SOP – Malaria Microscopy - NVBDCP

ConstituentsQuality Assurance of Malaria Diagnostic testsThe J.S.B. stain comprises of two solutions, J.S.B.I and J.S.B.II. The composition ofeach staining solution is as follows:J.S.B. Solution I• methylene blue (Medicinal) 0.5 gm• sulphuric acid (H 2 SO 4 ) 1% 3.0 ml• potassium dichromate (K 2 Cr 2 O 7 ) 0.5 gm• disodium hydrogen phosphate 3.5 gmdihydrate (Na 2 H PO 4 2 H 2 O)• distilled water 500 ccJ.S.B. solution II• eosine 1.0 gm• distilled water 500 ccBuffered water8.3.3.4 Others• disodium hydrogen phosphate dihydrate 0.22 gm(Na 2 H PO 4 2 H 2 O)• potassium acid phosphate (KH 2 PO 4 ) 0.74 gm• distilled water 1000 ml• enamel tray ( for keeping the staining kits)• heating mantle/stove for boiling stain• filter paper for filtering the stains for day to day use• soft cloth8.3.4 Method of preparation of JSB stain8.3.4.1 JSB I• take 500 ml of distilled water in a flat bottom round flask (1000 ml), add 0.5gm methylene blue and stir the whole solution in order to get it dissolved.• further, add 3 ml of 1% sulphuric acid in three equal parts with stirring toensure thorough mixing.• add potassium dichormate (0.5 gm), which forms a purple precipitate ofmethylene blue chromate by oxidation of dye in acid medium.• subsequently, add 3.5 gm of disodium hydrogen phosphate dihydrate andagain stir the whole solution thoroughly, ensuring that the precipitatedissolves.• then boil the solution with reflux condenser for one hour.• keep the solution thus prepared overnight at room temperature for 24 hoursfor maturation before use.SOP – Malaria Microscopy© Copyright to Dte. NVBDCP Only. Any modification are prohibited