Rochman Nash D, Yao Kai, Gonzalez Nicolas Perez A, Wirtz Denis, Sun Sean X
Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland 21218, USA.
Department of Mechanical Engineering, Johns Hopkins University, Baltimore, Maryland 21218, USA.
Bio Protoc. 2020 Jun 20;10(12):e3652. doi: 10.21769/BioProtoc.3652.
The measurement of single cell size remains an obstacle towards a deeper understanding of cell growth control, tissue homeostasis, organogenesis, and a wide range of pathologies. Recent advances have placed a spotlight on the importance of cell volume in the regulation of fundamental cell signaling pathways including those known to orchestrate progression through the cell cycle. Here we provide our protocol for the Fluorescence Exclusion Method (FXm); references to the development of FXm; and a brief outlook on future advances in image analysis which may expand the range of problems studied utilizing FXm as well as lower the barrier to entry for groups interested in adding cell volume measurements into their experimental repertoire.
单细胞大小的测量仍然是深入理解细胞生长控制、组织稳态、器官发生以及多种病理状况的一个障碍。最近的进展使人们关注到细胞体积在调节基本细胞信号通路中的重要性，这些信号通路包括那些已知可协调细胞周期进程的通路。在这里，我们提供荧光排除法（FXm）的实验方案；对FXm发展的参考文献；以及对图像分析未来进展的简要展望，这可能会扩大利用FXm研究的问题范围，并降低有兴趣将细胞体积测量纳入其实验方法的研究团队的入门门槛。
