Live-cell imaging and analysis reveal cell phenotypic transition dynamics inherently missing in snapshot data.
Live-cell imaging and analysis reveal cell phenotypic transition dynamics inherently missing in snapshot data.
复制标题
活细胞成像和分析揭示了快照数据中固有缺失的细胞表型转变动态。
DOI：
10.1126/sciadv.aba9319
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发表时间：
2020
期刊：
影响因子：
13.6
通讯作者：
Xing,Jianhua
中科院分区：
文献类型：
--
作者：
Wang,Weikang;Douglas,Diana;Zhang,Jingyu;Kumari,Sangeeta;Enuameh,MetewoSelase;Dai,Yan;Wallace,CallenT;Watkins,SimonC;Shu,Weiguo;Xing,Jianhua
单细胞技术的最新进展催生了一个新兴的领域，即研究细胞如何在一步一步的过程中从一种表型转变为另一种表型。然而，两个重大的技术挑战阻碍了该领域的进一步发展。基于固定细胞的方法可以提供高维表达谱的快照，但在揭示时间信息方面存在基本限制，而基于荧光的活细胞成像方法提供时间信息，但在技术上对多路长期成像具有挑战性。我们首先开发了一个活细胞成像平台，通过结合内源荧光标记和/或高维细胞形态和纹理特征的活细胞成像，将对细胞生理学的扰动降至最低，从而跟踪细胞状态的变化。利用我们的平台和A549 Vim-RFP上皮到间充质转化(EMT)报告细胞系，活细胞轨迹揭示了由于细胞-细胞动态异质性而从快照数据中缺失的平行EMT路径。我们的结果强调了从多路活细胞成像中提取表型转变的动态信息的必要性。
Recent advances in single-cell techniques catalyze an emerging field of studying how cells convert from one phenotype to another, in a step-by-step process. Two grand technical challenges, however, impede further development of the field. Fixed cell–based approaches can provide snapshots of high-dimensional expression profiles but have fundamental limits on revealing temporal information, and fluorescence-based live-cell imaging approaches provide temporal information but are technically challenging for multiplex long-term imaging. We first developed a live-cell imaging platform that tracks cellular status change through combining endogenous fluorescent labeling that minimizes perturbation to cell physiology and/or live-cell imaging of high-dimensional cell morphological and texture features. With our platform and an A549 VIM-RFP epithelial-to-mesenchymal transition (EMT) reporter cell line, live-cell trajectories reveal parallel paths of EMT missing from snapshot data due to cell-cell dynamic heterogeneity. Our results emphasize the necessity of extracting dynamical information of phenotypic transitions from multiplex live-cell imaging.