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用于实时聚合酶链反应样本制备的激光辅助显微切割

Laser-assisted microdissection for real-time PCR sample preparation.

作者信息

Pinzani P, Orlando C, Pazzagli M

机构信息

Department of Clinical Physiopathology, Clinical Biochemistry Unit, University of Florence, Italy.

出版信息

Mol Aspects Med. 2006 Apr-Jun;27(2-3):140-59. doi: 10.1016/j.mam.2005.12.006. Epub 2006 Feb 9.

Abstract

Laser-assisted microdissection (LMD) has been developed to procure precisely the cells of interest in a tissue specimen, in a rapid and practical manner. Together with real-time PCR and RT-PCR techniques, it is now feasible to study genetic alterations, gene expression features and proteins in defined cell populations from complex normal and diseased tissues. The process that brings from sample collection to the final quantitative results is articulated in several steps, each of which requires optimal choices in order to end up with high-quality nucleic acid or protein that allows successful application of the final quantitative assays. This review will describe shortly the development of LMD technologies and the principles they are based on. Trying to highlight the advantages and disadvantages of LMD, the main problems related to specimens collection and processing, section preparation and extraction of bio-molecules from microdissected tissue samples have been analysed.

摘要

激光辅助显微切割(LMD)技术已被开发出来,可快速、实用地从组织样本中精确获取感兴趣的细胞。结合实时PCR和RT-PCR技术,现在可以研究复杂的正常组织和病变组织中特定细胞群体的基因改变、基因表达特征和蛋白质。从样本采集到最终定量结果的过程分为几个步骤,每个步骤都需要做出最佳选择,以便最终获得高质量的核酸或蛋白质,从而成功应用最终的定量分析。本综述将简要描述LMD技术的发展及其所基于的原理。通过分析LMD的优缺点,重点探讨了与标本采集和处理、切片制备以及从显微切割组织样本中提取生物分子相关的主要问题。

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