LEFFLERA METHODS ( F. Loeffler , it is mute. the bacteriologist, 1852 — 1915) — the methodical receptions at bacteriological researches developed and entered into practice by F. Leffler. From them in sovr, laboratory practice are applied: methods of preparation of the environment for cultivation of a diphtheritic stick — Leffler's circle (see. Diphtheria, etiology ), a way of coloring of diphtheritic bacteria and coloring of flagellums at bacteria.
The method of coloring of flagellums on Lefflera
Culture of bacteria is grown up on nutrient agar during 12 — 18 hours at an optimum temperature. Place 1,5 ml of the distilled or mains water in a pure test tube. A platinum loop take a small amount of the studied culture of bacteria and several times immerse it in water before receiving a uniform suspension. The suspension is put in the thermostat for 10 — 20 min. On the pure, calcinated slide plate, put with a platinum loop several drops of a suspension of culture of bacteria and dry up them on air, and then quickly fix, carrying out no more than 2 — 3 times through a flame of a torch. The fixed drug is immersed for 0,5 — 2 min. in pickle of the following structure: 20% water solution of a tannin — 100 ml; the solution of ferrous sulfate sated in the cold — 50 ml; saturated spirit solution of fuchsin — 10 ml. Processing in pickle is conducted at weak heating. After pickle drug is carefully washed out water and dried up, and then painted aniline fuchsin of Ehrlich within 2 — 3 min. Stained preparation is washed out water, dried up and mikroskopirut.
Modification of a method of Leffler according to M. A. Peshkov. Drugs are prepared on pure cover glasses. Pickle from three solutions is necessary for coloring.
1. Saturated spirit solution of the main fuchsin: add 10 ml of 96% of alcohol to 1 — 2 large crystals of fuchsin and put in the thermostat on 1 — 2 hour or on the water bath at t ° 50 °.
2. Water 20% solution of a tannin.
3. The solution of Mohr's salt (ferrous protoxidic ammoniac sulfate) sated in the cold. Mix 1 ml of solution 1 and 10 of ml of solution 2; to mix add 5,5 ml of solution 3; on the fixed drug pour the filtered solution of pickle much and leave it on glass for 10 — 15 min., watching that liquid did not dry up and did not leak under a lower surface of glass.
Drug is washed out a strong stream of a distilled water (30 sec.). Delete with a thin rag from edges of glass the remains of pickle and repeatedly wash out a distilled water. The processed drug a smear is placed in clock glass down and add the filtered Tsil's fuchsin diluted twice with water. Coloring is made within 3 — 5 min. then drug is washed out water.
Bibliography: The guide to microbiological diagnosis of infectious diseases, under the editorship of K. I. Matveev, page 34, M., 1973; Reference book on microbiological and virologic methods of a research, iod of an edition of M. O. Birger, page 28, M., 1973. See also bibliogr, to St. Leffler F .
D. M. Goldfarb.