02 - Examination of Blood and Bone Marrow Hematology

2 Examination of Blood and Bone Marrow Hematology
Hematology
Study of blood cells and coagulation
Encompasses analysis of blood cells
Concentration structure and function of

cells in the peripheral blood
Precursors in the bone marrow
Chemical constituent of plasma or serum
Bone Marrow Examination
Morphology of the cellular elements

(peripheral smear)
Cellularity of the bone marrow

Distribution of cells
Maturation of cells
Presence of rare cell types or abnormal cells
General Characteristics of Blood
Interpretation
Cellularity of bone marrow

Numbers of megakaryocytes (platelet precursor)
Myeloid:Erythroid ratio
Cytologic or maturation abnormalities

Storage of iron and proportion of sideroblast
Quantity
Varies with each individual, but the mean average in

the adult approximates 8% of the body weight (or
around 1/13 of the total body weight) or 5 to 6 liters
of blood or roughly estimated at 75cc/kg body weight
Color
Is due to the presence of hemoglobin in the RBCs
Arterial blood
Venous
Bright red
Purplish red
Coal gas poisoning
Bright cherry red
Potassium chlorate
poisoning
Chocolate
Oxyhemoglobin
Reduced
hemoglobin
Carbon monoxide
-hemoglobin
Methemoglobin
Reaction
Blood is slightly alkaline, having a pH of 7.4 to 7.45.

This alkalinity remains constant throughout life.
Specific Gravity
Ranges from 1.075. It slightly varies with sex, being
higher in males than females. In pathological

condition, this is greatly altered.
Viscosity
Viscosity of blood approximates 5-6 times than of H2O
Tests used in hematological analysis

Blood
CBC Platelet count

Blood indices
Reticulocyte count
Bone Marrow Examination (Aspiration and Biopsy)
Marrow films
Direct films
Imprints
Crust preparations
Special staining methods
Romanowskys stain
Perls test for iron

Blood Analysis
CBC examination
Hgb
Hct
RBC count
WBC differential count

Platelet count infection
Blood indices
Peripheral smear
Reticulocyte count index of erythropoiesis
Indications
Microcytic anemia (iron deficiency anemia)
Macrocytic anemia (megaloblastic anemia) VIt.

B12
Elevated reticulocytes (hemolytic anemia)
Neutropenia, thrombocytopenia and pancytopenia

aplastic anemia
Immunoglobulin abnormalities such as plasma cell
myeloma or macroglobulinemia
Hemoglobin
Main component of the red blood cell

A conjugated protein composed of 2 pairs of
polypeptide chains (globins) and 4 prosthetic heme

groups
Each gram of Hgb holds 1.34 ml of oxygen

The red cell mass of the adult contains approximately
600 grams of hemoglobin capable of carrying 800 ml

of oxygen
Complete Blood Count (CBC)
Sahlis Method (Hemoglobin Determination)
Principle: Conversion of Hgb to acid hematin by the

addition of 0.1 N Hcl
Normal Range
Conventional = 12.0 16.0 g/dL
SI = 120-160 g/L
Indications
Anemia
Polycythemia
Cyanmethemoglobin (Hemoglobin Determination)
Principle: Ferricyanide oxidizes oxyhemoglobin and

cyanide converts methemoglobin to
cyanmethemoglobin
Measures all types of Hgb except sulf-Hgb
Normal range:
Convention: 120 160 g/dL
SI: 120 160 g/L

Hemoglobin Functions
Transport of oxygen from the lungs (high oxygen
tension) to the tissues (low oxygen tension)
[email protected] || 1st semester, AY 2011-2012
Types
Reduced Hgb
Hgb with iron unassociated with oxygen
Oxyhemoglobin
Hgb with iron associated with one molecule

of O
Methemoglobin
Hgb with increased H1 in the erythrocytes

Sulf-hemoglobin
Mixture of oxidized partially denatured
forms of hgb that form during oxidative

hemolytic process
Oxidation of Hgb sulfur incorporation
Heme ring (Hgb)
green
hemochrome oxidation
denaturation and
precipitation of Hgb
(Heinz bodies)
Implications
Sulfhemoglobin cannot transport oxygen

but it can combine with carbon monoxide
to form carboxysulfhemoglobin
Methemoglobin and sulfhemoglobin cannot be
reduced back to hemoglobin and it will remain in the
cells until the cells breakdown
Erythrocyte Count
Principle : Blood is diluted with fluid that is isotonic

with erythrocytes
Formula
RBC count =
# of cells
counted_____
Area x depth x dilution
factor
N.V.
Conventional: 4,500,000 6,000,000/cumm

12
SI: 4.5 X 6.0 X 10 /L

Leucocyte Count
PRINCIPLE: Blood is diluted with a fluid that lyzes the

non-nucleated RBC but not the nucleated RBC
FORMULA
WBC count =
# of cells
counted_____
Area x depth x dilution
factor
N.V.
Conventional: 4,500 10,000/cumm

9
SI: 4.5 X 10.0 X 10 /L

RBC Morphology
Assessment of Red Blood Cell Morphology
1. Peripheral smear
2. Blood Indices
RBC MORPHOLOGY: Peripheral Smear
1. Size
2. Shape
3. Hemoglobin Content
4. Inclusions
5. Immaturity
Normal:
Size
Shape
6-8 um
Biconcave disc; flexible and
deformable
Central pallor - small
Hemoglobin Content
* Azurophilic granules
* Absent nuclei in erythrocytes and platelets
* Normal erythrocyte/platelet ratio 15-20:1 (helpful index for
recognition of thrombocytopenia and thrombocytosis)
Blood Indices
1. Size
2. Hemoglobin content
Changes in shape are more difficult to assess expertise
needed to have good knowledge of what constitutes normality
Allows classification of RBC as to size, hemoglobin content

and concentration
1. MCV (Mean Corpuscular volume)
2. MCH (Mean Corpuscular Hgb)
3. MCHC (Mean Corpuscular Hgb Concentration)
Anemia
Reduction in the concentration of Hgb or Hct level in

the peripheral blood below the normal (lower limit of
95% of reference interval for individual age, sex and
geographical location [altitude])
Causes:
1. Impaired red cell production
2. Blood loss
3. Accelerated red cell destruction (hemolysis)
in excess of the ability of bone marrow to
replace these losses
Classification based on:

Mean corpuscular volume (MCV)
Is the average volume of individual RBC
Allows classification of cells into normocytic,
macrocytic and microcytic

MCV = __ HCT x_10__
12
RBCs (x 10 /L)
NV = 87 5 cu microns (fl)
(82 92 fL)
Mean corpuscular hemoglobin (MCH)
Is the amount of Hgb by weight in an average RBC
Allows classification of cells into normochromic,

hypochromic and hyperchromic
In newborn and macrocytic anemia
- MCH is high
In deficiency anemia,
- MCH is low
MCH = __Hgb gm% x 10__
12
RBCs (x 10 /L)
NV = 27-31 ug (pg)
Mean corpuscular hemoglobin concentration (MCHC)
Is the concentration of hemoglobin in an average RBC

If below normal, hypochromic
Higher than normal concentration are not possible as
normal RBC contains the maximum amount of Hgb

Normochromic, within normal MCHC
MCHC = __Hgb gm % x 100__

Hct
NV = 33-38%
No such thing as hyperchromacity (cannot be

secondary to high MCHC) for RBC membrane
abnormalities
Mean Corpuscular Hemoglobin (MCH) and

Mean Corpuscular Hemoglobin Concentration (MCHC)
Normochromic
MCH
27 31
pg
MCHC
33
38%
Hypochromic
< 27 pg
< 33%
Hyperchromic
> 31 pg
> 38%

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2 Examination of Blood and Bone Marrow Hematology

Study of blood cells and coagulation

Encompasses analysis of blood cells

Concentration structure and function of

Precursors in the bone marrow

Chemical constituent of plasma or serum

Bone Marrow Examination

Morphology of the cellular elements

Cellularity of the bone marrow

Presence of rare cell types or abnormal cells

General Characteristics of Blood

Cellularity of bone marrow

Cytologic or maturation abnormalities

Varies with each individual, but the mean average in

Is due to the presence of hemoglobin in the RBCs

Coal gas poisoning

Bright cherry red

Blood is slightly alkaline, having a pH of 7.4 to 7.45.

higher in males than females. In pathological

Viscosity of blood approximates 5-6 times than of H2O

Tests used in hematological analysis

CBC Platelet count

Bone Marrow Examination (Aspiration and Biopsy)

Perls test for iron

WBC differential count

Reticulocyte count index of erythropoiesis

Macrocytic anemia (megaloblastic anemia) VIt.

Neutropenia, thrombocytopenia and pancytopenia

Main component of the red blood cell

polypeptide chains (globins) and 4 prosthetic heme

Each gram of Hgb holds 1.34 ml of oxygen

600 grams of hemoglobin capable of carrying 800 ml

Principle: Conversion of Hgb to acid hematin by the

Conventional = 12.0 16.0 g/dL

Cyanmethemoglobin (Hemoglobin Determination)

Principle: Ferricyanide oxidizes oxyhemoglobin and

Convention: 120 160 g/dL

SI: 120 160 g/L

tension) to the tissues (low oxygen tension)

[email protected] || 1st semester, AY 2011-2012

Hgb with iron associated with one molecule

Hgb with increased H1 in the erythrocytes

forms of hgb that form during oxidative

Sulfhemoglobin cannot transport oxygen

Principle : Blood is diluted with fluid that is isotonic

Conventional: 4,500,000 6,000,000/cumm

SI: 4.5 X 6.0 X 10 /L

PRINCIPLE: Blood is diluted with a fluid that lyzes the

Conventional: 4,500 10,000/cumm

SI: 4.5 X 10.0 X 10 /L

Allows classification of RBC as to size, hemoglobin content

Reduction in the concentration of Hgb or Hct level in

Classification based on:

Allows classification of cells into normocytic,

macrocytic and microcytic

Is the amount of Hgb by weight in an average RBC

Allows classification of cells into normochromic,

[email protected] || 1st semester, AY 2011-2012

Mean corpuscular hemoglobin concentration (MCHC)

MCHC = Hgb gm % x 100