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来自冷冻组织的组织微阵列-OCT技术。

Tissue microarrays from frozen tissues-OCT technique.

作者信息

Fejzo Marlena Schoenberg, Slamon Dennis J

机构信息

Division of Hematology/Oncology, Department of Medicine, School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA.

出版信息

Methods Mol Biol. 2010;664:73-80. doi: 10.1007/978-1-60761-806-5_8.

Abstract

Tissue microarray technology is a new method used to analyze 100s to 1,000s of tumor samples on a single slide allowing high throughput analysis of genes and proteins on a large cohort. The original methodology involves coring tissues from paraffin-embedded tissue donor blocks and placing them into a single paraffin block. One difficulty with paraffin-embedded tissue relates to antigenic changes in proteins and mRNA degradation induced by the fixation and embedding process. We modified this technology by using frozen tissues embedded in OCT compound as donor samples and arraying the specimens into a recipient OCT block. Tumor tissue is not fixed before embedding, and sections from the array are evaluated without fixation or postfixed according to the appropriate methodology used to analyze a specific gene at the DNA, RNA, and/or protein levels. While paraffin tissue arrays can be problematic for immunohistochemistry and for RNA in situ hybridization analyses, this method allows optimal evaluation by each technique and uniform fixation across the array panel. Frozen tissue microarrays work well for DNA, RNA, and protein analyses, and may have significant advantages over the original technology for the assessment of some genes and proteins by improving both qualitative and quantitative results.

摘要

组织微阵列技术是一种新方法,可在一张载玻片上分析数百至数千个肿瘤样本,从而对大量队列中的基因和蛋白质进行高通量分析。原始方法包括从石蜡包埋的组织供体块中取芯组织,并将它们放入单个石蜡块中。石蜡包埋组织的一个难题与固定和包埋过程引起的蛋白质抗原性变化和mRNA降解有关。我们通过使用包埋在OCT复合物中的冷冻组织作为供体样本,并将标本排列到受体OCT块中,对该技术进行了改进。肿瘤组织在包埋前不固定,根据用于在DNA、RNA和/或蛋白质水平分析特定基因的适当方法,对阵列切片进行评估时不进行固定或后固定。虽然石蜡组织阵列在免疫组织化学和RNA原位杂交分析中可能存在问题,但这种方法允许通过每种技术进行最佳评估,并在整个阵列板上实现均匀固定。冷冻组织微阵列在DNA、RNA和蛋白质分析中效果良好,并且通过改善定性和定量结果,在评估某些基因和蛋白质方面可能比原始技术具有显著优势。

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