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Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Updated: Jun 6, 2025

Monitoring Spatial Segregation in Surface Colonizing Microbial Populations

Monitoring Spatial Segregation in Surface Colonizing Microbial Populations

Published on: October 29, 2016

Biofilm dispersal patterns revealed using far-red fluorogenic probes.

Jojo A Prentice¹, Sandhya Kasivisweswaran¹, Robert van de Weerd^1,2,3

¹Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania, United States of America.

|November 25, 2024

Summary

This summary is machine-generated.

Researchers developed a new imaging method to study how bacteria disperse from biofilms. This technique revealed new details about bacterial dispersal patterns and identified cells that do not disperse.

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Last Updated: Jun 6, 2025

Monitoring Spatial Segregation in Surface Colonizing Microbial Populations

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Published on: October 29, 2016

Concurrent Quantification of Cellular and Extracellular Components of Biofilms

Concurrent Quantification of Cellular and Extracellular Components of Biofilms

Published on: December 10, 2013

Methods for Characterizing the Co-development of Biofilm and Habitat Heterogeneity

Methods for Characterizing the Co-development of Biofilm and Habitat Heterogeneity

Published on: March 11, 2015

Area of Science:

Microbiology
Cell Biology
Biophysics

Background:

Bacteria form biofilms, complex communities crucial for colonization and infection.
Dispersal from biofilms is vital for bacterial spread but poorly understood at the single-cell level.
Limitations of traditional imaging methods hinder the study of dispersal in dense, oxygen-deprived biofilms.

Purpose of the Study:

To develop and apply a novel cell-labeling strategy for high-resolution, long-term imaging of bacterial biofilm dispersal.
To investigate the single-cell mechanisms and spatial dynamics of biofilm dispersal in *Vibrio cholerae*.
To identify novel micro-scale patterns and cellular behaviors during the dispersal process.

Main Methods:

Developed a cell-labeling strategy using fluorogen-activating proteins (FAPs) and far-red dyes for functional imaging in biofilms.
Utilized long-term imaging to characterize dispersal dynamics at unprecedented resolution in *Vibrio cholerae* biofilms.
Analyzed dispersal patterns, including cell departure, biofilm compression, and regional cell motion heterogeneity.

Main Results:

Dispersal initiates at the biofilm periphery, with approximately 25% of cells exhibiting no dispersal.
Novel micro-scale patterns observed during dispersal include biofilm compression and heterogeneous cell motion.
Mutants with altered dispersal rates showed attenuated or homogenized dispersal patterns, impacting local mechanical properties.

Conclusions:

Fluorogen-activating proteins (FAPs) provide a powerful tool for high-resolution microbial dynamics studies in complex environments.
Characterized fundamental insights into the mechanisms of bacterial biofilm dispersal and pathogen dissemination.
Revealed previously unobserved single-cell behaviors and spatial patterns governing biofilm dispersal.