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Correlative nano-imaging of metals and proteins in primary neurons by synchrotron X-ray fluorescence and STED super resolution microscopy: Experimental validation

hal.structure.identifierLaboratoire de Physique des Deux Infinis Bordeaux [LP2I - Bordeaux]
dc.contributor.authorORTEGA, Richard
hal.structure.identifierLaboratoire de Physique des Deux Infinis Bordeaux [LP2I - Bordeaux]
dc.contributor.authorROUDEAU, Stéphane
hal.structure.identifierLaboratoire de Physique des Deux Infinis Bordeaux [LP2I - Bordeaux]
dc.contributor.authorCARMONA, Asuncion
dc.date.accessioned2022-09-15T12:34:44Z
dc.date.available2022-09-15T12:34:44Z
dc.date.issued2022
dc.identifier.urihttps://oskar-bordeaux.fr/handle/20.500.12278/145321
dc.description.abstractEnBackgroundIt is becoming increasingly clear that biological metals such as iron, copper or zinc are involved in synaptic functions, and in particular in the mechanisms of synaptogenesis and subsequent plasticity. Understanding the role of metals on synaptic functions is a difficult challenge due to the very low concentration of these elements in neurons and to the submicrometer size of synaptic compartments.New methodTo address this challenge we have developed a correlative nano-imaging approach combining metal and protein detection. First, stimulated emission depletion (STED) microscopy, a super resolution optical microscopy technique, is applied to locate fluorescently labeled proteins. Then, synchrotron radiation induced X-ray fluorescence (SXRF) is performed on the same regions of interest, e.g. synaptic compartments.ResultsWe present here the principle scheme that allows this correlative nano-imaging and its experimental validation. We applied this correlative nano-imaging to the study of the physiological distribution of metals in synaptic compartments of primary rat hippocampal neurons. We thus compared the nanometric distribution of metals with that of synaptic proteins, such as PSD95 or cytoskeleton proteins.Comparison with existing method(s)Compared to correlative imaging approaches currently used to characterize synaptic structures, such as electron microscopy correlated with optical fluorescence, our approach allows for ultra-sensitive detection of trace metals using highly focused synchrotron radiation beams.ConclusionWe provide proof-of-principle for correlative imaging of metals and proteins at the synaptic scale and discuss the present limitations and future developments in this area
dc.language.isoen
dc.title.enCorrelative nano-imaging of metals and proteins in primary neurons by synchrotron X-ray fluorescence and STED super resolution microscopy: Experimental validation
dc.typeArticle de revue
dc.identifier.doi10.1016/j.jneumeth.2022.109702
dc.subject.halPhysique [physics]/Physique [physics]/Instrumentations et Détecteurs [physics.ins-det]
bordeaux.journalJ.Neurosci.Methods
bordeaux.page109702
bordeaux.volume381
bordeaux.hal.laboratoriesLaboratoire de Physique des Deux Infinis de Bordeaux (LP2I) - UMR 5797*
bordeaux.institutionUniversité de Bordeaux
bordeaux.institutionCNRS
bordeaux.peerReviewedoui
hal.identifierhal-03771841
hal.version1
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-03771841v1
bordeaux.COinSctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=J.Neurosci.Methods&rft.date=2022&rft.volume=381&rft.spage=109702&rft.epage=109702&rft.au=ORTEGA,%20Richard&ROUDEAU,%20St%C3%A9phane&CARMONA,%20Asuncion&rft.genre=article


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