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A fluorescence microscopy based genetic screen to identify mutants altered for interactions with host cells
R L Guy1, L A Gonias, M A Stein
1Department of Microbiology, University of Vermont, Burlington, VT 05405-0068, USA.
Journal of Microbiological Methods
|October 6, 2000
Summary
This study introduces Special Optics (SO) plates for high-throughput screening of bacterial mutants. This method enables efficient identification of microbial virulence factors using fluorescence microscopy.
Area of Science:
- Microbiology
- Cell Biology
- Microbial Pathogenesis
Background:
- Immunofluorescence microscopy is crucial for studying microbial-host interactions.
- High-throughput screening of bacterial mutants is essential for understanding pathogenesis.
- Current methods are limited by the lack of suitable materials for large-scale analysis.
Purpose of the Study:
- To evaluate 96-well Special Optics (SO) plates for high-throughput screening of bacterial mutants.
- To identify Salmonella typhimurium mutants defective in disrupting host cell endocytic compartments.
- To facilitate the characterization of microbial phenotypes using fluorescence microscopy.
Main Methods:
- Utilized 96-well Special Optics (SO) plates for microbial screening.
- Examined 11,520 Salmonella typhimurium MudJ mutants.
- Assessed the loss of ability to disrupt host cell endocytic compartments via fluorescence phenotype.
Main Results:
- Successfully screened a large library of Salmonella typhimurium mutants using SO plates.
- Identified mutants with altered interactions with host cell endocytic compartments.
- Characterized the formation of lysosomal membrane glycoprotein (lgp) containing tubules during Salmonella infection.
Conclusions:
- Special Optics (SO) plates are effective for high-throughput screening of microbial phenotypes.
- This approach enables direct screening for fluorescence-based virulence phenotypes.
- Facilitates the study of microbial pathogenesis and host-pathogen interactions.