Fluorescence In Situ Hybridization on Tissue Sections
Fluorescence In Situ Hybridization on Tissue Sections
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DOI：
10.1007/978-1-4939-6703-2_11
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发表时间：
2017-01-01
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影响因子：
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通讯作者：
Lim, Tse Hui
中科院分区：
文献类型：
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作者：
Lim, Alvin S. T.;Lim, Tse Hui
福尔马林固定石蜡包埋（FFPE）组织通常是可用于实体组织（特别是肿瘤标本）FISH分析的标本。偶尔，从几个病人的组织微阵列构建的组织核心，同时进行评估。FFPE切片也可以从来源于细胞悬浮液的细胞块制备。间期荧光原位杂交试验采用靶向感兴趣的互补序列的特异性核酸序列（探针）来检测组织内基因/基因座或融合基因的获得或丢失。在本章中，我们描述了我们实验室使用的方案，包括载玻片脱蜡，预处理，蛋白酶处理，杂交，洗涤和复染。该方案可应用于所有早期FFPE制剂。通常，该测定需要连续3天才能完成，尽管可以进行更快速的测定。
Formalin-fixed paraffin-embedded (FFPE) tissues are typically the specimens available for FISH analysis of solid tissues, particularly of tumor specimens. Occasionally, tissue cores constructed as tissue microarrays from several patients are presented for simultaneous evaluation. FFPE sections can also be prepared from cell blocks derived from cell suspensions. The interphase fluorescence in situ hybridization assay employs specific nucleic acid sequences (probes) that target complementary sequences of interest to detect gains or losses of genes/gene loci or a fusion gene within the tissue. In this chapter, we describe the protocols utilized in our laboratory and include slide deparaffinization, pretreatment, protease treatment, hybridization, washing, and counterstaining. This protocol can be applied to all of the earlier FFPE preparations. In general, the assay takes 3 consecutive days to complete, although a more rapid assay can be performed.