Brown-Hopps Method For Gram Positive And Gram Negative Bacteria
EMS Catalog #26106
10% Buffered Neutral Formalin
Paraffin, @ 6 microns
- Deparaffinize and hydrate to distilled water.
- Place slides on staining rack and pour on Crystal Violet Solution, 1%, Aqueous for two minutes. Rinse in distilled water.
- Mordant in Gram's Iodine for five minutes. Rinse in distilled water.
- Differentiate in Cellosolve, until blue color no longer streams away from the section (approximately 5-10 seconds). (See note)
- Quickly rinse in distilled water. (See note)
- Stain in Basic Fuchsin Solution, 0.5%, Aqueous for five minutes. Rinse in distilled water.
- Gallego's Solution , for five minutes. (Differentiates and "fixes" the Basic Fuchsin).
- Rinse thoroughly in distilled water and blot lightly to remove excess water (not to dryness).
- Stain for three seconds in Tartrazine Solution . Immediately blot away excess, but not to dryness).
- Cellosolve three changes, for six quick dips in each. (See note)
- Xylene, three changes, for 10 dips each. Slides may remain in Xylene until ready for mounting.
- Mount with Permount.
Gram Negative Bacteria
Gram Positive Bacteria
- Preferably, all stains and solutions are applied to the slide while it is in a horizontal position, except for steps 4, 10 and 11, where the slide should be dipped into the solutions contained in coplin jars.
- Steps 4, 5, 9 and 10 are critical with regard to timing.
- This procedure does not work well on Bouin's fixed tissues.
AFIP Manual of Histologic Staining Methods; ed. G. Luna, McGraw-Hill Publications, 3 rd . ed.; New York, c. 1968, p.224.
Brown, R. C. and Hopps, H.C., Geographic Pathology Division Armed Forces Institute Of Pathology, Washington, D.C. 20305
Crystal Violet Staining Solution 1% Aqueous, Gram Iodine Solution, Basic Fuchsin Staining Solution, Gallego’s Differentiating Solution, Cellosolve, and Tartrazine 1.5% Aqueous for Brown-Hopps Method For Gram Positive And Gram Negative Bacteria are available online from the EMS Catalog. For ordering or product information, click here.