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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.issued2022-02-02
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 of both humans and most livestock species. Mycoplasma cells have minute sizes, often in the 300-800 nanometers range. As these dimensions are close to the diffraction limit of visible light, fluorescence imaging in mycoplasmas is often poorly informative. Recently developed Super-Resolution 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 photo-convertible 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 SMLM strategies to study these organisms in the future.
dc.language.isoen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/
dc.subjectmollicute
dc.subjectmycoplasme
dc.subjectbactérie pathogène
dc.subjectmicroscopie
dc.subjectmaladie infectieuse
dc.subjectmycoplasma
dc.title.enImaging minimal bacteria at the nanoscale: a reliable and versatile process to perform Single Molecule Localization Microscopy in mycoplasmas
dc.typeDocument de travail - Pré-publication
dc.identifier.doi10.1101/2022.02.02.478797
dc.subject.halSciences du Vivant [q-bio]
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
hal.identifierhal-03555154
hal.version1
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-03555154v1
bordeaux.COinSctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.date=2022-02-02&rft.au=RIDEAU,%20Fabien&VILLA,%20Audrey&BELZANNE,%20Pauline&VERDIER,%20Emeline&HOSY,%20Eric&rft.genre=preprint


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