Chapter 1: An Overview of Clinical Laboratory Hematology

Blood- transports oxygen from lungs to tissue

clears tissues of carbon dioxide
transports glucose, proteins, and fats
Moves wastes to the liver and kidneys.
Plasma
The liquid portion of the blood among many other
components
coagulation enzymes that protect vessels from trauma and
maintain the circulation
Transports and nourishes blood cells.
Three families of blood cells
1. red blood cells (RBCs) or erythrocytes
2. white blood cells (WBCs) leukocytes
3. platelets or thrombocytes

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Buffy Coat
light-colored layer between the RBCs and plasma
Contains WBCs and platelets.

Three Numerical Results

1. RBC count
2. Hemoglobin
3. Hematocrit

To compute the RBC indices

1. mean cell volume (MCV)
2. mean cell hemoglobin (MCH)
3. mean cell hemoglobin concentration

Mean Cell Volume (MCV)

measure of volume
reflects RBC diameter on a Wright-stained blood film
Mean Cell Hemoglobin Concentration (MCHC)
reflects RBC staining intensity or degree of pallor
Mean Cell Hemoglobin (MCH)
expresses the mass of hemoglobin and closely reflects the
MCHC
RBC distribution width (RDW)
expresses the degree of variation in RBC volume
Extreme RBC volume variability is visible on the Wrightstained blood film as variation in diameter and is called
anisocytosis.
based on the standard deviation of RBC volume
Reported by automated cell counters but cannot be provided
using manual RBC measurements.

Hematology- the study of blood cells

Wrights Romanowsky- type stain (polychromatic, a mixture of acidic

and basic dyes) remains the heart of blood cell identification
Red Blood Cells

Red Blood Cells (RBC)

a nucleate biconcave cells filled with a reddish protein,
hemoglobin (Hb, HGB) which trans- ports oxygen and
carbon dioxide
pink to red ; 6 to 8 m
Anemia loss of oxygen-carrying capacity; reduced RBC count
Polycythemia increased RBC count reflecting increased body
RBC mass; leads to hyperviscosity

To count RBCs, laboratory scientists carefully pipetted a tiny

aliquot of whole blood and mixed it with 0.85% (normal)
saline.
A 1 : 200 dilution was typical for RBC counts, and a glass
pipette designed to provide this dilution

Thoma pipette used routinely until the advent of automation

The diluted blood was transferred to a counting
chamber or
Report the RBC count in cells per microliter (mcL),
milliliter (mL, also called cubic centimeter, or cc), or liter
(L).
Coulter Counters first electronic counter by Joseph and
Wallace Coulter of Chicago, Illinois, was used so widely that
today automated cell counters
Hemoglobin measurement relies on a weak solution of
potassium cyanide and potassium ferricyanide, called
Drabkin reagent.
Hemoglobin
is
converted
to
stable
cyanmethemoglobin(hemiglobincyanide), and the solution is
placed in a photometer at 540 nm wavelength.
Some automated hematology profiling instruments use a
formulation
of
the
ionic
surfactant
(detergent) sodium
dodecyl
sulfate to reduce
environmental cyanide.
Hematocrit
ratio of the volume of RBCs to the volume of whole blood
determined
by
transferring
blood
to
a
graduated plastic tube, centrifuging, measuring the column
of
RBCs, and dividing by the total length of RBCs plus
plasma
The normal ratio approaches 50%
Also called packed cell volume (PCV), the packed cells
referring to RBCs.

RBC count, hemoglobin, hematocrit, indices, and RBC

morphology are used to detect, diagnose, assess the severity
of, and monitor the treatment of anemia, polycythemia, and
numerous systemic conditions that affect RBCs

Reticulocytes
Polychromatophilic erythrocytes- in the Wright-stained film,
1% to 2% of RBCs exceed the 6- to 8-m ; slightly blue-gray
newly released from the RBC production site, the bone
marrow
closely observed because they indicate bone marrow
regeneration during blood loss and certain anemias
Methylene blue dyes- called nucleic acid stains or vital stain
used to differentiate and count these young RBCs
Vital stains -dyes absorbed by live cells. Young RBCs
contain ribonucleic acid (RNA) and are called reticulocytes
when the RNA is highlighted using vital stains.

White Blood Cells

White Blood Cells or Leukocytes (WBC)

are not really blood cells
they are a loosely related grouping of cell families
dedicated to protecting their host from infection and injury
WBCs hitch a ride in the blood from their source,
usually bone marrow or lymphoid tissue, to their tissue
destination.
They are so named because they are nearly colorless in
an unstained cell suspension
WBCs may be counted visually using a microscope, hemacytometer, and a Thoma pipette
The technique is the same as RBC counting, but the typical
dilution is 1: 20, and the diluent is composed of dilute acetic
acid in normal saline.

Chronic leukemia
an extreme increase in the WBC count imparts a milky
appearance to the blood

Chapter 1: An Overview of Clinical Laboratory Hematology

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**Acid causes RBCs to lyse or rupture

Leukopenia
decreased WBC count (fewer than 4500/mcL)
Leukocytosis
increased WBC count (more than 11,500/ mcL)

Types of White Blood Cells

1. Polymorphonuclear neutrophils (PMNs, or segmented
neutrophils or Segs
phagocytic cells whose sole purpose is to engulf and
destroy bacteria that have been earlier labeled as
harmful by the immune system.

Neutrophilia
increase in segs; signals bacterial infection.
Neutropenia
decrease in segs; caused by long-term drug administration
or a viral infection.

2.

3.
-

Band neutrophils (bands)

part of the seg family
less differentiated or less mature than segs
increase also signals bacterial infection and is customarily
called a left shift
The cytoplasm
of segmented neutrophils and bands
contains submicroscopic, pink-staining granules filled with
bactericidal secretions.
Eosinophils (Eos)
bright orange, regular cytoplasmic granules filled with
antihistamine.

Eosinophilia
elevated eosinophil count is called and
response to allergy or parasitic infection.
4.
-

signals

most common form of childhood leukemia

6.
-

Monocytes (monos)
immature macrophage passing through the blood from its
point of origin, usually the bone marrow, to a targeted
tissue location

Macrophages
Most abundant cell in the body, more abundant than RBCs
or skin cells, although they are a minor component of the
blood film differential count
Some are motile and some immobilized.
Their task is to identify and phagocytose (engulf) foreign
particles and assist the lymphocytes in mounting an
immune response through the assembly and presentation
of immunogenic epitopes
Monocytosis
An increase in the number of monocytes may signal a
hematologic disease, such as leukemia

Platelets

Basophils (basos)
dark purple, irregular cytoplasmic granules that obscure
the nucleus

Basophilia
elevated basophil count
signals a hematologic disease, such as leukemia.

Platelets or thrombocytes
Are true blood cells that maintain blood vessel integrity
by instigating vessel wall repairs
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. Platelets are the cells
that control hemostasis,
Hemostasis
Series of cellular and plasma-based mechanisms that seals
wounds, repairs vessel walls, and maintains vascular
patency.
Thrombocytosis
Elevated platelet counts; signal inflammation or trauma but
carry small intrinsic significance.
Essential thrombocythemia
rare malignant condition characterized by extremely high
platelet counts and uncontrolled platelet production.
Thrombocytopenia
low platelet count
a common consequence of drug treatment and may be
life-threatening
accounts
for
the
majority of hemorrhage-related
emergency department visits.

** Segs, bands, eosinophils, and

basophils- collectively called
granulocytes because of their prominent cytoplasmic granules

Leukemia
is uncontrolled proliferation of WBCs.
may be chronic,
for example chronic myelogenous
(granulocytic) leukemia, or acute,
such as
acute
myeloblastic leukemia
5.
-

Lymphocytes (lymphs)
Complex system of cells that provide for host immunity
recognize
foreign
antigens
and mount antibody
(humoral) and cell-mediated antagonistic responses

Lymphocytosis
Increase in the lymphocyte count; associated with viral
infections.
Lymphopenia or lymphocytopenia
abnormally low lymphocyte count is and is associated with
long-term drug therapy or immunodefi- ciency
Chronic lymphocytic leukemia
prevalent in people older than 70 years
Acute lymphoblastic leukemia

Complete Blood Count

Complete Blood Count

The scientist is responsible for the integrity of the specimen
and ensures that it is free of clots, hemolysis, and
inappropriate anticoagulant- to-specimen ratios known as
short draws.
When one of the results from the profiling instru- ment is
abnormal, the instrument provides an indication of this,
sometimes called a flag. In this case, the scientist performs
a reflex blood film examination

Blood Film Examination

Chapter 1: An Overview of Clinical Laboratory Hematology

Blood Film Examination

wedge-prep blood film on a glass microscope slide,
allows it to dry, and fixes and stains it using Wright or
Wright-Giemsa stain
Endothelial cells
Important in maintaining normal blood flow, in snaring
platelets during times of injury, and in enabling WBCs to
escape from the vessel to the surrounding tissue when
called upon.
Plasma coagulation
is one component of hemostasis; another is platelets
Coagulation system
complex sequence of plasma proteins, some enzymes,
and some enzyme cofactors to produce clot formation
after blood vessel injury.
Erythroid series
precursors to RBCs

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Myeloid series cells

mature to form bands and segmented neu- trophils,
eosinophils, and basophils
Megakaryocytes
produce platelets
Osmotic fragility test
Uses graduated concentrations of saline
solutions to
detect spherocytes, RBCs with proportionally reduced
surface membrane area, in hereditary spherocytic or warm
autoim- mune hemolytic anemia
Glucose-6-phosphate dehydrogenase assay tests
For an inherited RBC enzyme deficiency causing severe
episodic hemolytic anemia
Hb electrophoresis

used to detect and diagnose sickle cell anemia and

other inherited qualitative hemoglobin abnormalities and
thalassemias