free flowing blood ● Donor bleeding
○ cold and cyanotic areas ● Needle should be bevel up
■ Remedies can be soaking in ● Length of needle
lukewarm water, damp cloth ○ 1 to 1.5 inches
(slight warm) for babies in ● Tourniquet
NICU ○ 1 min (prevent hemoconcentration)
○ congested and edematous areas ● Blood pressure cup
■ No free flowing blood ○ 40 to 60 mmHg (alternative)
○ scarred and heavily calloused areas ● Number of attempts
○ maximum of 2
● Position of the patient
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METHOD OF COLLECTION ● Advantages of ETS
1. Syringe method ○ Faster blood collection
○ Large amount of blood is obtained
○ Blood can be transported and stored
○ Additional and repeated tests can
be done
● Disadvantages of ETS
○ Needs more time and skills
○ Requires more equipment
○ Complication may arise (make sure
patient vein is in good condition)
ORDER OF DRAW-SYRINGE METHOD ○ Not advisable to infants/ small
● Blood culture children and individuals with thicker
● Other sterile tubes skin
● Light blue
● Serum sterile tubes WINGED BLOOD COLLECTION SET
● Green
● Lavender
● Gray
● Yellow
● Red

PARTS OF SYRINGE

● If the vein of patient is fragile

● Check the tubing
● Use micropore as a support/ use your
hands to support the ‘wings’

SITES OF VENIPUNCTURE
● In newborn infants up to 18 months old
○ External jugular vein
○ Temporal vein (scalp vein)
○ Superior longitudinal sinus
1. Tip
2. Barrel (holds the volume)
3. Flange
4. Plunger (pull)
5. Plunger end
6. Needle cover
7. Bevel
8. Gauge number
9. Shaft ● OLDER CHILDREN (18 MONTHS TO 3
10. Needle Hub Y/O)
11. Bevel (slanted part) ○ Femoral vein
12. Tip ○ Long saphenous vein F.L.A.P
13. Lumen ○ Popliteal vein
○ Ankle vein
ETS METHOD (EVACUATED TUBE SYSTEM)

● 3 y/o to adult life

○ wrist vein
○ dorsal vein of the hand
○ dorsal vein of the ankle

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VEINS OF THE ANTECUBITAL FOSSA
● Two patterns of vein
○ “H” pattern
■ Median capital vein (1)
■ Cephalic vein (2)
■ Basilic vein (3)
○ “M” pattern
■ Median (1)
■ Accessory cephalic vein (2)
■ Basilic vein (3)

ORDER OF DRAW FOR VENIPUNCTURE

● Blood culture (yellow stopper)
● Coagulation tube (blue)
● Serum tubes with or without activator
● Heparin (green)
● EDTA (lavender)
● Sodium fluoride tube with or without
EDTA or oxalate (gray stopper)

Complications
● hematoma/ bruise
● Fainting
● Fear of needles
● Allergies
● Hemoconcentration

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 HEMATOLOGY 1 LEC
Week 2.4 | ANTICOAGULANTS USED IN HEMATOLOGY | September 6, 2022

3. Heparin
● Most commonly used for Osmotic Fragility
MOST COMMON ANTICOAGULANTS USED IN Testing and immunotyping
HEMATOLOGY ● Will check the RBCs kung gaano siya
1. EDTA/Versene/Sequestrene tatagal sa different concentrations of sodium
● Most commonly used chloride
● MOA: binds to non-ionized form of ● MOA: binds thrombin
Calcium then chelates Calcium ○ (activated prothrombin)
(important factor for blood coag.) ● 2 forms:
○ Lithium Heparin
● 3 forms: ○ Sodium heparin
○ Dipotassium EDTA
■ Most soluble Sodium heparin
■ Most preferred ● Uses: RBC count, hemoglobin,
○ Disodium EDTA hematocrit, ESR and OFT
○ Tripotassium EDTA
● DISADVANTAGES:
Tripotassium EDTA ○ Not
● Recommended amount: 1.2 mg/dl of blood recommended
● Used for: CBC, ESR, Platelet count and for coagulation
PBS studies
● Can be used for modified Westergren because it
(ESR) affects all
○ 2 ml EDTA blood + 0.5 ml of 3.8% of stages of
0.129 M Na Citrate blood
○ 2 ml of EDTA blood + 0.5 ml NSS coagulation
○ Not for WBC
● DISADVANTAGES: ○ Not recommended for blood
○ Not recommended for coagulation smear preparation
(light blue is used) and platelet ○ Most expensive
satellitism
○ Preparation of blood smear from old 4. OXALATES
EDTA sample changes the ● MOA: binds Calcium
morphology of the red cell (w/in 2 ● Recommended concentration:
hrs) ○ 1-2 mg/ml of blood
● 3 forms of oxalate:
2. CITRATE ○ Double oxalate (commonly used)
● Most common and preferred anticoagulant ○ Lithium Oxalate (bloody body f.)
for coagulation studies ○ Sodium Oxalate (for coag stud.)
● MOA: binds calcium
● Contained in light blue top Evacuated tube Double Oxalate
● Anticoagulant buffer: 3.8% / 0.109 M Na ● salts of Ammonium and Potassium
Citrate ● when used separately:
● PT, APTT (sensitive) ○ Ammonium oxalate (RBC Swells)
○ Potassium oxalate (RBC shrinks)
● Also known as “Balance Oxalate”
● Uses: all RBC
valuation tests, since
there is no effect on
RBC
● DISADVANTAGES:
○ Not
recommended
for blood
transfusions
○ Causes agglutination or clumping of
WBCs and platelets
● Not recommended for peripheral blood
smear preparation
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