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激光显微切割技术在复杂组织中的应用。

Use of laser microdissection in complex tissue.

作者信息

Willenberg Holger S, Walters Rhodri, Bornstein Stefan R

机构信息

Department of Endocrinology, University of Düsseldorf, D-40225 Düsseldorf, Germany.

出版信息

Methods Enzymol. 2002;356:216-23. doi: 10.1016/s0076-6879(02)56935-4.

Abstract

Concomitant with the rapid development in biomedical knowledge, including the methods of molecular biology and proteomics, and the manufacture of ever more precise optical instruments, powerful lasers, and sophisticated microcomputing hardware and software, laser microdissection systems have emerged which are now entering the field of routine research. Today, several devices are commercially available, congresses devoted to the latest advances in laser microdissection are now held on regular occasions, and the number of publications based on the use of these techniques has risen to over 250. With laser microdissection, histological treatment, such as chemical or immunological fixation and staining, can readily be combined with methods suitable for molecular biology or proteomics. As the optical, technical, and methodological resolution of polymerase chain reaction (PCR) and microdissection increases, genetic and phenotypic studies of biological material are possible even at the level of single cells and subcellular elements. Moreover, questions such as the paracrine interaction of cells within complex tissues, the development of cancer, and the role of single cells in tissue remodeling or development on the microscopic and molecular level can now be addressed precisely at the molecular level. This chapter reviewed the development of laser microdissection platforms, its potential impact on the future of research, and how, in particular, these technologies can be successfully integrated into modern research and routine histopathological studies of complex tissue.

摘要

随着生物医学知识的迅速发展,包括分子生物学和蛋白质组学方法,以及越来越精密的光学仪器、强大的激光和先进的微型计算机硬件与软件的制造,激光显微切割系统应运而生,如今已进入常规研究领域。如今,有几种设备已商业化,定期举办专门探讨激光显微切割最新进展的会议,基于这些技术的出版物数量已增至250多篇。借助激光显微切割,组织学处理,如化学或免疫固定及染色,可轻松与适合分子生物学或蛋白质组学的方法相结合。随着聚合酶链反应(PCR)和显微切割在光学、技术和方法上分辨率的提高,即使在单细胞和亚细胞水平对生物材料进行基因和表型研究也成为可能。此外,诸如复杂组织内细胞的旁分泌相互作用、癌症的发展以及单细胞在微观和分子水平上在组织重塑或发育中的作用等问题,现在都可以在分子水平上得到精确解答。本章回顾了激光显微切割平台的发展、其对未来研究的潜在影响,以及这些技术如何尤其能成功整合到复杂组织的现代研究和常规组织病理学研究中。

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