GIEMSA STAIN
INTRODUCTION
Giemsa stain is a group of Romanowsky stain. It is a gold standard staining technique used for
both thin and thick smears to examine blood for malarial parasites and to morphologically
differentiate the nuclear and cytoplasm Erythrocytes, leucocytes and platelets and parasites. It
is also used in cytogenetics and histopathology for the diagnosis of:

Malaria, spirochetes and other blood parasites

Chlamydia trachomatis inclusion bodies

Borrelia species

Yersinia pestis

Histoplasma species

Pneumocyctis jiroveci cycts
OBJECTIVES

To permit differentiation of cells present in hematopoitic tissue.

To demonstrate of some microorganisms.
PRINCIPLE
The neutral dyes combining the basic dye methylene blue and the acid dye Eosin, give a wide
color range when staining. The pH of the staining solution is critical and ideally should be
adjusted for different fixatives. More acid pH levels give more selective chromatin staining
and less cytoplasmic basophilia; less acid pH levels gives denser nuclei and increased
cytoplasmic basophilia.
REAGENTS REQUIRED
1. Methanol
2. Giemsa powder
3. Glycerine
4. Water (buffer)
PREPARATION OF THE GIEMSA STAIN STOCK SOLUTION (500ML)
1. Into 250ml of methanol, add 3.8g of Giemsa powder and dissolve.
2. Add 250ml of glycerine to the solution, slowly.
3. Place the bottle of stain in water at 50-60°C or at 37°C for upto 2 hours with frequent
mixing.
4. Label the bottle and store in a cool, dark place with a firm stopper.
PREPARATION OF WORKING SOLUTION

Add 10ml of stock to 80ml of distilled water and 10ml of methanol.
STAINING PROCEDURE
1. Prepare blood film on a grease free slide and air dry.
2. Treat the dried blood film with methanol for 3-5 minutes.
3. Immerse the slide in the staining fluid containing 30 drops (0.67ml) of giemsa stain in
30ml of distilled water and stain it for 30-40 minutes.
4. Wash with distilled water allowing the preparation to differentiate for 1-3 minutes.
5. Air dry the film and examine under microscope.
Note: According to the Himedia stain
RESULTS
Nuclei
Blue
Cytoplasm of leukocytes
May be shades of pink, grey, or blue, depending on cell
type and development
Bacteria
Blue