ゴルジ法の一改良法について

A MODIFICATION OF GOLGI'S RAPID METHOD

I．はじめに

　Golgi法が発見されて100年余が経過したが，その染色結果の不安定さとはうらはらに，この方法によつて得られる所見の重要性は近来一段と増してきている。この不安定さをいささかでも除く為に我々はAldehyde系固定剤による灌流前固定の導入，固定及び鍍銀段階における液温の調節などに工夫を加え，ある程度の成果をあげ得たのでここに報告する。

As has been long known, the results of Golgi'srapid method are frequently uncertain from caseto case. In order to make the staining effect ofthis method highly stable, I introduced a newprecedure of perfusion-fixation using new aldehydefixatives.

The essential points of our method are as follows.(1) Young animals are preferable; age of the animalsused in this study: newborn or 4-week-old cats, 1-or 3-week-old rats, mice and rabbits. (2) Perfusionof the vascular system by using aldehyde fixatives(pH 7.2-7.4, Robertson's paraformaldehyde or 5%glutaraldehyde) under careful anesthesia withNembutal (Sodium pentobarbital). A cannula isplaced in the ascending aorta through an incisionmade in the left ventricle. An exit is providedby a small incision made in the right atrium. Theperfusion is carried out at low pressure (e. g., 1000mm H₂O) to avoid the destruction of the smallvessels. The perfusion volume is about 1ml pergr. of the body weight. (3) After the brain andspinal cord have been removed from the body ofthe animals, the material is cut into several pieces,preferably not more than 5-10 mm thick, and thenit is placed in a solution of the same aldehydefixative at room temperature for half a day (over-night) or longer, to a maximum of one month.After this, the blocks are rinsed in phosphate buffer(pH 7.2-7.4), with several changes, for one day.(4) The post-fixation is carried out in the followingsolution

2% O_sO₄………1 Vol.

3% K₂C_r2O₇………3-4 Vol.

for various periods of time depending on the sizeof block. The interrelation of thickness to thespeed of penetration of the osmium into the blockis indicated in Table I. The material should bekept at 18-23℃. (5) Silver-impregnation with 0.75% AgNO₃ for two days to one week, at 18-23℃.(6) Thick sections (100-150μ) are cut by microtomeafter rapid embedding, as follows: 70% alcohol→90% alcohol→100% alcohol (1 st and 2 nd)→absolutealcohol-ether→1/8 celloidin (celloidin 1 Vol.: abso-lute alcohol-ether 7 Vol.)→1/4 celloidin→1/2 celloi-din. The duration of each step is half a day (6-12hours). The embedding process should be completedwithin 7 days. In order to mount celloidin sectionssmoothly on the slides, Ramón-Moliner's methodis generally used.