Staining Method of Hotchkiss

Staining Methods in Microbiology
Part 1
Nuclear Material Stains
Staining Methods of Robinow
Rapid Method of Robinow
Feulgen Stain
Staining with Acridine Oragne
Metachromatic Granule (Volutin) Stains
Staining with Methylene Blue
Staining Method of Gohar
Staining Method of Albert Modified by Laybourn
Stain For Intracellular Lipid
Staining Method of Burdon
Stain For Polysaccharides
Staining Method of Hotchkiss
Alcian Blue Stain
Special Staining Techniques
Spirochete Stains
Staining Method of Fontana
Staining Method of Rue
Staining Method of Lavadi
Stain For Azotobactor Cysts
Stains For Mycoplasmas
Staining Method of Diens
Staining of Colony
Coverslip preparation
Staining with Cresyl Fast Violet

Staining Method of Hotchkiss

	Staining Method of Hotchkiss Polysaccharide is oxidized with period ate to form polyaldehyde which reacts with Schiff's fuchsin sulphite to give rise to red colour. The proteins and nucleic acids remain colourless. The method may be used to reveal fungal elements in sections of infected animal tissue where fungi is stained red, while the tissue material, except glycogen and mucin, fail to take up stain.
The procedure for staining is as follows. Prepare a smear and fix it with heat. Flood the slide with periodate solution (S47) and allow it to react for 5 minutes. Rinse the smear with 70% ethyl alcohol. Cover the smear with reducing rinse solution (S48) and keep it for 5 minutes. Rinse the smear, with 70% ethyl alcohol. Stain the smear with fuchsin-sulphite solution (S49) for 15-45 minutes. Wash the smear several times with freshly prepared sulphite wash solution (S50) and then with water. Counterstain with 0.002% of malachite green for few seconds. Wash, blot dry and examine under oil-immersion objective. Polysaccharides will be stained red in contrast to the green cell.
This is a simple stain for polysaccharides and gives satisfactory results. The critical step in this procedure is the counter staining. Excess exposure will overstain the preparation. Following schedule for staining is recommended (12). Prepare the smear and heat fix in the usual way. Stain the smear by allowing it to react with alcian blue (S51) for 1 minute. Wash with water and air dry the smear.
Counter stain the preparation with 1:9 diluted carbo1 fuchsin (S19) and wash off immediately with water. Air dry and examine under oil-immersion objective. Capsules and other bacterial polysaccharides will appear blue, while cellular material will appear red in colour.

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