组织切片中电子致密的人工沉积物:乙醇、醋酸铀酰和磷酸盐缓冲液的作用。

J Louw, K Williams, I S Harper, S A Walfe-Coote
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引用次数: 11

摘要

在制备用于透射电子显微镜的乙醛固定组织切片中出现电子致密沉积物归因于许多相互冲突的因素。为了澄清这一点,在试管中研究了磷酸盐或羧酸缓冲液、戊二醛、乙醇和醋酸铀酰的不同组合的沉淀效果。作为初步研究,研究了磷酸缓冲液、乙醇和醋酸铀酰在戊二醛固定的心脏和肾脏组织中的联合作用。在这些组织中形成电子致密沉积物的基本因素似乎是磷酸盐缓冲液、乙醇和醋酸铀酰,尽管戊二醛可能以某种方式起作用。矿床的性质和强度似乎随这些因素组合的顺序而变化。锇似乎不是反应的必要因素,因为在浸渍和未浸渍的组织中都观察到沉积。为避免此类沉积,如果使用磷酸盐缓冲液作为固定载体或在初始固定液后进行缓冲洗涤,建议在熏蒸后蒸馏水洗涤20 - 30分钟,然后用醋酸铀酰水进行整体染色。
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Electron dense artefactual deposits in tissue sections: the role of ethanol, uranyl acetate and phosphate buffer.

The occurrence of electron dense deposits in sections of aldehyde-fixed tissue prepared for transmission electron microscopy has been attributed to a number of conflicting factors. In an attempt to clarify this, the precipitating effect of different combinations of phosphate or cacodylate buffer, glutaraldehyde, ethanol and uranyl acetate was investigated in test tubes. As a preliminary investigation the combination of phosphate buffer, ethanol and uranyl acetate was investigated in heart and kidney tissue fixed in glutaraldehyde with or without postosmication. The essential factors in the formation of electron dense deposits in these tissues appear to be phosphate buffer, ethanol, and uranyl acetate, although glutaraldehyde may contribute in some way. The nature and intensity of the deposits seem to vary with the sequence of combination of these factors. Osmium did not appear to be an essential factor in the reaction since deposits were observed in both osmicated and unosmicated tissue. To avoid such deposits, a postosmication distilled water wash for 20 to 30 min followed by en bloc staining with aqueous uranyl acetate is advised if phosphate buffer is used as a fixative vehicle or buffer wash after the primary fixative.

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