Basic Hematology

Clinical Laboratory Testing

Basic Hematology
Hematology section
Study of formed elements of blood and
blood forming tissues
Screen, diagnosis and monitor hematologic
diseases (anemias, leukemias, etc.) and also
give a general idea of overall health
 History
Hippocrates (400 B.C.) : describes
blood as composite of 4 humours
Athanasius Kircher (1657): worms in
blood
Anton van Leeuwenhoek (1674):
account of RBCs
Giulio Bizzozero (late 1800s) :
platelet as petites plaque
 History
Karl Vierordt (1852) : published the 1st
quantitative results of blood cell analysis
James Homer (1902): develop Wrights
stain
Improvements in the methods of blood
examination
Modern hematologists: alterations in the
components of blood is a result of disease,
not primary cause of it.
 Composition of Blood - blood is 6-8% of total
body weight and equals approximately 5 liters

Serum vs Plasma
Both are fluid portions of blood
Serum is from clotted blood, has no clotting factors
(used up in the clotting process)
Plasma is from anticoagulated blood, has clotting
factors, makes up about 55% of bloods volume
COMPOSITION
 PLASMA
Unclotted
portion
Pale Yellow fluid
Water
Proteins
Other
substances
Plasma proteins
ALBUMIN
GLOBULIN
FIBRINOGEN
 ALBUMIN

58% of proteins

Contributes to the osmotic pressure

The water balance between blood and tissues is
determined by the movement of water into and out of
the blood by osmosis.

Also functions as transport molecules for hormones

 GLOBULIN
38% of proteins

Some globulins, such as antibodies and

complement, are part of the immune
system

Also functions as transport molecules for

hormones

Helps in the formation of blood clot

 FIBRINOGEN

CLOTTING FACTOR
4% of plasma proteins
Converted into fibrin during Activation of
clotting factors

Serum- plasma without clotting factors

Clotted blood
 FORMED ELEMENTS
RED BLOOD CELLS
WHITE BLOOD CELLS
Granulocytes
Neutrophil
Basophil
Eosinophil

Agranulocytes
Lymphocyte and Monocyte

PLATELET OR
THROMBOCYTES
 Erythrocytes / RBCs
6-8 micrometer (diameter), biconcave disc
non-nucleated , with hemoglobin
transport oxygen and carbon dioxide
~ 4-6 million/L : RBC Count
how to count: thru manual or automated

Anemia vs Polycythemia

Electron Micrograph Stained Blood Smear - Light Microscope

 Hemoglobin, Hematocrit and
RBC indices
Hgb

Hgb determination uses Drabkins

Reagent
Automated methods uses Sodium
dodecyl sulfate
 Hemoglobin, Hematocrit and
RBC indices
Hct
Hct : ratio of the volume of RBCs to
volume of whole blood
PCV

Normal ratio = 50% (estimated)

 Hemoglobin, Hematocrit and
RBC indices
RBC indices
computed based on RBC ct, Hct and
Hgb
MCV: a measure of volume; RBC
diameter
MCHC: reflects RBC staining intensity
or degree of pallor
MCH : expresses mass of hgb

RDW : degree of anisocytosis

 RBC index Formula Reference
Range

MCV (Hct / RBC ct) X 10 80-100 fL

MCH (Hgb/RBC ct) X 10 26-32 pg

MCHC (Hgb/Hct) X 100 32-36% (g/dL

in SI)

RDW (Std Dev of 11.5-14.5%

MCV/MCV) X 100
 Reticulocyte
Polychromatophilic RBCs
About 1-2%
Newly released RBCs from the BM

Contains RNA

To count:

Supravital dye
Automation (TOA/Sysmex)
 Leukocytes / WBCs
Hitch a ride in the blood
Colorless in unstained smears
~ 4,500-11,500 /L in circulating blood; many
more in tissues
Function in immunity and defense against
bacteria, viruses and all other foreign
materials (includes allergens like pollen, etc.)
Leucopenia vs Leucocytosis
Differentiated using Wrights stain
 Types of WBC
According to nucleus
-PMN
-Mononuclear

Accordingto cytoplasmic granules

-Granulocytes
-Agranulocytes
 Types of WBCs:
Granulocytes (neutrophils, basophils, &
eosinophils)

Drawing Stained Granulocytes Viewed Under

Microscope
 Agranulocytes

Lymphocytes (On blood smear)

Monocytes (On blood smear)

 PMN/Segmented neutrophils
Neutrophilia : bacterial infection
Neutropenia : long term drug
administration; viral infection
Bands

Shift to the left

 Eosinophils

With bright orange granules filled

with antihistamine
Eosinophilia : allergy; parasitism

Eosinopenia: theoretical/ unused

 Basophils

with dark blue granules that obscure

the nucleus
Basophilia : rare, hematologic
disease (leukemia)
Basopenia : theoretical/unused
 Lymphocytes
B, T and NK cells
Responsible for CMI and HI

Big round nucleus, thin rim of

cytoplasm
Slightly larger then RBCs

Lymphocytosis : viral infection

Lymphopenia/Lymphocytopenia :
long term drug administration;
immunodeficiency
 Monocytes
Macrophage in the tissues
Phagocytosis, presentation of
epitopes
Slightly larger than other WBCs

Monocytosis : hematologic disorder

Monocytopenia : theoretical
Platelets / Thrombocytes
Pieces of a bone marrow cell known as a
megakaryocyte
Function to stop bleeding by forming a plug
and also release coagulation factors
(controls hemostasis)
Forms thrombus (clot)
Capable of adhesion, aggregation, secretion
2-4 , oval, anucleated, slightly granular
 Platelets / Thrombocytes
Reference Range: 150-450 K/cu.mm.
Counted using manual or automated

Automated: with MPV (6.8-10.2%)

High MPV: regenerative BM response

to platelet consumption
Thrombocytosis : inflammation;
trauma
Thrombocytopenia : consequence of
drug treatment
Common Hematology Laboratory
Tests
*check the integrity of the specimen!!!!!
-short draws
Complete Blood Count (CBC) includes:
RBCcount
WBC count
Hemoglobin
Hematocrit
WBC Differential count

Automated: flags (when one of the results is

abnormal) back up procedure
 Common Hematology
Laboratory Tests
Plateletcount
Additional information on size and appearance
of blood cells (blood film examination)
 Common Hematology Laboratory Tests
Erythrocyte Sedimentation Rate (ESR)
Reticulocyte Count
Peripheral Blood Examination
Sickle Cell Testing
Osmotic Fragility Test
Coagulation Tests include:
Prothrombin Time (PT)
Partial Thromboplastin Time (PTT)

Fibrinogen

Factor Analysis
 ADVANCED HEMA
PROCEDURES
BM Examination
Cytogenic analysis

Molecular assay

Cytochemical stains (differentiate

abnormal cells)
Flow cytometry
 Hematological Diseases (Diseases may be classified in more
than one category)
Diseases of improper or insufficient production
Anemias Examples include iron deficiency anemia
and aplastic anemia
Leukemias WBC production is affected and
defective. Examples include acute and chronic
leukemias
Thrombocytopenia Decreased platelets. Patient
may have bleeding problems
 Diseases of defective cell function
May be combination of improper cell production and
defective function
Examples include iron deficiency anemia and certain
leukemias
Inherited hematological diseases
Examples include hemophilia, sickle cell anemia, G6PD
deficiency, thalassemias
Secondary or acquired hematological diseases
Examples include hemolytic disease due to renal
pathologies or atypical lymphocytes due to viral infections