====WDI OF PREPARATION OF STAINING====

===GIEMSA STAIN===

STOCK SOLUTION\\
Buffer pH 6.8 – 
                KH2PO4 (9.1 G) + D.W. (1016 ml)
			+
		Na2HPO4.2H2O (11.5 g) + D.W.( 984 ml)
Buffer pH 7.0 - 
                KH2PO4 (3.5 G) + D.W. (389 ml)
			+
		NaH2PO4.2H2O (7.1 g) + D.W.( 611 ml)
Buffer pH 7.2 - 
                KH2PO4 (2.5 G) + D.W. (280 ml)
			+
		NaH2PO4.2H2O (8.4 g) + D.W.( 720 ml)
WORKING SOLLUTION:\\
For Working solution of buffer ( 25ml of stock solution + 475 ml D.W.)\\
Buffer pH 6.8 – For Bone marrow staining\\
Buffer pH 7.0 – For P.S. staining\\
Buffer pH 7.2 – For Malarial parasite staining\\

Dilution:\\
For B.M. Staining - 1:6.5 ( 10 ml Geimsa Stain + 55 ml Buffer water)\\
For P.S. Staining – 1:5 ( 10 ml Geimsa Stain + 40 ml Buffer water)\\

PREPARATION OF STAIN:\\
• Take 5 gm of gimsa powder and crush it properly, then mix it in 500 ml of glycerol and keep it for 48 hours.\\
• Add & well mix 500 ml of absolute methanol in the above mentioned solution and keep at room temperature for one week.\\
• After filtering it is stored in amber colored bottle at room temperature\\

__STAINING PROCEDURE:__\\
• Add buffer( 6.8- 7.2 pH) in the filtered stain in 1:4 dilution\\
• Arrange thick/ thin smears on double rod stand.\\
• Pour the stain-buffer mixture on the arranged slides. Leave it for 10-15 minutes\\
• Stain over the slides should be mixed by blowing with pipette.\\
• After 10-15 minutes, wash the slides in slowly running tap water.\\
• Dry the washed slides in air at room temperature.\\