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使用胶带法和手动芯片点样仪手动构建组织微阵列。

Manual Construction of a Tissue Microarray using the Tape Method and a Handheld Microarrayer.

机构信息

Department of Research, Mayo Clinic.

Department of Laboratory Medicine and Pathology, Mayo Clinic.

出版信息

J Vis Exp. 2022 Jun 10(184). doi: 10.3791/63086.

DOI:10.3791/63086
PMID:35758657
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10448997/
Abstract

The tissue microarray (TMA) is an important research tool in which many formalin fixed paraffin embedded (FFPE) samples can be represented in a single paraffin block. This is achieved by using tissue cores extracted from the region of interest of different donor FFPE blocks and arranging them into a single TMA paraffin block. Once constructed, sections from the completed TMA can be used to perform immunohistochemistry, chromogenic, fluorescence in situ hybridization (FISH) and RNA ISH studies to assess protein expression as well as genomic and transcriptional alterations in many samples simultaneously, thus minimizing tissue usage and reducing reagent costs. There are several different TMA construction techniques. One of the most common construction methods is the recipient method, which works best with cores of the same length for which a minimum length of 4 mm is recommended. Unfortunately, tissue blocks can be heavily resected during the diagnostic process, frequently resulting in "non-ideal" donor block thicknesses of less than 4 mm. The current article and video focus on the double-sided adhesive tape method; an alternative manual, low cost, easy to use, and rapid method to construct low density (<50 cores) TMAs that is highly compatible with these non-ideal donor blocks. This protocol provides a step-by-step guide on how to construct a TMA using this method, with a focus on the critical importance of pathological review and post construction validation.

摘要

组织微阵列(TMA)是一种重要的研究工具,它可以在单个石蜡块中代表许多福尔马林固定石蜡包埋(FFPE)样本。这是通过使用从不同供体 FFPE 块的感兴趣区域提取的组织芯,并将它们排列到单个 TMA 石蜡块中实现的。一旦构建完成,从完整的 TMA 中切取的切片可用于进行免疫组织化学、显色、荧光原位杂交(FISH)和 RNA ISH 研究,以同时评估许多样本中的蛋白质表达以及基因组和转录改变,从而最大限度地减少组织使用并降低试剂成本。有几种不同的 TMA 构建技术。最常见的构建方法之一是接收方方法,该方法最适用于长度相同的芯,建议最小长度为 4 毫米。不幸的是,在诊断过程中组织块可能会被大量切除,经常导致供体块厚度小于 4 毫米的“不理想”情况。本文和视频重点介绍双面胶带法;这是一种替代的手动、低成本、易于使用且快速的方法,可构建低密度(<50 个芯)TMA,与这些非理想的供体块高度兼容。该方案提供了使用该方法构建 TMA 的分步指南,重点介绍病理审查和构建后验证的重要性。