Hoffmann Julia, Wilhelm Jochen, Kwapiszewska Grazyna
Ludwig Boltzmann Institute for Lung Vascular Research, Graz, Austria.
Department of Internal Medicine, Justus-Liebig-University, Giessen, Germany.
Methods Mol Biol. 2017;1627:325-340. doi: 10.1007/978-1-4939-7113-8_21.
The heterogeneous nature of most human organs and tissues represents a common challenge when analyzing specific structures or cells. Laser capture microdissection (LCM) enables isolation of pure cells from a mixed population of cells or tissue samples via usage of laser energy. Combined with high-throughput gene or protein techniques, compartment specific analysis elucidating the role of specialized cell types in physiological or pathophysiological activity can be performed. This chapter describes the crucial steps that have to be taken into consideration when designing and conducting a LCM project. Detailed protocols describing the workflow from project planning to high-throughput analysis of LCM material used in our laboratory are provided. Routinely occurring challenges and appropriate solutions, e.g., when working with fibrotic tissue are described.
在分析特定结构或细胞时，大多数人体器官和组织的异质性是一个常见的挑战。激光捕获显微切割（LCM）通过使用激光能量，能够从混合的细胞群体或组织样本中分离出纯细胞。结合高通量基因或蛋白质技术，可以进行隔室特异性分析，以阐明特定细胞类型在生理或病理生理活动中的作用。本章描述了在设计和实施LCM项目时必须考虑的关键步骤。提供了详细的方案，描述了从项目规划到我们实验室使用的LCM材料的高通量分析的工作流程。还描述了常规出现的挑战和适当的解决方案，例如在处理纤维化组织时的情况。
