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Bacterial and archaeal cells exhibit remarkable diversity in shape and structure, critical in their adaptability and functionality. Among bacteria, the most commonly observed shapes include cocci and bacilli. Cocci are spherical and may exist singly or in groupings such as pairs (diplococci), chains (streptococci), clusters (staphylococci), or tetrads. Bacilli, in contrast, are rod-shaped and can also occur as single cells, in pairs, or chains, depending on their environmental and genetic...
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Author Spotlight: Advancing Research in Microbial Autoaggregation Using Imaging Flow Cytometry
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Imaging Flow Cytometry Demonstrates Physiological and Morphological Diversity within Treated Probiotic Bacteria

Jakub Kiepś1, Wojciech Juzwa1, Radosław Dembczyński1

  • 1Department of Biotechnology and Food Microbiology, Poznan University of Life Sciences, Wojska Polskiego 48, 60-627 Poznań, Poland.

International Journal of Molecular Sciences
|April 13, 2023
PubMed
Summary

Imaging flow cytometry (IFC) accurately enumerates probiotic bacteria, offering insights into cell physiology and morphology. This method identifies viable but nonculturable probiotic cells by analyzing metabolic activity and membrane integrity.

Keywords:
fluid bed dryinglactic acid bacteriaquality controlrapid assessmentstress factorsviability

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Area of Science:

  • Microbiology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Probiotic bacteria require robust enumeration methods for quality control.
  • Conventional methods like plate counts and traditional flow cytometry have limitations in assessing bacterial viability and physiology.
  • Introducing stress during culturing is an alternative to protectants for probiotic stability.

Purpose of the Study:

  • To evaluate imaging flow cytometry (IFC) as a method for accurate probiotic bacterial enumeration.
  • To compare IFC with traditional methods for assessing bacterial viability and physiological state.
  • To identify viable but nonculturable (VBNC) probiotic cells using IFC.

Main Methods:

  • Probiotic bacteria were subjected to stresses during culturing.
  • Imaging flow cytometry (IFC) was employed for bacterial enumeration and analysis.
  • Complementary dyes (RedoxSensorTM Green, propidium iodide) were used to assess metabolic activity and membrane integrity.
  • Cell sorting was performed to analyze sorted populations on different growth media (MRS-Agar, MRS broth).

Main Results:

  • IFC provides detailed insights into bacterial physiology and morphology beyond traditional flow cytometry.
  • Distinct bacterial populations were identified based on metabolic activity and membrane integrity.
  • Cells exhibiting intermediate metabolic activity and membrane damage were characterized as viable but nonculturable (VBNC).

Conclusions:

  • IFC is a powerful tool for accurate probiotic enumeration and detailed physiological assessment.
  • IFC enables the identification and characterization of VBNC probiotic populations.
  • This method offers an advantage over conventional techniques for quality control of probiotic formulations.