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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

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The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Analyzing Cellular Internalization of Nanoparticles and Bacteria by Multi-spectral Imaging Flow Cytometry
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Imaging flow cytometry analysis of intracellular pathogens.

Viraga Haridas1, Shahin Ranjbar1, Ivan A Vorobjev2

  • 1Program in Cellular and Molecular Medicine, Boston Children's Hospital, Harvard Medical School, United States; Department of Pediatrics, Harvard Medical School, United States.

Methods (San Diego, Calif.)
|September 20, 2016
PubMed
Summary

Imaging flow cytometry offers a powerful method to study host-pathogen interactions, particularly for intracellular pathogens. This technique quantitatively analyzes cellular events, advancing our understanding of infection biology.

Keywords:
Cellular heterogeneityColocalizationFeature FinderFluorescent proteinImaging flow cytometryIntracellular pathogenMycobacteria tuberculosisPhagosome maturationRab5Rab7Toxoplasma gondii

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Area of Science:

  • Infection Biology
  • Cellular Microbiology
  • Immunology

Background:

  • Intracellular pathogens pose significant challenges in infection biology.
  • Understanding host-pathogen interactions is crucial for developing effective treatments.
  • Traditional methods often lack the quantitative resolution needed for complex cellular interactions.

Purpose of the Study:

  • To provide an overview of imaging flow cytometry applications in studying host-pathogen interactions.
  • To highlight the quantitative analysis capabilities for intracellular infections.
  • To demonstrate the utility of imaging flow cytometry with specific pathogen examples.

Main Methods:

  • Utilizing imaging flow cytometry with specialized analytic software (e.g., IDEAS™ and Feature Finder).
  • Analyzing hundreds of quantified features for cellular and subcellular events.
  • Employing fluorescently labeled prokaryotic and eukaryotic pathogens (e.g., *Toxoplasma gondii*, *Mycobacterium tuberculosis*) in human cell lines.

Main Results:

  • Imaging flow cytometry enables quantitative analysis of host cell-pathogen interactions.
  • Key metrics include cell counting, internalization scores, and subcellular co-localization patterns.
  • The Feature Finder algorithm identifies statistically significant differences in image galleries.

Conclusions:

  • Imaging flow cytometry provides a powerful approach to quantitatively study host-pathogen interactions.
  • This methodology enhances the understanding of host control mechanisms against intracellular pathogens.
  • The combination of imaging flow cytometry and advanced software offers new insights into infection dynamics.