SNESAREVA METHODS ( P.E. Snesarev , the Soviet neuromorphologist, 1876 — 1954) — methods of coloring of drugs for identification of elements of a neuroglia of tissue of brain and argyrophil fibers of connecting fabric.
During the coloring by this method use the material fixed in 15 — 20% neutral formalin. For complex coloring of a neuroglia frozen sections 5 — 20 microns thick on 15 — 20 sec. immerse water solution of erythrosine in 1%, wash out a distilled water and on 30 sec. transfer to 0,5% solution phosphatomolybdic to - you with addition of solution of bicarbonate potassium (1:1000) — 5 — 10 drops on 20 ml to - you. The cut is rinsed with a distilled water, spread on the glass greased protein-glycerin and processed mix of methyl alcohol with chloroform (1:2) to an enlightenment. On drops apply Mai's solution on drug — Gryunvalda, to-ry in 30 — 40 sec. part with 1 — 3 drops of a distilled water, continuing coloring of 5 — 10 more min. Then paint is merged, the cut is rinsed with a distilled water and processed for the purpose of a differentiation divorced ice acetic to - that (1:2) before accurate identification of gray and white matter of a brain. Drug is washed out a distilled water, dried filter paper, immersed on 1 — 3 sec. in acetone, out through a xylol and concluded in balm. Glial fibers are painted in blue color, kernels of cells — in pink and blue, well comes to light patol. granularity of a neuroglia, a myelin gains reddish color, Negri's little bodies — blue.
For impregnation of argyrophil fibers of connecting fabric well washed out pieces of fabric maintain in the thermostat at f 37 ° from 6 to 48 hours at first in 12%, and then in 24% solution of gelatin for 1% to carbolic water. Gelatin is quickly cooled before hardening, cut out from it blocks with pieces of fabric, condense blocks in 10% formalin, quickly washed out and cut on the freezing microtome. Cuts 5 — 20 microns thick on 48 hours place solution of feather alum (solution of ammonium-iron of sulfate) in 4%, wash out in 2 — 3 portions of a distilled water, postpone to 24 hours in 5 — 10% solution of silver nitrate (in the dark) and on one impregnate ammoniac silver of 5 — 10 min. (see. Bilshovsky — Groce — Lavrentyeva a method ). Further cuts process within 3 — 5 min. 20% solution of neutral formalin, wash out in mains water, gild (see. Gilding methods ), fix 5% solution of hyposulphite, dokrashivat kernels hematoxylin or a carmine, dehydrate and conclude in balm. Argyrophil fibers (see) are painted in dark brown color, and a surrounding background — in yellow. The method is suitable also for the paraffin sections pasted on glass; in this case cuts before treatment by gelatin dewax by means of organic solvents and carefully wash out a distilled water.
Besides, the modification of coloring of nervous cells by Nigel's method intended for coloring of cuts of a brain belongs to S. of m (see. Nisslya method ), modification of pan-optical coloring across Pappengeym (see. Pappengeyma methods ) and modification of a method of Altmann of the coloring of mitochondrions suitable for the material fixed by formalin (see. Altmanna methods ).
Bibliography: Merkulov G. A. Course of the patologogistologichesky equipment, page 187, L., 1969; Snesarev P. E. About use of silver sedimentary methods to paraffin series, Laborat. prakt., No. 9, page 1, 1932.
Ya. E. Hesinonim