Comparative usefulness of tissue fixatives for in situ viral nucleic acid hybridization.
Comparative usefulness of tissue fixatives for in situ viral nucleic acid hybridization.
复制标题
组织固定剂在原位病毒核酸杂交中的比较有用性。
DOI：
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复制
发表时间：
1985
影响因子：
3.2
通讯作者：
D. Rock
中科院分区：
文献类型：
--
作者：
H. McAllister;D. Rock
传统上，用于病毒核酸原位杂交的组织是从实验室啮齿动物获得的小块，专为电子显微镜设计的固定剂，例如高碘酸盐-赖氨酸-多聚甲醛（PLP）可以充分处理它们。然而，这些固定剂的渗透能力有限，并且可能不会产生明显的硬化，因此评估了替代的固定方法。目的是确定适合大体积组织（例如整个或半个猪脑和牛脑）的固定剂是否也与原位杂交兼容。使用伪狂犬病病毒 (PRV) 颅内接种 BALB/c 小鼠的系统，然后用 35S 标记的 PRV DNA 探针对脑组织切片进行原位杂交，对各种固定剂进行了评估。组织切片的丢失是一个主要问题，特别是对于 PLP 和福尔马林，但测试的五种固定剂获得了阳性结果。使用 PLP 和改良卡诺氏液 MOCA 固定剂时，细胞形态尤其良好。一个偶然但重要的观察结果是福尔马林与原位杂交兼容。因此，使用常规处理的组织块（福尔马林固定、石蜡包埋）对病毒性疾病进行回顾性研究是可以想象的。
Traditionally tissues for in situ hybridization of viral nucleic acid have been small pieces obtained from laboratory rodents, and fixatives that are designed for electron microscopy, such as periodate-lysine-paraformaldehyde (PLP) can handle them adequately. However, these fixatives have limited penetrating ability and may produce no appreciable hardening, so alternative fixation methods were evaluated. The intention was to determine whether fixatives adequate for bulky tissues such as whole or halved pig and cow brains would also be compatible with in situ hybridization. Various fixatives were evaluated using a system of intracranial inoculation of BALB/c mice with pseudorabies virus (PRV) followed by in situ hybridization of brain tissue sections with a 35S-labeled PRV DNA probe. Loss of tissue sections was a major problem, particularly with PLP and formalin, but positive results were obtained with five fixatives tested. Cellular morphology was especially good with PLP and with a modification of Carnoy's fluid, MOCA fixative. An incidental but important observation was that formalin is compatible with in situ hybridization. Retroactive studies of viral diseases using routinely processed blocks of tissue (formalin-fixed, paraffin-embedded) are therefore conceivable.
通过原位杂交在急性和潜伏感染期间将单纯疱疹病毒定位在小鼠中枢神经系统的三叉神经和嗅觉系统中。
DOI：
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发表时间：
1984
期刊：
Laboratory investigation; a journal of technical methods and pathology
影响因子：
--
作者：
Stroop,WG;Rock,DL;Fraser,NW
通讯作者：
Fraser,NW