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Estimating microbial growth is essential for understanding population dynamics and environmental adaptations. Indirect methods provide valuable insights by measuring parameters such as turbidity, metabolic activity, and biomass, enabling efficient and reproducible assessments.During exponential growth, microbial cells scatter light proportionally to their biomass, a principle used in turbidity measurements. About one million cells per milliliter produce detectable scattering, which a...
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Quantification of Microbial Fluorescent Sensors During Live Intracellular Infections.

Erez Mills1, Erik Petersen2

  • 1Department of Animal Sciences, Robert H. Smith Faculty of Agriculture, Food, and Environment, The Hebrew University of Jerusalem, Rehovot, Israel. erez.mills@mail.huji.ac.il.

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Summary

This study introduces a novel fluorescent assay for quantifying pathogen behavior within host cells. This method enables single-cell analysis, revealing pathogen subpopulations missed by traditional population-level assays.

Keywords:
Cyclic-di-GMPFluorescent reporterIntracellular pathogenLive cell microscopySalmonellaSingle-cell quantification

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Area of Science:

  • Microbiology
  • Cell Biology
  • Infectious Diseases

Background:

  • Pathogen-host interactions during intracellular growth are crucial for disease pathogenesis.
  • Quantifying pathogen responses inside host cells is challenging due to low pathogen biomolecule concentrations.
  • Fluorescent proteins are increasingly used as reporters and biosensors for studying pathogen behavior.

Purpose of the Study:

  • To develop a sensitive fluorescent assay for quantifying pathogen intracellular behavior.
  • To enable single-cell level analysis of pathogen responses during infection.
  • To overcome limitations of population-level assays in studying pathogen dynamics.

Main Methods:

  • Development of a novel fluorescent assay for pathogen intracellular growth.
  • Utilizing fluorescent proteins as transcriptional reporters and biosensors.
  • Implementing single-cell level quantification of pathogen responses.

Main Results:

  • The fluorescent assay allows for sensitive, live observation of pathogen responses.
  • Single-cell analysis revealed pathogen subpopulations previously undetected.
  • The method overcomes challenges associated with low pathogen biomolecule concentrations.

Conclusions:

  • This fluorescent assay provides a powerful tool for studying pathogen-host interactions at the single-cell level.
  • The technique enhances our understanding of pathogen dynamics within host cells.
  • It offers a sensitive method for quantifying pathogen behavior and uncovering hidden subpopulations.