福尔马林固定对DNA原位末端标记检测人脑细胞凋亡的影响。

The Histochemical Journal Pub Date : 1995-12-01
F D Davison, M Groves, F Scaravilli
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引用次数: 0

摘要

近年来,DNA原位末端标记(ISEL)技术已成为检测凋亡细胞的首选方法。将标记的核苷酸结合到内源性核酸酶切割的DNA单链区域以方便检测已被证明是一种敏感和直接的技术。先前的报道已将该技术应用于脑组织中凋亡细胞的研究,脑组织通常受到相对长期的福尔马林固定。在这项研究中,我们研究了长期福尔马林固定对各种病理和正常大鼠睾丸使用ISEL检测细胞凋亡能力的影响。在用福尔马林固定过夜的组织中,凋亡细胞很容易在可能发生的病理中被发现。然而,在固定数周或更长时间的组织中,未检测到凋亡细胞。对脑淋巴瘤组织和大鼠睾丸样本进行关于固定时间的前瞻性分析表明,检测凋亡细胞的能力在大约3-5周时减弱。为了避免假阴性结果的风险,最好在福尔马林固定时间少于这段时间的组织中使用ISEL。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The effects of formalin fixation on the detection of apoptosis in human brain by in situ end-labelling of DNA.

The technique of DNA in situ end-labelling (ISEL) for the detection of apoptotic cells has recently become the method of choice. The incorporation of a labelled nucleotide to facilitate detection into the single-stranded region of DNA cleaved by endogenous nucleases has proved to be a sensitive and straightforward technique. Previous reports have applied the technique to the study of apoptotic cells in brain tissue, which is normally subjected to relatively long-term formalin fixation. In this study we have examined the effects of long-term formalin fixation on the ability to detect apoptosis using ISEL in a variety of pathologies and in a normal rat testis. In the tissues which had been treated with overnight formalin fixation, apoptotic cells were readily identified in those pathologies where it might be expected to occur. However, in tissue which had been fixed for several weeks or more, apoptotic cells were not detectable. Samples of brain lymphoma tissue and rat testis subjected to a prospective analysis with respect to fixation time showed that the ability to detect apoptotic cells tailed off at around 3-5 weeks. In order to obviate the risk of false negative results it would be desirable to use ISEL in tissues formalin fixed for less than this period.

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