各種神經節細胞内みとこんどりあノ形態學的研究 : 第1編 固定・包埋・染色法ニ關スル研究
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In order to improve the various old methods of fixing and staining mitochondria, I devised a method to colour most clearly the mitochondria in various ganglion cells. The method is as follows: 1) Inject the aorta of an animal with a moderate amount (according to the size of the animal) of 3% solution of kalium bichromat and formalin, (the ratio being parts of the former 100 to 15 of the latter). Draw the fluid out of the heart of the animal after the mixture has circulated through the whole body. 2) Take thin pieces out of the animal's cerebrum, cerebellum, spinal cord, sympathetic ganglion, spinal ganglion and accessory sympathetic ganglion. Immerse them a solution of kalium bichromat and formalin for one day if it is summer, for two days in winter, for a day and half in spring and autumn. 3) Leave them in a 2.5% sulution of kalium bichromat for three days in spring, summer and autumn; four days in winter. 4) Remove as much as possible of the water from the pieces with alcohol. Immerse them xylol, Put them in soft paraffine (melting point. 40℃) and then in hard paraffine (melting point. 58℃). Wrap them in the latter. 5) Prepare a saturant of acid fuchsin and a mixture made of pure anilin and distilled water. Put the saturant on the pieces which have been affixed to the object-glass. Warm them with a fire-flame twice in warm seasons or thrice in cold seasons. Wash them with water. 6) Dip them quickly in a solution of 0,3 aurantia in 100 alcohol. Wash them with water. 7) Immerse them in a saturate solution of light-green for live minutes. Wash them with water. 8) Benzin petrol→pure alcohol→xylol→balsam. In my experiments I found that when I used a weak acid fixing fluid, mitochondria in the ganglion cells became a strong colour; while when I used a strong acid fluid, they were coloured dark and their shapes changed from granular ones to bacillar. In these cases, the protoplasm of the ganglion cells became dark, yet mitochondria were not invisible. When I used a weak alkaline fixing fluid, the mitochondria were stained lightly and changed to fine granules with a decrease in the quantity; but wben I used a strong alkaline of the fluid, the mitochondria disappeared.
- 京都府立医科大学の論文
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