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)

rainwater@mymelody.com || 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)

rainwater@mymelody.com || 1st semester, AY 2011-2012

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%

rainwater@mymelody.com || 1st semester, AY 2011-2012