HEMATOLOGY – LABORATORY

PRELIM TOPIC 1. COMPLETE BLOOD COUNT

Lecturer/s: Ms. Lee-an Anayon, RMT, Mrs. Lorraine Mission-Maravilla, RMT, Mrs. Regine Morados, RMT
Source/s: Rodak’s Hematology: Clinical Principles and Applications

INTRODUCTION: BLOOD
➔ The average human possesses 5 liters of blood
Out-patient Department In-Patient Department
FUNCTION OF BLOOD: ● Done in the extraction ● Done in bedside of
● Transports oxygen from lungs to tissues area patient
● Clears tissues of carbon dioxide ● A dedicated ● ER, OR, Wards, ICU
● Transports glucose, proteins, and lipids phlebotomy chair is ● Some hospitals
● Moves wastes to the liver and kidneys. present. establish warding
➔ The liquid portion is plasma ● Also known as time for the
“walk-ins” phlebotomist to
● Plasma provides coagulation enzymes that
● No collected blood
protect vessels from trauma and maintain schedule/appointment ○ Not unless STAT
circulation. is needed sample is
● transports and nourishes blood cells needed.

THREE CATEGORIES OF BLOOD CELLS ➔ Accessioning

1. Red blood cells (RBCs) or erythrocytes ● The specimen must be accurately
2. White blood cells (WBCs) or leukocytes registered in the work list:
3. Platelets (PLTs), or thrombocytes ❖ Accession may be automated,
relying on bar code or
➔ Hematology is the study of these blood cells radiofrequency identification
● By expertly staining, counting, analyzing, technology, thus reducing
and recording the appearance, phenotype, instances of identification error.
and genotype of all three types of cells, the ❖ Once it is transmitted to the
medical laboratory professional is able to machine, you can see the results.
predict, detect, and diagnose blood ➔ Processing
diseases and many systemic diseases that ● most laboratories employ automated blood
affect blood cells. cell analyzers to generate the CBC.
● Physicians rely on hematology laboratory ● Many blood cell analyzers also provide
test results to select and monitor therapy comments on RBC, WBC, and platelet
for these disorders; consequently, a morphology
complete blood count (CBC) is ordered on ● When one of the results from the blood cell
nearly everyone who visits a physician or analyzer is abnormal, the instrument
is admitted to a hospital. provides an indication of this, sometimes
COMPLETE BLOOD COUNT (CBC) called a flag.
➔ is performed on automated blood cell analyzers and ❖ In this case, a “reflex” blood film
includes the RBC, WBC, and platelet examination is performed
measurements ❖ A medical technologist should
● commonly used in routine blood checkup know how to manually process
● baseline test each parameter in order to
LABORATORY WORKFLOW IN PERFORMING counter-check flags
COMPLETE BLOOD COUNT ● These measurements include:
➔ Specimen collection
● The medical laboratory professional
collects a blood specimen for CBC. He/she
must ensure that:
❖ The specimen must be of
sufficient volume, because “short
draws” result in incorrect
anticoagulant-to-blood ratios
❖ The specimen must be tested or
prepared for storage within the
appropriate time frame to ensure
accurate analysis
Out-patient department/ In-patient department
➔ Releasing of results

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 ◆ Should there be abnormalities in the
result, or such circumstances where
results are too high or too low compared
to the reference ranges, it is just that the
medical technologist should perform
Manual CBC.
BLOOD COMPONENTS
(A), Erythrocyte (red blood cell, RBC); (B), neutrophil; (C),
band (D), eosinophil; (E), basophil ; (F), lymphocyte (G),
monocyte; (H), platelet.
RED BLOOD CELLS
➔ RBCs are anucleate, biconcave, discoid cells filled
with a reddish protein, hemoglobin, which transports
oxygen and carbon dioxide
➔ appear salmon pink and measure 7 to 8 mm in
diameter with a zone of pallor that occupies
one-third of their center, reflecting their biconcavity
➔ Since before 1900, physicians and medical
laboratory professionals counted RBCs in
measured volumes to detect anemia or
polycythemia.
● Anemia means loss of oxygen-carrying
capacity and is often reflected in a reduced
RBC count or decreased RBC hemoglobin
concentration
● Polycythemia means an increased RBC
count reflecting increased circulating RBC
mass, a condition that leads to
hyperviscosity
➔ Historically, microscopists counted RBCs by
carefully pipetting a tiny aliquot of whole blood and
mixing it with 0.85% (normal) saline
● Normal saline matches the osmolality of
blood; consequently, the suspended RBCs
retained their intrinsic morphology, neither
swelling nor shrinking
➔ A 1:200 dilution was typical for RBC counts and a
glass pipette designed to provide this dilution, the
Thoma pipette, was used routinely until the advent
of automation.
➔ The diluted blood was transferred to a glass
counting chamber called a hemacytometer
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HEMOGLOBIN DETERMINATION
➔ Hemoglobin measurement relies on a weak solution
of potassium cyanide and potassium ferricyanide,
called Drabkin reagent
➔ An aliquot of whole blood is mixed with a measured
volume of Drabkin reagent, hemoglobin is
converted to stable cyanmethemoglobin
(hemiglobincyanide)
● the absorbance or color intensity of the
solution is measured in a
spectrophotometer at 540 nm wavelength
➔ The color intensity is compared with that of a known
standard and is mathematically converted to
hemoglobin concentration.
HEMATOCRIT DETERMINATION
➔ also called packed cell volume (PCV)
● packed cells referring to RBCs
➔ the ratio of the volume of packed RBCs to the
volume of whole blood
➔ manually determined by transferring blood to a
plastic tube with a uniform bore, centrifuging,
measuring the column of RBCs, and dividing by the
total length of the column of RBCs plus plasma.
➔ The normal ratio approaches 50%
➔ The buffy coat is a light-colored layer between
RBCs and plasma and contains WBCs and platelets
● excluded from the hematocrit
determination.
RBC INDICES
➔ The medical laboratory professional may use the
three numerical results
● RBC count, HGB, and HCT, to compute
the RBC indices mean cell volume (MCV),
mean cell hemoglobin (MCH), and mean
cell hemoglobin concentration (MCHC)
➔ Extreme RBC volume variability is visible on the
Wright-stained blood film as variation in diameter
and is called anisocytosis
➔ RBC indices provide stable measurements for
internal quality control of automated blood cell
analyzers
➔ All these parameters are employed to detect,
diagnose, assess the severity of, and monitor the
treatment of anemia, polycythemia, and the
numerous systemic conditions that affect RBCs
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 ➔ Automated blood cell analyzers are used in ➔ In the Wright-stained blood film, 0.5% to 2.5% of
nearly all clinical laboratories to generate these RBCs exceed the 7- to 8-mm average diameter and
data, although visual examination of Wright-stained stain slightly blue-gray.
blood film is essential to verify abnormal results. ➔ polychromatic (polychromatophilic) erythrocytes are
newly released from the RBC production site: the
INDEX DESCRIPTION FORMULA
bone marrow
MCV Mean corpuscular volume HCT/RBC x ➔ are closely observed because they indicate the
● reflects RBC diameter 10 ability of the bone marrow to increase RBC
on a Wright-stained production in anemia caused by blood loss or
blood film excessive RBC destruction
(femtoloters/ fL) ➔ Methylene blue dyes, called nucleic acid stains or
vital stains, are used to differentiate and count
MCH Mean corpuscular hemoglobin HGB/ RBC x
these young RBCs
● expresses the mass 10
of hemoglobin per cell ● Vital (or “supravital”) stains are dyes
and parallels the absorbed by live cells
MCHC (pictogram/pg) ➔ Young RBCs contain ribonucleic acid (RNA) and
are called reticulocytes when the RNA is visualized
MCHC Mean corpuscular hemoglobin HGB/ HCT x using vital stains
concentration 100 WHITE BLOOD CELLS
● reflects RBC staining ➔ are a loosely related category of cell types
intensity and amount
dedicated to protecting their host from infection and
of central pallor.
(g/dL) injury
➔ WBCs are transported in the blood from their
RDW Red blood cell Distribution source, usually bone marrow or lymphoid tissue, to
width their tissue or body cavity destination.
● Expresses the degree ➔ WBCs may be counted visually using a microscope
of variation in RBC and hemacytometer
volume.
➔ the typical dilution is 1:20, and the diluent is a dilute
● Based on the
standard deviation of acid solution.
RBC volume ● The acid causes RBCs to lyse or rupture;
● Routinely reported by without it, RBCs, which are 500 to 1000
automated blood cell times more numerous than WBCs, would
analyzers obscure the WBCs.
➔ Medical laboratory professionals who analyze body
BLOOD FILM EXAMINATION
fluids such as cerebrospinal fluid or pleural fluid
OBJECTIVE : Oil Immersion
may employ visual WBC counting
Feathery edge – cells are uniformly distributed
➔ A decreased WBC count is called leukopenia
● To accomplish a blood film examination, the
➔ An increased WBC count is called leukocytosis
microscopist prepares a “wedge-prep” blood film
on a glass microscope slide, allows it to dry, and
fixes and stains it using Wright or Wright-Giemsa
stain.
● The microscopist visually performs an estimate of
the WBC Count
○ 40x or 50x obj. at 400x or 500x
magnification
○ Platelet count with 100x oil immersion
objective at 1000x magnification
○ For comparison with their respective
analyzer counts, and investigates
discrepancies.
● The microscopist systematically reviews, identifies,
and tabulates 100 (or more) WBCs to determine
their percent distribution
○ This process is referred to as WBC
differential (“diff”)
○ Relies on the microscopist’s skill, visual
acuity and integrity, and provides extensive
GRANULOCYTES
diagnostic information.
● The microscopist examines the morphology of
NEUTROPHILS
WBCS, RBCs, and platelets by light microscopy for
abnormalities of shape, diameter, color, or inclusion ➔ Phagocytic cells whose
using 1000x magnification. major purpose is to engulf
and destroy
RETICULOCYTES microorganisms and

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 foreign material, either directly or after they have
been labeled for destruction by the immune system
➔ The term segmented refers to their multilobed
nuclei.
➔ The cytoplasm of neutrophils contains pink- or
lavender-staining granules filled with bactericidal
substances.
➔ An increase in neutrophils is called neutrophilia
● signals bacterial infection.
➔ A decrease is called neutropenia
● often caused by certain medications or
viral infections.
BANDS
➔ are slightly less mature
neutrophils with a nonsegmented
nucleus in a U or S shape
➔ An increase in bands also signals
bacterial infection and is
customarily called a left shift.
EOSINOPHILS
➔ are cells with round, bright
orange-red cytoplasmic granules
filled with proteins involved in
immune system regulation.
➔ An elevated eosinophil count is
called eosinophilia
● often signals a response
to allergy or parasitic infection.
BASOPHILS
➔ are cells with dark purple,
irregular cytoplasmic granules
that obscure the nucleus.
➔ The basophil granules contain
histamines and various other
proteins.
➔ An elevated basophil count is called basophilia.
● Basophilia is rare and often signals a
hematologic disease.
AGRANULOCYTES
LYMPHOCYTES
➔ comprise a complex system of
cells that provide for host
immunity.
➔ On a Wright-stained blood film,
most lymphocytes are nearly
round, are slightly larger than
RBCs, and have round featureless
nuclei and a thin rim of nongranular cytoplasm.
➔ An increase in the lymphocyte count is called
lymphocytosis
● often is associated with viral infections
➔ abnormally low lymphocyte count is called
lymphopenia or lymphocytopenia
● often associated with drug therapy or
immunodeficiency.
MONOCYTES
➔ is an immature macrophage
passing through the blood from its
point of origin, usually the bone
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marrow, to a targeted tissue location.
➔ are the most abundant cell type in the body
although monocytes comprise a minor component
of peripheral blood WBCs.
➔ Macrophages occupy every body cavity; some are
motile and some are immobilized.
➔ Their tasks are to identify and phagocytize (engulf
and consume) foreign particles and assist the
lymphocytes in mounting an immune response
through the assembly and presentation of antigen
epitopes
➔ An increase in the number of monocytes is called
monocytosis
● Benign monocytosis may be found in
certain infections or in inflammation
➔ a decreased monocyte count, so the theoretical
term monocytopenia is seldom used.
PLATELETS
➔ are only 2 to 4 mm in diameter,
round or oval, anucleate
➔ are true blood cells that maintain
blood vessel integrity by
initiating vessel wall repairs
➔ Platelets rapidly adhere to the
surfaces of damaged blood
vessels, form aggregates with neighboring platelets
to plug the vessels, and secrete proteins and small
molecules that trigger thrombosis, or clot
formation.
➔ are the major cells that control hemostasis, a series
of cellular and plasma-based mechanisms that seal
wounds, repair vessel walls, and maintain vascular
patency (unimpeded blood flow)
➔ Uncontrolled platelet and hemostatic activation is
responsible for deep vein thrombosis, pulmonary
emboli, acute myocardial infarctions (heart attacks),
cerebrovascular accidents (strokes), peripheral
artery disease, and repeated spontaneous
abortions (miscarriages)
➔ The microscopist counts platelets using the same
technique used in counting WBCs on a
hemacytometer, although a different counting area,
diluent, and dilution is usually used
➔ Elevated platelet counts, called thrombocytosis
● signal inflammation or trauma but convey
modest intrinsic significance.
➔ Essential thrombocythemia is a rare malignant
condition characterized by extremely high platelet
counts and uncontrolled platelet production
➔ A low platelet count, called thrombocytopenia
● a common consequence of drug treatment
and may be life-threatening.
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 be performed behind a barrier, such as a plastic
shield, or protective eyewear should be worn. .
7. Personal protective clothing and equipment must be
provided to the laboratory staff. The most common
forms of personal protective equipment are
described in the following section:
❖ a. Lab gowns
❖ b. Gloves must be worn when the potential
for contact with blood or body fluids exists)
and when venipuncture or skin puncture is
performed.
❖ c. Eyewear when there is potential for
aerosol mists, splashes, or sprays to
SAFETY IN THE HEMATOLOGY LABORATORY mucous membranes (mouth, eyes, or
nose).
SAFETY OFFICER 8. Phlebotomy trays should be appropriately labeled to
➔ a designated safety officer is a critical part of a indicate potentially infectious materials.
laboratory safety program. This individual has many 9. If a pneumatic tube system is used to transport
duties affecting staff including compliance with specimens, the specimens should be transported in
existing regulations affecting the laboratory and the appropriate tube (primary containment), and
staff placed into a special self-sealing leakproof bag
ORGANIZATIONS appropriately labeled with the biohazard symbol
➔ U.S. Department of labor’s Occupational Safety and (secondary containment).
Health Administration. (OSHA) 10. When equipment used to process specimens
➔ Clinical and laboratory standards institute becomes visibly contaminated or requires
➔ CDC, part of the U.S. department of health, maintenance or service, it must be decontaminated,
Department of Health and Human Services whether the service is performed within the
(DHHS), Public health council (?) laboratory or by a manufacturer repair service.
➔ College of American Pathologists ( CAP) OCCUPATIONAL HAZARDS
➔ The joint commission (the joint commission on ➔ four important occupational hazards in the
accreditation of healthcare organizations) laboratory: fire hazard, electrical hazard, chemical,
STANDARD PRECAUTIONS and needle puncture
➔ One of the greatest risks associated with the
hematology laboratory is the exposure to blood and
body fluids
➔ In December 1991, the OSHA issued a final rule for
the occupational exposure to bloodborne pathogens
standard, universal precautions, original term;
OSHA’s current terminology is standard
precautions.
➔ the term standard precautions is used to describe
— all blood, body fluids, and tissues that are to be
handled as though they are potentially infectious
APPLICABLE SAFETY PRACTICES REQUIRED BY THE
OSHA STANDARD
1. Eating, drinking, smoking, and applying cosmetics
or lip balm must be prohibited in the laboratory work
area.
2. Hands, pens, and other fomites must be kept away
from the mouth and all mucous membranes.
3. Mouth pipetting must be prohibited.
4. Needles and other sharp objects contaminated with
blood and other potentially infectious materials
should not be manipulated in any way.
5. .Contaminated sharps must be placed in a
puncture-resistant container that is appropriately
labeled with the universal biohazard symbol or a red
container that adheres to the standard. The
container must be leakproof.
6. Removing caps when checking for clots, filling
hemacytometer chambers, making slides,
discarding specimens, making dilutions, and
pouring specimens or fluids- These procedures may

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