Brown-Brenn 100 tests

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Product for the preparation of cyto-histological samples for optical microscopy. For demonstration Gram-positive and Gram-negative bacteria in tissue sections and smears.

Gram staining is the most important method to differentiate bacteria species. Two dyes are used one after the other: crystal violet and fuchsine. Crystal violet solution precipitates through oxidation with a iodine solution. The deriving complex attaches to bacteria cell walls with bonds of varying nature and intensity. The differentiating solution removes the crystal violet-iodine complex from the walls of some bacteria, but it does not act on others. These retain the primary dye and are called Grampositive. Decolorized bacteria are then counterstained with a red dye; they are called Gram-negative. Gram-positive bacteria’s capacity to retain the dye-iodine complex is usually ascribed to the bond which develops between the complex and a molecule only Gram-positive possess, namely magnesium ribonucleate.

METHOD:
1) Bring section to distilled water.
2) Put on the section 8 drops of reagent A and 2 drops of reagent B: leave to act 1 minute.
3) Wash in distilled water.
4) Put on the section 10 drops of reagent C: leave to act 3 minutes.
5) Rinse in water and blot with filter paper to complete dryness.
6) Put on the section 10 drops of reagent D: leave to act 1 minute.
7) Drain the slide without washing.
8) Put on the section 10 drops of reagent E: leave to act 1 minute.
9) Rinse in water and blot with filter paper.
10) Put on the section 10 drops of reagent F: leave to act 1 minute.
11) Drain the slide without washing.
12) Put on the section 10 drops of reagent G: leave to act 30 seconds.
13) Clear in xylene and mount.