BLOOD

Functions of Blood:
• Supply of Oxygen to tissues and removal of
CO2
• Supply of vital nutrients such as glucose,
amino acids, and fatty a dissolved in blood or
bound to plasma proteins.
• WBC’s Present in the blood responsible for
immunological functions.
• Coagulation factors –homeostatis and blood
clotting
 Functions of Blood:

• Messenger molecules such as hormones

secreted by endocrine system are transported
to the target organ via blood.
Composition of blood
Haematopoiesis
 HEMATOLOGICAL TESTS
• Common Hematology Laboratory Tests
– Complete Blood Count (CBC) includes:
• RBC count
• WBC count
• Hemoglobin
• Hematocrit
• WBC Differential
• Platelet count
• Additional information on size and appearance
of blood cells
• Common Hematology Laboratory Tests
– Erythrocyte Sedimentation Rate (ESR)
– Reticulocyte Count
– Sickle Cell Testing
– Coagulation Tests include:
• Prothrombin Time (PT)
• Partial Thromboplastin Time (PTT)
• Fibrinogen
• Factor Analysis
 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, among others
• 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
Hemoglobin is a large complex protein. It has a molecular weight
of 64,000 Daltons, Roughly spherical in shape.

The two pairs of

globin chains are
twisted together so
that the large heme
groups are exposed
to the exterior of the
molecule.

Each globin chain contains 141 amino acids. Each hemoglobin

molecule consists of four heme groups bound to each of four
globin chains & may carry up to four molecules of Oxygen.
 Functions of Hemoglobin
• Oxygen delivery to the tissues
• Reaction of Hb & oxygen
 Oxygenation not oxidation
 One Hb can bind to four O2 molecules
 Less than .01 sec required for oxygenation
 β chain move closer when oxygenated
 When oxygenated 2,3-DPG is pushed out
 β chains are pulled apart when O2 is unloaded,
permitting entry of 2,3-DPG resulting in lower
affinity of O2
Oxy & Deoxy-hemoglobin
Transport of CO2 :The Bohr Effect
Disorders of Hemoglobin:

1. Amino acid substitutions

2. Amino acid deletion
3. Elongated globin chain resulting from
chain termination, mutation
4. Fused or hybrid chains
 DETERMINATION OF HEMOGLOBIN
1.Cyanmethemoglobin (Potassium Ferricyanide)

2. Sodium Lauryl Sulphate (Cyanide free)

CYANIDE POISONING - Cyanide inhibits the last

site of the electron transport chain . It prevents the
passage of electrons by binding to a specific
component of the chain (cytochrome a3) blocking the
redox reaction. Thus all electron carriers before the
block will be fully reduced whereas those after the
block will be oxidized.
 CORAL Hb KIT
1) Cyanmethemoglobin (potassium Ferricyanide)
A) Hemocor –C
B) Hemocor –D

2) Hemosafe SLS
 Principle
• Potassium ferricyanide converts hemoglobin
in the sample to methemoglobin.
• Methemoglobin further reacts with potassium
ferricyanide to form cyanmethemoglobin
complex.
• Colour produced directly propotional to the
hemoglobin present.
 Cyanmethemoglobin
(potassium Ferricyanide)
• Hemocor –D ; ready to use;
• Hemocor C –W.R to be prepared from conc
reagent (1:25)
• Room Temperature stable
• Reaction time – 3 minutes
• End color stable for 24 hours
• High linearity – 20gm/dl
• Result via factor / standard
• Hb reading chart for standard available
 Hemosafe -SLS
Principle
• SLS disrupts the RBC membrane to yield a
SLS-hemoglobin complex.
• This complex id read photometrically at
540nm.
 Benefits
• Hemosafe reagent- ready to use;
• Room Temperature stable
• Reaction time – 5 minutes
• End color stable for 24 hours
• High linearity – 20gm/dl
• No need of running a standard
• Hemoglobinis is less affected by presence of
leukocytes with Hemosafe-SLS ,ethod as
compared to cynmethemoglobin method