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Live-cell imaging and analysis reveal cell phenotypic transition dynamics inherently missing in snapshot data
Weikang Wang1, Diana Douglas2, Jingyu Zhang1
1Department of Computational and Systems Biology, University of Pittsburgh, Pittsburgh, PA 15232, USA.
Science Advances
|September 12, 2020
Summary
New live-cell imaging tracks cell changes, revealing parallel paths in epithelial-to-mesenchymal transition (EMT) missed by static methods. This dynamic approach is crucial for understanding cell phenotype transitions.
Area of Science:
- Cell biology
- Biophysics
- Single-cell analysis
Background:
- Single-cell techniques are advancing the study of cellular phenotype transitions.
- Current methods face challenges: fixed-cell approaches lack temporal data, while live-cell imaging struggles with long-term multiplexing.
Purpose of the Study:
- To develop a novel live-cell imaging platform for tracking cellular status changes.
- To overcome limitations of existing methods in capturing dynamic phenotypic transitions.
Main Methods:
- Developed a live-cell imaging platform using endogenous fluorescent labeling and high-dimensional morphological/texture feature analysis.
- Utilized an A549 VIM-RFP epithelial-to-mesenchymal transition (EMT) reporter cell line.
- Tracked live-cell trajectories to capture dynamic cellular events.
Main Results:
- The platform successfully tracked cellular status changes, revealing parallel EMT pathways.
- Live-cell trajectories uncovered dynamic cell-cell heterogeneity previously missed by snapshot data.
- Demonstrated the platform's capability for multiplex, long-term imaging.
Conclusions:
- Dynamic information extraction from multiplex live-cell imaging is essential for studying phenotypic transitions.
- The developed platform offers a powerful tool for investigating complex cellular dynamics.
- Highlights the importance of live-cell imaging for a comprehensive understanding of cell fate decisions.
