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  Section: Biotechnology Methods » Cell Biology and Genetics
 
 
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Differential Staining of Blood

 
     
 
Content
Cell Biology and Genetics
  Cell Cycles
  Meiosis in Flower Buds of Allium Cepa-Acetocarmine Stain
  Meiosis in Grasshopper Testis (Poecilocerus Pictus)
  Mitosis in Onion Root Tip (Allium Cepa)
  Differential Staining of Blood
  Buccal Epithelial Smear and Barr Body
  Vital Staining of DNA and RNA in Paramecium
  Induction of Polyploidy
  Mounting of Genitalia in Drosophila Melanogaster
  Mounting of Genitalia in the Silk Moth Bombyx Mori
  Mounting of the Sex Comb in Drosophila Melanogaster
  Mounting of the Mouth Parts of the Mosquito
  Normal Human Karyotyping
  Karyotyping
  Black and White Film Development and Printing for Karyotype Analysis
  Study of Drumsticks in the Neutrophils of Females
  Study of the Malaria Parasite
  Vital Staining of DNA and RNA in Paramecium
  Sex-Linked Inheritance in Drosophila Melanogaster
  Preparation of Somatic Chromosomes from Rat Bone Marrow
  Chromosomal Aberrations
  Study of Phenocopy
  Study of Mendelian Traits
  Estimation of Number of Erythrocytes [RBC] in Human Blood
  Estimation of Number of Leucocytes (WBC) in Human Blood
  Culturing Techniques and Handling of Flies
  Life Cycle of the Mosquito (Culex Pipiens)
  Life Cycle of the Silkworm (Bombyx Mori)
  Vital Staining of Earthworm Ovary
  Culturing and Observation of Paramecium
  Culturing and Staining of E.coli (Gram’s Staining)
  Breeding Experiments in Drosophila Melanogaster
  Preparation of Salivary Gland Chromosomes
  Observation of Mutants in Drosophila Melanogaster
  ABO Blood Grouping and Rh Factor in Humans
  Determination of Blood Group and Rh Factor
  Demonstration of the Law of Independent Assortment
  Demonstration of Law of Segregation

To identify different stages of white blood cells in human blood.

Introduction
White blood corpuscles (WBCs) or leucocytes, are colorless, actively motile, nucleated living cells. They are variable in size and shape and exhibit characteristic amoeboid movement. Under certain physiological and pathological conditions, WBCs can come out of the blood vessels by a process called diapedesis. The lifespan of WBCs is 12–15 days. Unlike red blood corpuscles, leucocytes or WBCs have nuclei and do not contain hemoglobin. They are broadly classified into 2 groups:
  • Granulocytes
    and
  • Agranulocytes
Granulocytes develop from red bone marrow and agranulocytes from lymphoid and myeloid tissues. Granulocytes are subdivided into 3 groups:
  1. Eosinophils
  2. Basophils
    and
  3. Neutrophils
Agranulocytes are subdivided into 2 groups:
  1. Lymphocytes
    and
  2. Monocytes
The main features of WBCs are:
  1. Phagocytosis
  2. Antibody formation
    and
  3. Formation and secretion of lysozyme, heparin, etc.
Principle
WBCs are the type of blood cells that have a nucleus but no pigment. They are important in defending the body against diseases because they produce antibodies against any foreign particle or antigens. WBC can be divided into 2 types—granulocytes and agranulocytes. Granulocytes consists of neutrophils, eosinophils, and basophils.
Neutrophils. Do not stain with either acidic or basic dyes. They have manylobed nucleus and are called polymorphonuclear leucocytes or polymorphs. They constitute about 76.9% of the total leucocytes found.

Eosinophils. A lobed nucleus is present, with cytoplasmic granules that stain with acidic dyes. They constitute 1%–4% of the total leucocyte count. Basophils. It has a lobed nucleus and the cytoplasm contains granules, which stain with basic dyes. It comprises 0%–4% of the total leucocyte count. Agranulocytes. Consist of lymphocytes and monocytes. Lymphocytes. This is a type of WBC with a very large nucleus. It is rich in DNA and a small amount of clear cytoplasm is present.


Procedure
  1. The fingertip is cleaned with cotton dipped in alcohol and pricked with a sterilized needle.
  2. One or two drops of blood are placed on the right side of the slide, and with the help of another clean slide, the blood is drawn so that a thin smear is formed and dried.
  3. The dried smear is fixed using zinc acetone, 3 methyl alcohol, or absolute alcohol for 5 min.
  4. The smear is dried and stained with Giemsa or Leishman stain.
  5. Subsequently, the distilled water (double the amount of water of the stain) is added onto the same smear slide.
  6. The smear is mixed using a pipette for 10–15 min.
  7. The slides are kept in running water to remove excess stain, and then dried.
  8. The slide is observed under an oil immersion lens.

 
     
 
 
     




     
 
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