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用于分析滑膜组织基因表达的激光介导显微切割技术。

Laser-mediated microdissection for analysis of gene expression in synovial tissue.

作者信息

Hashimoto Atsushi, Matsui Toshimichi, Tanaka Sumiaki, Ishikawa Akira, Endo Hirahito, Hirohata Shunsei, Kondo Hirobumi, Neumann Elena, Tarner Ingo Helmut, Müller-Ladner Ulf

机构信息

Department of Rheumatology and Infectious Diseases, Kitasato University School of Medicine, 1-15-1 Kitasato, Sagamihara 228-8555, Japan.

出版信息

Mod Rheumatol. 2007;17(3):185-90. doi: 10.1007/s10165-007-0564-0. Epub 2007 Jun 20.

Abstract

In experimental rheumatology, transcriptomics is one of the most important methods for investigating the pathogenesis of diseases. The biological material of most studies on rheumatoid arthritis has been bulk rheumatoid synovial tissues, but they are not suitable because they consist of several kinds of cells or structures. Laser-mediated microdissection (LMM) is a useful tool for isolating particular cells from tissue specimen to assess the functions of each cell. The LMM system employs a combination of a microscope and a laser-beam generator to cut out target areas on cryosections. Tissue compartments or even a single viable cell can be isolated using a non-focused laser beam without direct contact to avoid contamination, and this process is called laser pressure catapulting. An ultraviolet-A laser enables target cells to be procured without any influence on the surrounding. This technique has already been used in several studies in rheumatology, and its validity has been confirmed. Combined with other new techniques such as real-time quantitative polymerase chain reaction or microarray analysis, LMM is becoming more important in the analysis of gene expression in rheumatology.

摘要

在实验性风湿病学中,转录组学是研究疾病发病机制的最重要方法之一。大多数关于类风湿性关节炎研究的生物材料是大块的类风湿性滑膜组织,但它们并不适用,因为它们由多种细胞或结构组成。激光介导显微切割(LMM)是从组织标本中分离特定细胞以评估每个细胞功能的有用工具。LMM系统采用显微镜和激光束发生器的组合,在冷冻切片上切出目标区域。使用非聚焦激光束可以在不直接接触的情况下分离组织隔室甚至单个活细胞,以避免污染,这个过程称为激光压力弹射。紫外-A激光能够在不影响周围环境的情况下获取目标细胞。该技术已在多项风湿病学研究中得到应用,其有效性已得到证实。结合实时定量聚合酶链反应或微阵列分析等其他新技术,LMM在风湿病学基因表达分析中变得越来越重要。

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