化学
细胞生物学
生物物理学
显微镜
细胞骨架
膜蛋白
荧光显微镜
膜
细胞膜
活体细胞成像
生物
作者
Charlène Cornilleau,Arnaud Chastanet,Cyrille Billaudeau,Rut Carballido-López
出处
期刊:Methods of Molecular Biology
日期:2020-01-01
卷期号:2101: 123-133
被引量:3
标识
DOI:10.1007/978-1-0716-0219-5_8
摘要
MreB proteins are actin homologs present in nonspherical bacteria. They assemble into membrane-associated discrete filamentous structures that exhibit different dynamic behaviors along the bacterial sidewalls. Total internal reflection fluorescence (TIRF) microscopy, a sensitive method for studying molecular events at cell surfaces with high contrast and temporal resolution, is a method of choice to characterize the localization and dynamics of cortical MreB assemblies in vivo. This chapter describes the methods for visualizing fluorescently tagged MreB proteins in live Bacillus subtilis cells. We detail how to (1) grow B. subtilis strains for reproducible TIRF observations, (2) immobilize cells on agarose pads and (3) in CellASIC® microfluidic plates, and (4) acquire TIRF images and time lapses.
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