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Live-Cell Fluorescence Microscopy to Investigate Subcellular Protein Localization and Cell Morphology Changes in Bacteria
Published on: November 23, 2019
High-throughput, subpixel precision analysis of bacterial morphogenesis and intracellular spatio-temporal dynamics
Oleksii Sliusarenko1, Jennifer Heinritz, Thierry Emonet
1Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06520, USA.
Molecular Microbiology
|March 19, 2011
Summary
We developed MicrobeTracker, an image analysis tool for bacteria, to study cell shape and internal organization. This tool revealed how protein concentration affects the Escherichia coli Min oscillator and uncovered new insights into Caulobacter crescentus cell division.
Area of Science:
- Microbiology
- Cell Biology
- Biophysics
Background:
- Bacteria exhibit diverse shapes and complex intracellular organization crucial for cellular processes.
- Studying spatio-temporal dynamics requires automated analysis of large microscopy datasets.
Purpose of the Study:
- To develop a high-throughput image analysis program, MicrobeTracker, for precise bacterial cell segmentation and lineage tracking.
- To investigate the impact of protein concentration on the Escherichia coli Min oscillator.
- To explore morphogenesis and cell cycle regulation in Caulobacter crescentus.
Main Methods:
- Development of MicrobeTracker for subpixel cell outlining and segmentation, including crowded images and mini-colonies.
- Integration of SpotFinder for tracking fluorescently labeled molecules within cells.
- Application of MicrobeTracker to analyze Escherichia coli and Caulobacter crescentus microscopy data.
Main Results:
- MicrobeTracker enables accurate cell lineage tracking and analysis of intracellular component dynamics.
- Escherichia coli Min oscillator dynamics are dependent on Min protein concentration, revealing robustness limitations.
- The fraction of oscillating MinD proteins increases with cell length in E. coli.
- Position-dependent silencing of chromosomal origins during Caulobacter crescentus cell division was observed and linked to regulation by division.
Conclusions:
- MicrobeTracker is a versatile tool for quantitative analysis of bacterial cell dynamics.
- Bacterial oscillators and cell division processes exhibit complex regulation influenced by protein concentration, cell length, and spatial organization.
