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一种用于生物样本的冷冻断裂复型装置。

A freeze-fracture replication apparatus for biological specimens.

作者信息

Stolinski C

出版信息

J Microsc. 1975 Aug;104(3):235-44. doi: 10.1111/j.1365-2818.1975.tb04021.x.

Abstract

A freeze-fracture apparatus of original design has been constructed which can be fitted onto a standard vacuum evaporator unit. In it, cell suspensions and organized tissue may be processed by inserting a sample into a cylindrical holder. By leaving a small part of the tissue protruding from the holder, pre-selected and aligned portions of the specimen can subsequently be revealed by fracture under vacuum. After rapid freezing, the specimen remains firmly attached to the inner wall of the sample holder, preventing its possible loss during fracturing. A mechanism, in the form of a double-sided converging wedge, which is operated from outside the vacuum chamber, is used to produce a fracture in the specimen. The device gently induces a fracture in the desired part of the tissue and lifts the protruding part of the specimen out of the way. In this way, reasonably flat fracture faces are produced for subsequent replication. As the fracturing mechanism comes into contact only with the outer edges of the specimen, damage and contamination liable to occur when the entire specimen is traversed by a blade, is avoided. In addition the specimen stage is surrounded by a cold metal shroud which acts as an efficient trap for contaminants. In this way, favourable vacuum conditions are produced in the vicinity of the specimen. Such effective enclosing of the specimen also facilitates controlled sublimation of the sample.

摘要

已构建了一种原始设计的冷冻断裂装置,该装置可安装在标准真空蒸发器单元上。在该装置中,通过将样品插入圆柱形支架中,可以对细胞悬液和有组织的组织进行处理。通过使一小部分组织从支架中突出,随后可以通过在真空中断裂来显示标本预先选择并对齐的部分。快速冷冻后,标本仍牢固地附着在样品支架的内壁上,防止其在断裂过程中可能丢失。一种以双面会聚楔形式存在的机构,从真空室外部操作,用于在标本中产生断裂。该装置在组织的所需部位轻轻引发断裂,并将标本的突出部分移开。通过这种方式,可以产生相当平坦的断裂面以供后续复制。由于断裂机构仅与标本的外边缘接触,避免了在整个标本被刀片穿过时容易发生的损坏和污染。此外,样品台被一个冷金属罩包围,该冷金属罩可作为污染物的有效捕集器。通过这种方式,在标本附近产生了有利的真空条件。对标本的这种有效封闭也有助于样品的可控升华。

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