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Related Experiment Video

Updated: Mar 31, 2026

Analysis of Side Population in Solid Tumor Cell Lines

Analysis of Side Population in Solid Tumor Cell Lines

Published on: February 23, 2021

Analyzing Persister Physiology with Fluorescence-Activated Cell Sorting.

Mehmet A Orman¹, Theresa C Henry^2,3, Christina J DeCoste²

¹Department of Chemical and Biological Engineering, Princeton University, 205 Hoyt Laboratory, Princeton, NJ, 08544, USA.

Methods in Molecular Biology (Clifton, N.J.)

|October 16, 2015

Summary

This summary is machine-generated.

Bacterial persister cells, crucial for antibiotic tolerance and infection relapse, can now be studied using novel fluorescence-activated cell sorting (FACS) protocols to reveal their metabolic and growth states.

Keywords:

Antibiotic Fluorescence-activated cell sorting (FACS)Persister Phenotypic heterogeneity Redox sensor green (RSG)Viable but non-culturable cell (VBNC)

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Isolation of Normal and Cancer-associated Fibroblasts from Fresh Tissues by Fluorescence Activated Cell Sorting FACS

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Published on: January 14, 2013

Area of Science:

Microbiology
Cell Biology
Antibiotic Resistance

Background:

Bacterial persisters are phenotypic variants with high antibiotic tolerance, implicated in recurrent infections.
Limited knowledge of persister physiology hinders development of treatments for recalcitrant infections.
Isolation challenges due to low abundance and similarity to viable but non-culturable cells (VBNCs) impede persister research.

Purpose of the Study:

To describe detailed fluorescence-activated cell sorting (FACS) protocols for analyzing bacterial persister physiology.
To enable quantification and characterization of persister cells and their heterogeneity.
To overcome technical hurdles in persister isolation and phenotypic analysis.

Main Methods:

Utilizing fluorescence-activated cell sorting (FACS) to isolate and quantify bacterial persister cells.
Developing a protocol to assay the metabolic state of persisters using fluorescent metabolic stains.
Developing a protocol to assay the growth state of persisters using fluorescent proteins.

Main Results:

Established FACS-based assays provide persister phenotype distributions.
These distributions allow comparison with the entire bacterial population.
The protocols facilitate examination of persister heterogeneity and physiological states.

Conclusions:

FACS protocols offer a robust method for analyzing persister cell physiology.
These methods advance the understanding of persister cells' metabolic and growth states.
This research paves the way for novel therapeutic strategies against antibiotic-tolerant bacteria.