Mitochondria in tissue culture
Mitochondria in tissue culture
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DOI：
10.1126/science.39.1000.330
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发表时间：
1914-01-01
期刊：
影响因子：
56.9
通讯作者：
Lewis, WH
中科院分区：
文献类型：
--
作者：
Lewis, MR;Lewis, WH
雅努斯绿色的优势。00001和。000005的1%将活细胞中的线粒体染成亮蓝绿色。然而，颜色会在15分钟到3小时内褪色，我们无法重新染色。janus绿色也有轻微的毒性，在几个小时内杀死细胞。我非常感谢E博士。V.Cowdry从芝加哥的Bensley博士那里获得了这种特殊的janus绿色（二乙基藜芦胺偶氮二甲基苯胺）。尝试用另一种品牌染色，但未成功。没吹B。我们感谢赫伯特·埃文斯博士，他用的是非常稀的。000005到。0000025用于检测与线粒体连接的脂质。它将类脂染成粉红色，但不幸的是有点毒性，并且像janus绿色一样，在几小时内杀死细胞。通过将盖玻片置于锇酸蒸汽室中2至5分钟来固定制备物，由于生长非常薄，固定几乎是瞬间的，并且线粒体在形态上与活细胞相同。在硬化过程中，通过在70%的酒精中滴几滴过氧化氢来漂白由渗透引起的变黑，然后用海登海因铁苏木素对制剂进行染色。由于在某些地方，细胞在盖玻片的下表面上被压平成比单个细胞的通常厚度薄得多的单层，人们可以用最小的聚焦量来研究整个活细胞及其内容物。此外，在观察期间的任何时刻，可以固定培养物，随后在染色制剂中研究相同的细胞。
Janus green in strengths of. 00001 and. 000005 of 1 per cent. stains the mitochondria in the living cells a brilliant blue green. The color fades, however, in from 15 minutes to 3 hours, and we have been unable to restain. The janus green is also slightly toxic and kills the cells in a few hours. We are indebted to Dr. E. V. Cowdry for this particular janus green (di ethyl saffranin azo di methyl aniline) which was obtained by him from Dr. Bensley of Chicago. Attempts to stain with another make were unsuccessful. Nilblew B. extra, for which we are indebted to Dr. Herbert Evans, was used in very dilute solution of. 000005 to. 0000025 of 1 per cent. for the detection of lipoids in con-nection with the mitochondria. It stains the lipoids pink, but is unfortunately somewhat toxic and, like the janus green, kills the cells in a few hours.The preparations are fixed by placing the coverslip in a chamber of osmic acid vapor from two to five minutes and since the growth is very thin, the fixation is almost instantane-ous and the mitochondria remainspractically the same as in the living cells. The blackening caused by the osmic is bleached during the hardening processes by means of a few drops of hydrogen peroxide in the 70 per cent. alcohol and the preparations are then stained with Heidenhain's iron hematoxylin. Since in places the cells are flattened out on the under surface of the coverslip into a single layer much thinner than the usual thickness of a single cell, one can study the entire living cell and its contents with a minimal amount of focusing. Also at any moment during the observations the culture can be fixed and later the same cells studied in a stained prep-aration.