Combination of Gomori and Ziehl-Neelsen Stains for Showing Acid Phosphatase Activity andMycobacterium tuberculosisin Macrophages

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Abstract
Tubercle bacillus-infected macrophage monolayer cultures, after fixation in glutaraldehyde or without fixation, are stained by a Gomori method to show acid phosphatase activity. The method includes H,S to convert lead phosphate to sulphide. Only a minority of the mycobacteria is outlined by the black or brown stain; the similarity to the surrounding stained cytoplasmic particles makes identification difficult. The Gomori staining procedure is followed by a Ziehl-Neelsen method to stain acid-fast microorganisms; the temperature of the carbol fuchsin is just high enough to produce steaming, and the time of decolorisation is short. To avoid loss of the Gomori stain from the acid-fast procedure it is essential to repeat the exposure to H2S between the decolorisation and the light counterstaining. This combined method preserves the Gomori stain, against which the red acid-fast bacilli stand out sharply, so that acid phosphatase activity and bacteria can be located easily in the cell.

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