ROMANOWSKY STAIN-types,uses, preparation

Romansky stain definition,composition,uses,types,preparation,result

1. ROMANOWSKY STAIN Dharshini.M Phd Scholar pathology

2. Contents Definition Composition Principle Result Types Stain preparation and its uses

3. Definition Universally used routine stains for blood films. Neutral stain It contains both positively & negatively charged particles which imparts different colors to different components . It stains red cells, white cells, platelets, parasites and bone marrow.

4. Composition Basic dye –Methylene blue Acidic dye-Eosin Y

5. Principle It works on the principle that “the acidic components of the cells are stained by the basic dye methylene blue (azure) forming a blue-purple colour while the basic components of the cell are stained by the acidic dye (eosin Y) forming orange-red colouration ”

6. Result ACIDIC COMPONENTS Nucleus (DNA) Some cytoplasmic components (RNA, Granules ) Imparts-blue BASIC COMPONENTS Cytoplasm Mitochondria Secretory granules Imparts –orange red

7. Types Leishman stain May Grunwald Stain Giemsa stain Wright stain Jenner stain Field stain

8. Leishman stain It is a polychromatic stain Differential count Components Methanol : Fixes cell Methylene blue : stains RNA , DNA (blue) Eosin : stains Hb , Eosin granules

9. Preparation 0.15 g of powdered dye + 100 ml of methanol Place 50`C in water bath for 15 minutes . Cool and filter

10. Procedure Staining solution 10 to 15 drops on the smear. Wait for 1 minute. Add gently equal number of drops of buffered water using a dropper. Wait for 10 minutes. Wash the slide with tap water and wait for dry Examine under 10x, 40x and 100x

11. May Grunwald Stain Polychromatic stain Used routinely for staining of air-dried cytological smears, blood, and bone marrow smears. It is useful for studying cellular morphology and is superior to PAP stain for studying cytoplasm, granules, vacuoles, and basement membrane .

12. Preparation MG stain preparation 0.3 g of powdered dye + 100 ml of methanol Place 50`C in water bath for 15 minutes. Cool at 20`C for 24 hours and filter. Giemsa stain preparation 1 g of powdered dye + 100 ml of methanol Place 50`C in water bath for 15 minutes. Cool and filter

13. Procedure Prepare a thin smear and air dry. Fix smears for 5-10 minutes with methanol. Stain the smear in May Grunwald working solution for 10 minutes. Rinse in pH 6.8 buffer. Stain the slides with diluted Giemsa stain for 30 minutes. Wash the smears with distilled water and let them dry. Mount the slide with DPX and examine under microscope.

14. GIEMSA STAIN It is a stain used in cytogenetics and parasite examination. This consist of number of compound made by mixing different proportion of methylene blue and eosin. Preparation 1 g of powdered dye + 100 ml of methanol Place 50`C in water bath for 15 minutes. Cool and filter

15. Wright stain Used to differentiate nuclear/ cytoplasmic morphology of platelets , RBC, WBC and parasites . It became widely used for performing differential white blood cell counts, which are routinely ordered when conditions such as infection or leukemia are suspected. PREPARATION Wright stain powder -0.25gm Acetone free methanol-100ml Dissolve powder in methanol.Keep for few days { for ripening}before using.