A quick flow cytometry protocol to assess Helicobacter pylori viability

doi:10.1016/j.mimet.2020.106043

Journal of Microbiological Methods

Volume 177, October 2020, 106043

https://doi.org/10.1016/j.mimet.2020.106043 Get rights and content

Abstract

Here we present an easy flow cytometry protocol to study the viability of Helicobacter pylori which also enables the detection of even low live bacteria densities. This protocol has potential utility for a fast and accurate assessment of experimental eradication methods against H. pylori.

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Author's contribution

AGM, DGF, JPG and DB had the original concept and idea for the study. DB and TA designed the protocols and the experimental approaches. CA, ACM, AMA, IMG, TS and AJSA performed the experiments. CA and ACM analysed the data and wrote a first draft of the manuscript. All authors reviewed and approved the final version of the manuscript.

Declaration of Competing Interest

None.

Acknowledgements

This work was supported by the Spanish Ministry of Reseach (RyC-2017-21606), Consejería de Educación, Juventud y Deporte de Madrid (Programa de Garantía Juvenil 2015-2016), and Ministerio de Economía y Competitividad (MINECO; projects CTQ2017-84327-P, CTQ2017-83531-R, CTQ2015-71896-REDT, SAF2014-56642-JIN).

References (8)

V. Ambriz-Aviña et al.
Applications of flow cytometry to characterize bacterial physiological responses
Biomed. Res. Int.
(2014)
S. Bury-Moné et al.
Is Helicobacter pylori a true microaerophile?
Helicobacter
(2006)
M. Calvino-Fernández et al.
Helicobacter pylori inactivation and virulence gene damage using a supported sensitiser for photodynamic therapy
Eur. J. Med. Chem.
(2013)
J. Faghri et al.
Morphological and bactericidal effects of different antibiotics on Helicobacter pylori
Jundishapur J. Microbiol.
(2014)

There are more references available in the full text version of this article.

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Citation Excerpt :
The use of rapid and well-established techniques, such as flow cytometry, can be a valuable and effective tool not only to quantify the microbial communities present in the artworks but also to infer their viability and metabolic state using appropriate fluorochromes. Flow cytometry has been successfully used over the years in several fields of research, for the analysis of environmental, pharmaceutical, medical and food samples [17,18,27,28,19–26]. In the context of CH, however, its application is still limited, although some studies have used flow cytometry as a complement to other analytical tools, either in the study of the microorganisms involved in the biodeterioration processes of artworks or the evaluation of the efficiency of potential biocide treatments [29–34].
Flow cytometry was evaluated for the first time for the microbiological characterisation of samples collected from the surfaces of stone and mortar sculptures. The study was carried out on outdoor sculptures located in the district of Porto (Portugal), from which samples were collected using swabs and poly(2-hydroxyethylmethacrylate) (poly(HEMA)) cryogels as non-invasive sampling methods. The use of poly(HEMA) cryogels was tested as an alternative to swabbing due to the difficulties often encountered in sampling stone surfaces and retrieving microorganisms from swab fibres. The quantification of the microorganisms collected by the two sampling methods and the determination of their cell viability were performed using a combination of the fluorochromes thiazole orange (TO) and propidium iodide (PI). Both methods were effective to collect samples for analysis by flow cytometry, which proved to be a technique of interest and potential use for biodeterioration studies of cultural heritage (CH). Data visualisation of viable and non-viable microbial cells was clearer with cryogels, although higher concentrations of total microorganisms were achieved with swabs. Our results show that swab sampling is a more adequate method, although poly(HEMA) cryogels can be an option to consider for specific microbial viability analyses for biodeterioration evaluation of stone artworks.
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¹: Both authors had equal contribution as main authors.

²: Both authors had equal contribution as senior authors.

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NoteA quick flow cytometry protocol to assess Helicobacter pylori viability

Abstract

Section snippets

Author's contribution

Declaration of Competing Interest

Acknowledgements

Applications of flow cytometry to characterize bacterial physiological responses

Biomed. Res. Int.

Is Helicobacter pylori a true microaerophile?

Helicobacter

Helicobacter pylori inactivation and virulence gene damage using a supported sensitiser for photodynamic therapy

Eur. J. Med. Chem.

Morphological and bactericidal effects of different antibiotics on Helicobacter pylori

Jundishapur J. Microbiol.

Note
A quick flow cytometry protocol to assess Helicobacter pylori viability