SECTION TWO 



violet to purple granules. Malarial parasites and Leishmania: 

 chromatin, red ; cytoplasm, blue. Trypanosomes : chromatin, red. 



Note: The timing of the staining either before or after dilution 

 may be altered to suit individual requirements. Staining effects 

 similar to Giemsa are obtained by staining for ten minutes in 

 Leishman stain diluted with twice its volume of distilled water 

 buffered to pH 6-5. 



Reference: Leishman, W. B. (1901). 



LEUCO PATENT BLUE 

 For the identification of haemoglobin. 



Solutions required: 



A. Patent blue AF54 . . . . i gm. 

 Distilled water . . . . . . 100 ml. 



Dissolve; then add: 



Zinc metal powder . . . . 10 gm. 



Glacial acetic acid . . . . 2 ml. 



Boil until the blue colour completely disappears. 

 Allow to cool ; shake with about i gm. of decolorizing 

 carbon; then filter. The liquid which should then 

 be quite colourless is stored in a well stoppered 

 bottle. 



B. Solution A . . . . . . 10 ml. 



Glacial acetic acid . . . . 2 ml. 



Hydrogen peroxide 3% . . i ml. 



N.B. : This solution must he freshly prepared and 

 filtered before use. 



C. Safranin o*i% aqueous . . 99 ml. 

 Glacial acetic acid . . . . i ml. 



Technique: 



1. Fix tissue blocks, not more than 3 to 5 mm. in thickness in 

 10% formalin buffered to pH 7-0 for 24 to 28 hours (prolonged 

 fixation should be avoided). 



2. Embed in paraffin wax as usual and cut section 5 to 6/x in 

 thickness. 



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