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Recent Advances on Multi-Parameter Flow Cytometry to Characterize Antimicrobial Treatments
Lucie Léonard1, Lynda Bouarab Chibane1, Balkis Ouled Bouhedda1
1Univ Lyon, Université Claude Bernard Lyon 1, ISARA Lyon, BioDyMIA (Bioingénierie et Dynamique Microbienne aux Interfaces Alimentaires), Equipe Mixte d'Accueil n°3733, IUT Lyon 1 Bourg en Bresse, France.
Abstract:
The investigation on antimicrobial mechanisms is a challenging and crucial issue in the fields of food or clinical microbiology, as it constitutes a prerequisite to the development of new antimicrobial processes or compounds, as well as to anticipate phenomenon of microbial resistance. Nowadays it is accepted that a cells population exposed to a stress can cause the appearance of different cell populations and in particular sub-lethally compromised cells which could be defined as viable but non-culturable (VBNC). Recent advances on flow cytometry (FCM) and especially on multi-parameter flow cytometry (MP-FCM) provide the opportunity to obtain high-speed information at real time on damage at single-cell level. This review gathers MP-FCM methodologies based on individual and simultaneous staining of microbial cells employed to investigate their physiological state following different physical and chemical antimicrobial treatments. Special attention will be paid to recent studies exploiting the possibility to corroborate MP-FCM results with additional techniques (plate counting, microscopy, spectroscopy, molecular biology techniques, membrane modeling) in order to elucidate the antimicrobial mechanism of action of a given antimicrobial treatment or compound. The combination of MP-FCM methodologies with these additional methods is namely a promising and increasingly used approach to give further insight in differences in microbial sub-population evolutions in response to antimicrobial treatments.
Insights
Multi-parameter flow cytometry (MP-FCM) reveals microbial responses to antimicrobial treatments. This technique identifies viable but non-culturable cells and elucidates antimicrobial mechanisms by combining multiple data types.
Area of Science:
- Microbiology
- Cell Biology
- Analytical Chemistry
Background:
- Understanding antimicrobial mechanisms is vital for developing new treatments and combating microbial resistance.
- Microbial populations under stress can develop distinct subpopulations, including viable but non-culturable (VBNC) cells.
- Single-cell analysis is crucial for understanding heterogeneous microbial responses.
Purpose of the Study:
- To review multi-parameter flow cytometry (MP-FCM) methodologies for assessing microbial physiological states after antimicrobial treatments.
- To highlight the integration of MP-FCM with other techniques for a comprehensive understanding of antimicrobial action.
- To provide insights into microbial sub-population dynamics under antimicrobial stress.
Main Methods:
- Review of MP-FCM techniques using individual and simultaneous cell staining.
- Analysis of microbial physiological states following physical and chemical antimicrobial treatments.
- Integration of MP-FCM data with complementary methods like plate counting, microscopy, spectroscopy, and molecular biology.
Main Results:
- MP-FCM enables high-speed, real-time, single-cell analysis of microbial damage.
- The technique effectively distinguishes between different microbial subpopulations, including VBNC cells.
- Combining MP-FCM with other methods provides deeper insights into antimicrobial mechanisms of action.
Conclusions:
- MP-FCM is a powerful tool for investigating antimicrobial mechanisms and microbial responses.
- Integrating MP-FCM with complementary techniques offers a robust approach to studying microbial physiology.
- This integrated approach is essential for advancing antimicrobial research and developing novel strategies.
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