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Imaging Minimal Bacteria at the Nanoscale: a Reliable and Versatile Process to Perform Single-Molecule Localization Microscopy in Mycoplasmas

hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorRIDEAU, Fabien
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorVILLA, Audrey
hal.structure.identifierInterdisciplinary Institute for Neuroscience [Bordeaux] [IINS]
dc.contributor.authorBELZANNE, Pauline
hal.structure.identifierInterdisciplinary Institute for Neuroscience [Bordeaux] [IINS]
dc.contributor.authorVERDIER, Emeline
hal.structure.identifierInterdisciplinary Institute for Neuroscience [Bordeaux] [IINS]
dc.contributor.authorHOSY, Eric
hal.structure.identifierBiologie du fruit et pathologie [BFP]
dc.contributor.authorARFI, Yonathan
dc.date.issued2022
dc.identifier.issn2165-0497
dc.description.abstractEnMycoplasmas are the smallest free-living organisms. These bacteria are important models for both fundamental and synthetic biology, owing to their highly reduced genomes. They are also relevant in the medical and veterinary fields, as they are pathogenic to both humans and most livestock species. Mycoplasma cells have minute sizes, often in the 300- to 800-nm range. As these dimensions are close to the diffraction limit of visible light, fluorescence imaging in mycoplasmas is often poorly informative. Recently developed superresolution imaging techniques can break this diffraction limit, improving the imaging resolution by an order of magnitude and offering a new nanoscale vision of the organization of these bacteria. These techniques have, however, not been applied to mycoplasmas before. Here, we describe an efficient and reliable protocol to perform single-molecule localization microscopy (SMLM) imaging in mycoplasmas. We provide a polyvalent transposon-based system to express the photoconvertible fluorescent protein mEos3.2, enabling photo-activated localization microscopy (PALM) in most Mycoplasma species. We also describe the application of direct stochastic optical reconstruction microscopy (dSTORM). We showcase the potential of these techniques by studying the subcellular localization of two proteins of interest. Our work highlights the benefits of state-of-the-art microscopy techniques for mycoplasmology and provides an incentive to further the development of SMLM strategies to study these organisms in the future..
dc.description.sponsorshipDissection moléculaire du système de clivage d'anticorps des mycoplasmes - ANR-17-CE35-0002
dc.description.sponsorshipExploration de Nouvelles Activités de Manipulation des Anticorps - ANR-21-CE44-0002
dc.language.isoen
dc.publisherAmerican Society for Microbiology
dc.rights.urihttp://creativecommons.org/licenses/by/
dc.subject.enMycoplasma
dc.subject.enPALM
dc.subject.ensingle-molecule localization microscopy
dc.subject.ensuperresolution microscopy
dc.subject.endSTORM
dc.title.enImaging Minimal Bacteria at the Nanoscale: a Reliable and Versatile Process to Perform Single-Molecule Localization Microscopy in Mycoplasmas
dc.typeArticle de revue
dc.identifier.doi10.1128/spectrum.00645-22
dc.subject.halSciences du Vivant [q-bio]
dc.subject.halSciences du Vivant [q-bio]/Médecine humaine et pathologie/Maladies infectieuses
dc.subject.halSciences du Vivant [q-bio]/Biologie animale/Médecine vétérinaire et santé animal
dc.subject.halSciences du Vivant [q-bio]/Médecine humaine et pathologie
bordeaux.journalMicrobiology Spectrum
bordeaux.volumeOnline first
bordeaux.peerReviewedoui
hal.identifierhal-03702545
hal.version1
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-03702545v1
bordeaux.COinSctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=Microbiology%20Spectrum&rft.date=2022&rft.volume=Online%20first&rft.eissn=2165-0497&rft.issn=2165-0497&rft.au=RIDEAU,%20Fabien&VILLA,%20Audrey&BELZANNE,%20Pauline&VERDIER,%20Emeline&HOSY,%20Eric&rft.genre=article


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