OVERVIEW OF HEMATOLOGY

Nelven M. Gallego, RMT, MLS(ASCPi), MSMT cand.

Far Eastern University, Manila
Institute of Arts and Sciences (IAS)
Department of Medical Technology
Hematology 1 Lecture (MTY1211)
Nelven M. Gallego, RMT, MLS(ASCPi)
AN OVERVIEW
HEMATOLOGY:  FUNCTIONS OF BLOOD:
➢ comes from two Greek (1) Respiratory
words Haima and Logos (2) Nutritional
(3) Excretory
➢ it is the discipline that studies the development
and diseases of blood (4) Buffering action
(5) Maintenance of body temp
(6) Transport of hormones
(7) Defense mechanism
BLOOD:
 Average human: 5 liters

Nelven M. Gallego, RMT, MLS(ASCPi)

BLOOD COMPOSITION
A. LIQUID PORTION:  CHARACTERISTICS:
(1) Serum (2) Plasma (1) Fluid (2) Red
(3) Slightly alkaline
B. SOLID PORTION/ (4) Average S.G. of 1.055

THREE CATEGORIES: (5) Thick and viscous

(1) Erythrocytes (RBC) (6) Makes up 75-85 mL per kilogram of body weight

(7) About 20 grams of solids per 100 mL of blood

(2) Leukocytes (WBC)
(3) Thrombocytes (PLT)
C. GASEOUS PORTION:

Nelven M. Gallego, RMT, MLS(ASCPi)

BLOOD COMPOSITION
PLASMA:
➢ Liquid portion that transports
and nourishes blood cells
 Provides coagulation enzymes that protect the blood vessels

BUFFY COAT:
➢ Light-layer between the
RBC and plasma
 Contains WBCs and PLATELETS
 Excluded from the hematocrit detrmination

Nelven M. Gallego, RMT, MLS(ASCPi)

BLOOD COMPOSITION
 GRANULOCYTE:
(1) Basophil
(2) Eosinophil
(3) Neutrophil
(4) Band
 AGRANULOCYTE:
(1) Lymphocyte
(2) Monocyte

Nelven M. Gallego, RMT, MLS(ASCPi)

RED BLOOD CELLS
ERYTHROCYTES:  PERIPHERAL BLOOD FILM:
➢ anucleate, biconcave, (1) pink to red
discoid cells filled (2) 6-8 um in diameter
with hemoglobin (3) central pallor (1/3rd)
(4) biconcave
HEMOGLOBIN
 a reddish protein that transports O2 and CO2

Nelven M. Gallego, RMT, MLS(ASCPi)

RED BLOOD CELLS
ANEMIA: PARAMETERS:
➢ Means loss of oxygen-
(1) RBC count
carrying capacity
(2) Hemoglobin
 🔽 RBC
(3) Hematocrit
 count
🔽 RBC HGB (4) RBC
concentration
(5) indices
RBC morphology
POLYCYTHEMIA:
➢ Increased circ. RBC mass
 🔼 RBC count —>
hyperviscosity

Nelven M. Gallego, RMT, MLS(ASCPi)

RED BLOOD CELL COUNT
THOMA PIPETTE:
1:200 DILUTION:
➢ a glass pipette that was
used routinely until the  typical for RBC count
advent of automation
0.85% NSS:
 matches the osmolality of the blood
 neither swelling nor shrinking

Nelven M. Gallego, RMT, MLS(ASCPi)

RED BLOOD CELL COUNT
HEMACYTOMETER:
➢ a glass counting chamber;
aka hemocytometer
REPORTING:
 RBC count is reported in cells per microliter
(mL, mcL, also called cubic millimeter, mm^3),
milliliter (mL, also called cubic centimeter, or
cc), or liter (L)

Nelven M. Gallego, RMT, MLS(ASCPi)

RED BLOOD CELL COUNT
VISUAL RBC COUNTING:
➢ Developed before 1900
and although inaccurate,
was the only way to
count RBCs until 1958

COULTER COUNTER:
 First electronic counter, patented in 1953 by Joseph
and Wallace Coulter of Chicago, Illinois

 Coulter principle of direct current electrical impedance

Nelven M. Gallego, RMT, MLS(ASCPi)

HEMOGLOBIN DETERMINATION
DRABKIN REAGENT:
➢ Potassium cyanide and potassium ferricyanide

CYANMETHEMOGLOBIN:
 Hemoglobincyanide

SODIUM LAURYL SULFATE:

 Ionic surfactant; reduce environmental cyanide

Nelven M. Gallego, RMT, MLS(ASCPi)

HEMATOCRIT DETERMINATION
MICROHEMATOCRIT:
➢ HCT is the ratio of the
volume of packed RBCs to
the volume of whole blood
 PACKED CELL VOLUME (PCV)

 In percentage (%) or in liters per liter (L/L)

 Normal ratio: 50%

*no clot activator, it takes 10 mins to clot

Nelven M. Gallego, RMT, MLS(ASCPi)

RED BLOOD CELL INDICES

80-100 fL

26-34 pg

32-36 g/dL

Nelven M. Gallego, RMT, MLS(ASCPi)

RED BLOOD CELL INDICES
MEAN CELL VOLUME:
➢ a volume measurement recorded
in femtoliters (fL)

MEAN CELL HEMOGLOBIN CONC:

➢ expressed in g/dL

Nelven M. Gallego, RMT, MLS(ASCPi)

RED BLOOD CELL INDICES
MEAN CELL HEMOGLOBIN:
➢ expressed in picograms (pg)

RBC DISTRIBUTION WIDTH:



Nelven M. Gallego, RMT, MLS(ASCPi)

RED BLOOD CELL MORPHOLOGY
ANISOCYTOSIS: POIKILOCYTOSIS:
➢ general term used in ➢ general term for mature erythrocytes that
have a shape other than the normal round,
hematology to denote
biconcave appearance
an increased variation
in cell size

Nelven M. Gallego, RMT, MLS(ASCPi)

RBC MORPHOLOGY
LIGHT MICROSCOPY:
 RED BLOOD CELL MORPHOLOGY
a. RBC diameter
b. Color or hemoglobinization
c. RBC shape
d. Cytoplasmic inclusions

Nelven M. Gallego, RMT, MLS(ASCPi)

RETICULOCYTES
RETICLOCYTES:
➢ Newly released from the
RBC production site: the
bone marrow
 0.5% to 2% of RBCs exceeds 6- to 8-mm
average diameter

 Stain slightly blue-gray

 those are polychromatic (polychromatophilic)

erythrocytes

Nelven M. Gallego, RMT, MLS(ASCPi)

RETICULOCYTES
METHELYNE BLUE DYES:
➢ Nucleic acid stains or
vital stains, are used to
differentiate and count
these young RBCs
 Vital (or “supravital”) stains are dyes absorbed
by live cells

 Young RBCs contain ribonucleic acid (RNA)

Nelven M. Gallego, RMT, MLS(ASCPi)

RETICULOCYTE COUNT

Nelven M. Gallego, RMT, MLS(ASCPi)

REFERENCE INTERVALS (ADULT)

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
LEUKOCYTES:
➢ Loosely related category
of cell types dedicated to
protecting their host from
infection and injury
 WBCs are transported in the blood from bone
marrow or Lymphoid tissue

 Nearly colorless in an unstained cell suspension

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELL COUNT
WBC COUNT:
➢ The technique is the same 1:20 DILUTION:
as RBC counting  typical for WBC count

DILUENT:
 a dilute acid solution

 it causes RBCs to Lyse or rupture

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
LEUKOCYTOSIS: LEUKOPENIA:
➢ An increased WBC count ➢
(more than 11,500/uL)

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
NEUTROPHILS:
➢ A phagocytic cells whose
major purpose is to engulf
and destroy microorganisms
and foreign material

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
NEUTROPHILIA: NEUTROPENIA:
➢ An increased in ➢ A decreased in neutrophils
neutrophils

 often signals bacterial infection

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
BANDS:
➢ Bands are less differentiated
or less mature neutrophils
 Pink- or lavender-staining granules filled with
bactericidal secretions

 LEFT SHIFT: an increase in

bands; signals bacterial
infection

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
EOSINOPHILS:
➢ are bright orange-red, regular cytoplasmic granules filled
with proteins involved in immune system regulation

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
EOSINOPHILIA: EOSINOPENIA:
➢ An elevated eosinophils ➢ theoretical and not used
 often caused signals a response in allergy or
parasitic infection

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
BASOPHILS:
➢ Basophils are cells with
dark purple, irregular
cytoplasmic granules that
obscure the nucleus

 Basophil granules contain histamines and various

other proteins

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
BASOPHILIA: BASOPENIA:
➢ An elevated basophils ➢ theoretical and not used
 often signals hematologic diseases

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
LYMPHOCYTES:
➢ Lymphocytes recognize foreign
antigens and mount humoral
(antibodies) and cell-mediated
antagonistic responses.

 Most lymphocytes are nearly round, are slightly larger

than RBCs, and have round featureless nuclei and
a thin rim of nongranular cytoplasm

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
LYMPHOCYTOSIS: LYMPHOPENIA:
➢ An increased lymphocyte ➢ Lymphocytopenia
 often is associated with viral infections
 often associated with drug therapy or
immunodeficiency
 accompanying lymphocytosis are often
variant or relative lymphocytes

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
LEUKEMIA: ALL:
➢ An uncontrolled ▪ most common form of childhood leukemia
proliferation of WBCs

CLL:
 more prevalent in people older than 65 years

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
MONOCYTES:
➢ An immature macrophage  assist the lymphocytes in mounting an immune
response through the assembly and presentation
passing through the blood of immunogenic epitopes
from its point of origin
 Macrophages comprise minor component of
peripheral blood WBC

 Their tasks are to identify and phagocytose

(engulf and consume) foreign particles

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
MONOCYTES:
➢ Monocytes have a
slightly larger diameter
than other WBCs, blue-
gray cytoplasm with fine
azure granules, and a
nucleus that is usually
indented or folded.

Nelven M. Gallego, RMT, MLS(ASCPi)

WHITE BLOOD CELLS
MONOCYTOSIS MONOCYTOPENIA:
➢ An increase in the ➢ Seldom used
number of monocytes
 Found in certain infections, collagen-vascular diseases,
or in acute and chronic leukemias

Nelven M. Gallego, RMT, MLS(ASCPi)

THROBOCYTES
PLATELETS:
➢ True blood cells that
HEMOSTASIS:
maintain blood vessel  a series of cellular and plasma based mechanism
that seal wounds, repair vessel walls, and maintain
integrity by initiating vascular patency
vessel wall repairs
 Secrete proteins and small molecules that trigger
thrombosis, or clot formation

Nelven M. Gallego, RMT, MLS(ASCPi)

THROBOCYTES
PLATELETS:
 Platelets are only 2 to
4 mm in diameter, round
or oval, anucleate and
slightly granular

 Some hematologists prefer to call platelets

as “cell fragments”

Nelven M. Gallego, RMT, MLS(ASCPi)

THROBOCYTES
THROMBOCYTOSIS: THROMBOCYTOPENIA:
➢ elevated platelet count ➢ a low platelet count
 signal inflammation trauma but convey modest
 a common consequence of drug treatment and may
intrinsic significance be life-threatening

Nelven M. Gallego, RMT, MLS(ASCPi)

THROBOCYTES
ESSENTIAL UNCONTROLLED PLATELET &
THROMBOCYTEMIA: HEMOSTATIC ACTIVATION:
a. Deep vein thrombosis
➢ a rare malignant
b. Pulmonary emboli
condition characterized
c. Acute myocardial infarctions
by extremely high
d. Cerebrovascular accidents
platelet counts and
e. Peripheral artery disease
uncontrolled platelet
f. Repeated spontaneous abortions
production

Nelven M. Gallego, RMT, MLS(ASCPi)

COMPLETE BLOOD COUNT
COMPLETE BLOOD COUNT:
➢ Performed on automated
hematology profiling
instruments and includes
the RBC, WBC, and
platelet measurements

SPECIMEN:
 Appropriate anticoagulant and tube, free from clots
and hemolysis, sufficient volume, tested and prepared
for storage within the appropriate time frame

Nelven M. Gallego, RMT, MLS(ASCPi)

COMPLETE BLOOD COUNT
SPECIMEN ACCESION:
 Accurately registered in
the work list; e.g. barcode
or radio frequency
identification technology
FLAG:
➢ An indication when one
of the results from the
profiling is abnormal
 A “reflex” blood film
Nelven M. Gallego, RMT, MLS(ASCPi)
BLOOD FILM EXAMINATION
WRIGHT-GIEMSA STAIN:
 RBC and Platelets: shape,
diameter, color, or inclusion
 Oil immersion Lens: 50x
or 100x —> 500x or 1000x

WBC DIFFIERENTIAL:
 A review and tabulation of 100 to 200 white
blood cells (WBCs) in a stained blood film

Nelven M. Gallego, RMT, MLS(ASCPi)

REFERENCE INTERVALS (ADULT)

Nelven M. Gallego, RMT, MLS(ASCPi)

ENDOTHELIAL CELLS
ENDOTHELIAL CELLS:
 Form the inner surface
of the blood vessel
Structural and do not flow in the bloodstream


Seldom studied in the hematology laboratory



Nelven M. Gallego, RMT, MLS(ASCPi)

COAGULATION
PRIMARY HEMOSTASIS: COAGULATION TESTS:
 It refers to the role of (1) Platelet count
blood vessels and (2) Mean platelet volume
platelets in response to (3) Prothrombin time
a vascular injury. (4) APTT
(5) Thrombin time
SECONDARY HEMOSTASIS: (6) Fibrinogen assay
 Describes the activation of a series of coagulation (5) D-dimer assay
proteins in the plasma

Nelven M. Gallego, RMT, MLS(ASCPi)

COAGULATION FACTORS

Nelven M. Gallego, RMT, MLS(ASCPi)

COAGULATION
FIBRINOLYSIS:
➢ Final stage of coagulation

 a third system of enzymes and cofactors digests

clots to restore vessel patency

ASSIGNED READINGS:
a. Advance Hematology Procedures
b. Additional Hematology Procedures
c. Hematology QA and QC

Nelven M. Gallego, RMT, MLS(ASCPi)

MEDICAL TERMINOLOGY

Nelven M. Gallego, RMT, MLS(ASCPi)

MEDICAL TERMINOLOGY

Nelven M. Gallego, RMT, MLS(ASCPi)

OVERVIEW OF HEMATOLOGY

Nelven M. Gallego, RMT, MLS(ASCPi), MSMT cand.

Far Eastern University, Manila
Institute of Arts and Sciences (IAS)
Department of Medical Technology
Hematology 1 Lecture (MTY1211)
Nelven M. Gallego, RMT, MLS(ASCPi)