The autometallographic zinc-sulphide method. A new approach involving in vivo creation of nanometer-sized zinc sulphide crystal lattices in zinc-enriched synaptic and secretory vesicles
The autometallographic zinc-sulphide method. A new approach involving in vivo creation of nanometer-sized zinc sulphide crystal lattices in zinc-enriched synaptic and secretory vesicles
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DOI：
10.1007/bf02331399
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发表时间：
1996-05-01
期刊：
影响因子：
--
通讯作者：
Danscher, G
中科院分区：
文献类型：
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作者：
Danscher, G
提出了蒂姆硫化银方法的新版本，涉及富锌末端中锌离子的体内结合。通过将硫化钠注射到深度麻醉动物的腔静脉中，可以以化学方式结合分泌囊泡和突触囊泡中的囊泡锌，即可螯合锌（锌离子），以硫化锌晶格的形式。静脉注射后四分钟，用戊二醛、戊二醛和甲醛的磷酸盐缓冲溶液或盐水溶液对动物进行经心灌注。然后，通过暴露于自动金相显影剂，可以在低温恒温器和振动切片机部分中对纳米尺寸的催化晶体进行银放大。结果表明，在银增强的同时，硫化锌晶体转变成相应的硫化银晶体。对于超微结构研究，建议使用振动切片机切片进行自动金相显影。从这些切片中，从感兴趣的区域切下组织块，用四氧化锇进行块染色并嵌入 Epon 中。这种方法可以对切片进行锌特异性自动金相染色，具有前所未有的高技术质量。
A new version of Timm's sulphide silver method involving in vivo binding of zinc ions in zinc enriched terminals is presented. By injecting sodium sulphide into the vena cava of deeply anaesthetized animals, it is possible to bind chemically the vesicular zinc, i.e. chelatable zinc (zinc ions), in secretory and synaptic vesicles, in the form of zinc sulphide crystal lattices. Four minutes after the intravenous injection the animal is perfused transcardially with a phosphate-buffered solution of glutaraldehyde, glutaraldehyde and formaldehyde, or with a saline solution. The nanometer-sized catalytic crystals can then be silver-amplified in cryostat and vibratome sections by exposure to an autometallographic developer. It is demonstrated that contemporaneously with silver enhancement, the zinc sulphide crystals are transformed to the corresponding silver sulphide crystals. For ultrastructural studies, autometallographic development of vibratome sections is recommended From these sections tissue blocks are cut from the areas of interest, blockstained with osmium tetroxide and embedded in Epon. This approach results in a zinc-specific autometallographic staining of the sections of a hitherto unseen, high technical quality.