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Automated, High-Throughput Detection of Bacterial Adherence to Host Cells

Automated, High-Throughput Detection of Bacterial Adherence to Host Cells

Published on: September 17, 2021

Quantification of host-microbe interactions by automated fluorescence microscopy.

Anne-Cécile Haghighat¹, Stéphanie Seveau

¹Departments of Microbiology and Internal Medicine, Division of Infectious Diseases, Center for Microbial Interface Biology, Ohio State University, Columbus, OH, USA.

Journal of Immunological Methods

|November 26, 2009

Summary

We developed an automated microscopy assay to count intracellular pathogens invading host cells. This high-throughput method accurately quantifies pathogen invasion using fluorescent labeling and image analysis.

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Area of Science:

Microbiology
Cell Biology
Biotechnology

Background:

Quantifying intracellular pathogen invasion is crucial for understanding host-pathogen interactions and developing therapeutics.
Existing methods for assessing pathogen invasion can be labor-intensive and lack high-throughput capabilities.

Purpose of the Study:

To develop and validate an automated, high-throughput fluorescence microscopy-based assay for quantifying mammalian cell invasion by intracellular pathogens.

Main Methods:

The assay utilizes automated fluorescence microscopy for image acquisition and processing.
Specific fluorescent labeling distinguishes pathogens, host cells, and cell-associated pathogens.
Direct counting of labeled entities enables quantification of invasion.

Main Results:

The assay accurately quantifies pathogen invasion by directly counting labeled components.
Automated image analysis facilitates high-throughput data generation.
The method was successfully validated using Listeria monocytogenes as a model pathogen.

Conclusions:

This automated fluorescence microscopy assay provides a robust and efficient tool for high-throughput quantification of intracellular pathogen invasion.
The assay is adaptable for studying various intracellular pathogens and host-pathogen systems.