Live cell imaging and analysis reveal cell phenotypic transition dynamics inherently missing in snapshot data

BioRxiv : the Preprint Server for Biology
Weikang WangJianhua Xing

Abstract

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 genome-wide snapshots of cell status 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 EMT reporter line, live cell trajectories reveal parallel paths of epithelial-to-mesenchymal transition missing from snapshot data due to cell-cell heterogeneity. Our results emphasize the necessity of extracting dynamical information of phenotypic transitions from multiplex live cell imaging.

Citations

Apr 9, 2020·Frontiers in Bioengineering and Biotechnology·Priyanka ChakrabortyMohit Kumar Jolly

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Methods Mentioned

BETA
PCA
protein assay
PCR
Extraction
imaging techniques
RNA-seq

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