Giemsa Stain Procedure

The procedures for staining thick and thin films differ. Staining is usually done in a Coplin jar. The stain must be made fresh each day. 

Stain slides with only a thin film as follows. 

Prepare a 1 40 dilution of stock Giemsa stain in neutral buffered water, pH 7.0 to 7.2 (generally, 2 ml of Giemsa stock plus 38 ml of buffered water with 0.01% Triton X-100). 

Stain the film for approximately 60 min (the time, which will vary slightly with different lots of stock Giemsa stain, can be determined by the staining of leukocytes and erythrocytes). 

Wash the slide briefly by dipping it in buffered water. 

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Thick and thin films are best stained with Giemsa stain, as it provides the most detailed and intense staining of parasites. Wright stain can be used for thin films but not for thick films, as it contains alcohol, which will fix the erythrocytes. Wright stain does not stain parasites as well as Giemsa stain. The staining procedure is outlined below. 

Much of the confusion was caused by the inadequacy of the staining procedures. By means of the HCl-Giemsa staining technique of Piekarski, many observations have been reported demonstrating the presence of chromatinic structures in bacteria. 

Staining can be achieved by standard procedures utilizing acetic-orcein, Giemsa stain, or the Feulgen reaction. 

For measuring plating efficiency a similar procedure is adopted and the colonies are fixed and stained (with Giemsa) and their number compared with the initial input. The plating efficiency of primary cells is low, but plating efficiencies of up to 100% can be achieved with cell lines or cell strains. 

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