Histological Staining Techniques

Leishman's stain for blood cells and parasites

Leishman 1901.

Prepare cytospin slide or smear of sample.

Red blood cells - red to yellowish red
Neutrophils - dark purple nuclei, pale pink cytoplasm, reddish-lilac small granules
Eosinophils - blue nuclei, pale pink cytoplasm, red to orange-red large granules
Basophils - purple to dark blue nucleus, dark purple, almost black large granules
Lymphocytes - dark purple to deep bluish purple nuclei, sky blue cytoplasm
Platelets - violet to purple granules
Parasites (Leishmania, malaria, etc.) - dark blue-black.

Leishman stain

0.15% Leishman powder in 100% methanol. Use after 24hrs.

Phosphate buffer (Sorensen)

Stock A:  0.2M sodium di-hydrogen orthophosphate (mw 156).
To prepare dissolve 3.12g in 100ml distilled water.

Stock B:  0.2M di-sodium hydrogen orthophosphate (mw 142).
To prepare dissolve 2.83g in 100ml ditilled water.

For pH 6.8 add 25.5ml of A to 24.5ml of B and make up to 100ml with distilled water.

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