From Big Medical Encyclopedia

BENINYETTI METHOD (Benignetti) — one of methods of coloring of flagellums of bacteria. In 2 — 3 days prior to use prepare solutions: I \solution of zinc sulfate (1 g) and a tannin (15 g) in a distilled water (100 ml); II \the aqueous solution of alum sated in a hot state; III \saturated spirit of a methylrosanilinum chloride. Before coloring solutions I, II, III mix in a proportion 5:5:3.

For coloring across Beninyetti use 18-hour bacterial culture which is grown up at an optimum temperature.

Drug for coloring is prepared as follows: having taken a loop of a little bacterial weight from the bottom of crops on a slant agar (but not from water of condensation), immerse a loop in a test tube from 5 ml of mains water until bacterial weight divorces. The drop of suspension is applied on pure clock glass and add a drop of 2% of aqueous solution of osmium tetroxide. After careful hashing mix is covered with other clock glass. The pure cover glass fixed by tweezers is slowly carried out by 3 — 4 times through an upper part of a flame of a torch. After glass cools down, a pasterovsky pipette consistently transfer to the surface processed by fire from clock glass five drops of suspension: one in the center, and four — on diagonals, are closer to corners. On the surface of well fat-free glass the put drops blur. When they dry, carry quickly out glass once through a flame (in case of the unsuccessful coloring of flagellums revealed at the subsequent microscopy it is necessary to try to do without this easy fixing).

On the fixed drug pour the freshly cooked filtered mix of solutions I, II, III, warm up 2 — 3 min. before emergence of vapors then drug is carefully washed out water, dry up, conclude in Canada balsam and investigate with immersion. See also Flagellums bacterial, Colouring of microorganisms.

M. A. Peshkov.