Datta Soma, Malhotra Lavina, Dickerson Ryan, Chaffee Scott, Sen Chandan K, Roy Sashwati
Department of Surgery, Center for Regenerative Medicine and Cell Based Therapies and Comprehensive Wound Center, Laser Capture Molecular Core, The Ohio State University Wexner Medical Center, Columbus, Ohio, USA.
Histol Histopathol. 2015 Nov;30(11):1255-69. doi: 10.14670/HH-11-622. Epub 2015 Apr 20.
Any tissue is made up of a heterogeneous mix of spatially distributed cell types. In response to any (patho) physiological cue, responses of each cell type in any given tissue may be unique and cannot be homogenized across cell-types and spatial co-ordinates. For example, in response to myocardial infarction, on one hand myocytes and fibroblasts of the heart tissue respond differently. On the other hand, myocytes in the infarct core respond differently compared to those in the peri-infarct zone. Therefore, isolation of pure targeted cells is an important and essential step for the molecular analysis of cells involved in the progression of disease. Laser capture microdissection (LCM) is powerful to obtain a pure targeted cell subgroup, or even a single cell, quickly and precisely under the microscope, successfully tackling the problem of tissue heterogeneity in molecular analysis. This review presents an overview of LCM technology, the principles, advantages and limitations and its down-stream applications in the fields of proteomics, genomics and transcriptomics. With powerful technologies and appropriate applications, this technique provides unprecedented insights into cell biology from cells grown in their natural tissue habitat as opposed to those cultured in artificial petri dish conditions.
任何组织都是由空间分布的细胞类型组成的异质混合物。响应任何(病理)生理信号,任何给定组织中每种细胞类型的反应可能都是独特的,无法在细胞类型和空间坐标上进行同质化处理。例如,在心肌梗死的情况下,一方面心脏组织中的心肌细胞和成纤维细胞反应不同。另一方面,梗死核心区的心肌细胞与梗死周边区的心肌细胞反应也不同。因此,分离纯的目标细胞是对参与疾病进展的细胞进行分子分析的重要且必要步骤。激光捕获显微切割(LCM)能够在显微镜下快速、精确地获得纯的目标细胞亚群,甚至单个细胞,成功解决了分子分析中组织异质性的问题。本综述概述了LCM技术、其原理、优点和局限性以及在蛋白质组学、基因组学和转录组学领域的下游应用。借助强大的技术和适当的应用,该技术提供了前所未有的对细胞生物学的见解,这些见解来自于在其自然组织环境中生长的细胞,而非在人工培养皿条件下培养的细胞。