Blood is an important specimen in the clinical

laboratory. The importance of blood in medical

branches:
Microbiology: to detect systemic infection of
bacteria through immunoassay in blood culture.
Liver and other organ: specific enzyme in each
site of organ can be warning for damaged
tissue/organ (not normally found in blood/low in
blood)
Hematology: abnormal cell count and
concentration, morphology can indicate blood
disease and disorder (anemia, leukemia,
thrombopenia, malignancy)
Indication of certain disease: disorder,
deficiency, or toxicity.
e.g.: Blood glucose increase = diabetes mellitus
Cholesterol, LDL, triglyceride increase = obese,
cardiac failure
Blood protein decrease = malnutrition,
kwashiorkor
- Pharmacology: response of drug
- Radiology: response of radiation

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Basic Examination of Blood

Advantages: Easy to collect, carries important info

about patients condition, many tests can be carried
out

COMPLETE BLOOD COUNT (CBC)

Provides important information about the kinds
and numbers of RBC, WBC, platelet
Parts of routine physical examination
Help to: Evaluate symptoms (weakness, bruising,
fever, weight loss)
Diagnose condition (anemia, infection)
Diagnose diseases of the blood (leukemia)
Monitor response to some types of
drug/radiation treatment
Results: Normal value can vary from lab to lab.
Normal value for CBC tests vary, depending on age,
sex, elevation above sea level, and type of sample.

1.
Hemoglobin
The main component of RBC, gives RBC its red color.
Conjugated protein; contain 2 pairs of polypeptide
(globin) and 4 prosthetic heme group.

Examination aim: measure the amount of

hemoglobin in blood
Clinical implication: E.g. Hb low anemia
Determining concentration of Hb with
Hemiglobincyanide (HbCN) method *hazard to
environment

Principle:
Blood is diluted in a solution of potassium
ferricyanide (K3Fe(CN)6) then measured by
spectrophotometer at 540 nm and compared
with that of a standard Hi CN solution.
Errors in hemoglobinometry : error inherent in the
sample, method, equipment, operators error

2.
Hematocrit
Explanation: ratio of volume of erythrocytes to whole
blood. Expressed in % (conventional) or decimal
fraction L/L. (SI units)

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Main function: transport O2 from lungs (conjugated

protein: vehicle for transport of O2 & CO2).
Derivatives of hemoglobin: hemiglobin
(methemoglobin), sulfhemoglobin, carboxyhemoglobin
(most)

Venous puncture shows same result as skin puncture

both greater than body hematocrite. It reflects
concentration of red cells not total cell mass
Clinical implication: green/orange plasma:
hemoglobinemia
(rising number of Hb)
low Ht = anemia, high Ht=
polycythemia
Method & units: mix thoroughly before taking
sample
Direct: centrifugation with macro/micromethod
Indirect: automated instrument MCV x RBC
count
Sources of error: centrifugation, sample, etc.
3.
Blood cell counting
Expressed as concentration cells per unit volume
of blood (mm3/L)
CBC can be performed:
Manually :
Hemocytometer
Calculate from other CBC results (RBC
indices)
Semiautomatically
Automatically (hematologic analyzer)
a) WBC count

To calculate number of components of WBC; help

to identify infection (neutrophilia, lymphocytosis)
and allergic or toxic reaction to certain medication
(eosinophilia)
Expressed as concentration cells per unit volume
of blood (mm3)
Components: PMN, band neutrophil, lymphocyte,
monocyte, eosinophil, basophil
Principle: anticoagulant EDTA + diluting fluid to
lyse.
Clinical aspect: high WBC=leukocytosis, low WBC
= leucopenia.

b) WBC differential counts

Measure the percentage of each type of white
blood cells
Components: PMN, band neutrophil (include
immature neutrophil), lymphocyte, monocyte,
eosinophil, basophil
Expressed as a percentage of each type
Increase/decrease number of each type help to
identify : Infection (neutrophilia, lymphocytosis);
Allergic or toxic reaction to certain medication
(eosinophil/basophil); Malignancy (leukemia)

c) RBC cell count

Count number of RBC in every unit of blood
volume

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Expressed as concentration cells per unit volume

of blood (mm3).
Clinical implication: < normal anemia, higher
polycythaemia
Methods & units: manual hemocytometer
chamber, can use semiautomated method

d) RBC indices
Determine the size of RBC, Hb content and
concentration in the RBC
Useful for morphologic characterization of anemia
Maybe calculated from: Red cell count, Hb
concentration, or Hematocrit
3 RBC indices :
MCV (Mean Corpuscular Volume)
Shows the size of RBCs, average volume of RBC
Classified as: normocyte, microcyte, macrocyte
Calculated from the Ht and RBC count
Expressed in femtoliter (fL) or cubic micrometers
Diurnal variation: highest in the morning, lowest
in the evening

MCV = Ht 1000 / RBC (in million

per L)

MCH (Mean Corpuscular Hemoglobin)

Is the content (weight) of Hb of the average
RBCs.

Calculated from the Hb concentration and RBC

count.
Expressed in picograms (g) or pg
Clinical implication: MCH low=Hypochromic RBC,
MCH high= Hyperchromic RBC (reticulocyte)

MCHC (Mean
Hemoglobin
MCH Corpuscular
= Hb (g/L) /RBC
(in million
Concentration)
per L)
Is the average concentration of Hb in a given
volume of packed red cells
Calculated from Hb concentration and the
hematocrit.
Expressed in g/dL
Clinical implication: MCHC low: Hypochromic
RBC
MCHC = Hb (g/dL)
/Hct

e) Platelet count
Platelets normally are 2-4 m in diameter, 5-7 fL in
volume
Function: Homeostasis, maintain vascular integrity,
blood clotting, more difficult to count (small, tendency
to adhere to glass and one another), MPV (Mean
Platelet Volume) can also be counted, women have
greater number of platelet than men

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Is expressed as concentration cells per unit

volume of blood (mm3)
Clinical implication: platelet increase:
thrombocytosis; decrease: thrombocytopenia

** Reticulocyte count
Characteristic: immature non-nucleated red cells,
contain RNA, continue to synthesze Hb
after loss of nucleus
Clinical implication: increase reticulocyte=increase
production of RBC

4.
Blood film examination
Principle: blood spread in glass slide/covering
Aim: to evaluate the numbers, size, and shape of
RBC, WBC and platelet in a stained smear of
peripheral blood
Red Cell Distribution Width (RDW)

Reports whether all the red cells are about the

same width, size, and shape.

This helps further classify the types of anemia.

5.
Erythrocyte Sedimentation Rate (ESR)
Useful, non-specific marker of inflammation
Measures:

The settling of erythrocytes in diluted human

plasma over a specified time period (1 hour)
- Distance from the bottom of the surface
meniscus to the top of erythrocyte
sedimentation in a vertical column containing
diluted whole blood.
Normal values :
Adult men
0-15 mm/h
Adult women
0-20 mm/h
Methods:
- Manual :
o Westergren Method
o Wintrobe & Landsberg Method
- Automatic

Factors affecting:
- RBC size & shape
Rouleaux increase ESR
Rate: microcyte macrocyte
Spherocyte decrease ESR
Sickle cell decrease ESR
- Plasma fibrinogen & globulin levels
Presence of fibrinogen increase ESR
Fibrinogen decrease (-) charge (zeta potential) of
erythrocytes that keep them apart. If zeta
potential decrease, rouleaux will be formed.
Excess immunoglobulin increase ESR.
- Mechanical
- Technical

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Conditions associated with an elevated ESR

Rheumatoid arthritis, Multiple Myeloma,
Cryoglobulinemia, Temporal arteritis, inflammatory
diseases, Pregnancy, Anemia, Malignant neoplasms,
Paraproteinemias, Macroglobulinemia,
Hyperfibrinogenemia, Chronic infections, Collagen
disease, Polymyalgia rheumatica

OTHER EXAMINATIONS USING BLOOD

SPECIMEN
Liver function tests are one of the blood tests
that are most commonly performed to assess the
function of the liver or injury caused to the liver. Liver
damage is detected initially by performing a simple
blood test that determines the level of various liver
enzymes present in the blood. The most widely used
liver enzymes that are sensitive to abnormalities in
liver and are most commonly measured are the

SGOT and SGPT are highly sensitive markers of

liver damage due to various diseases or injury.
However, the fact is that higher than normal levels
should not be automatically considered as indicative
of liver damage. They may or may not imply liver
disease. For instance, these enzymes are also
elevated in cases of muscle damage.
SGOT
Normally present in a number of tissues such as
heart, liver, muscle, brain and kidney. It is released
into the blood stream whenever any of these tissues
gets damaged.

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aminotransferases. The two aminotransferases that

are checked are the alanine aminotransferase (ALT
or SGPT) serum glutamic pyruvic transaminase
and aspartate aminotransferase (AST or SGOT)
serum glutamic oxaloacetic transaminase.

Blood AST level is increased in conditions of

muscle injury and heart attacks. Hence, it is not highly
specific liver tissue damage indicator as it can be
elevated in conditions other than liver damage.
The normal levels of SGOT is in between 5 and 40
units per liter of serum.
SGPT
Normally present in large concentrations in the
liver. Hence, due to liver damage its level in the blood
rises, thereby, serving as a specific indicator for liver
injury.
The normal levels of SGPT in between 7 and 56
units per liter of serum.

However the normal ranges of SGPT and SGOT

differ depending on the protocols and technique used
to measure in laboratory.