Hexagonal array of subunits in intercellular junctions of the mouse heart and liver.
Hexagonal array of subunits in intercellular junctions of the mouse heart and liver.
复制标题
小鼠心脏和肝脏的细胞间连接处的亚基的六边形阵列。
DOI：
10.1083/jcb.33.3.c7
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发表时间：
1967-06
影响因子：
7.8
通讯作者：
Karnovsky, M J
中科院分区：
文献类型：
--
作者：
Revel, J P;Karnovsky, M J
材料和方法将组织固定在甲醛-戊二醛混合物 (6) 中，在缓冲液中洗涤，然后在 pH 7.2-7.4 的 OsO4-eolidine 中后固定 2 小时。在剧烈搅拌下，用 0.01 N NaOH 将 2-4% 硝酸镧溶液调节至 pH 7.6-7.8。碱的添加必须缓慢以避免氢氧化镧过早沉淀。当滴定正确完成时，在 pH 7.8 时会出现微弱的乳光或絮状物质的痕迹，在该 pH 值下氢氧化镧变得不溶。将等份的镧溶液添加到固定剂中，使得最终的镧浓度为0.5-1%。镧-四氧化锇-可力丁溶液的最终pH约为7.2并且微浑浊。当镧溶液达到高pH值时，似乎可能会形成一种胶体化合物，如下所述，该胶体化合物作为示踪剂渗透到细胞外空间。虽然我们认为最好将镧添加到直至醇的所有溶液中，但这不是必需的，并且可以单独在 OsO4 固定过程中添加镧。仅将镧添加到醛固定液或缓冲漂洗溶液中不太令人满意，因为在随后暴露于水溶液期间重金属似乎再次从块中被洗掉。暴露于镧溶液应在室温下进行。
MATERIALS AND METHODSTissues are fixed in a formaldehyde-glutaraldehyde mixture (6), are washed in buffer, and then are postfixed for 2 hr in OsO4-eollidine at pH 7.2-7.4. A 2-4% solution of lanthanum nitrate is brought to pH 7.6-7.8 with 0.01 N NaOH, with vigorous stirring. The addition of base must be made slowly to avoid premature precipitation of lanthanum hydroxide. When the titration is done properly, a faint opalescence or traces of flocculent material appear at pH 7.8, the pH at which lanthanum hydroxide becomes insoluble. An aliquot of the lanthanum solution is added to the fixatives so that the final lanthanum concentration is 0.5-1%. The final pH of the lanthanum-osmium tetroxide-collidine solution is about 7.2 and is faintly cloudy. It seems likely that, as lanthanum solutions are brought to a high pH, a colloidal compound is formed which, as described below, permeates the extracellular space as a tracer. While we deem it best to add lanthanum to all the solutions up to the alcohols, this is not necessary and lanthanum can be added during the OsO4 fixation alone. Adding the lanthanum only to the aldehyde fixing fluid or to the buffer rinse solution is less satisfactory, since the heavy metal seems to be washed out from the blocks again during subsequent exposure to aqueous solutions. The exposure to lanthanum solutions should be done at room temperature.