Rizzoli, Silvio O.

[["dc.bibliographiccitation.journal","Cold Spring Harbor Protocols"],["dc.contributor.author","Hoopmann, P."],["dc.contributor.author","Rizzoli, S. O."],["dc.contributor.author","Betz, W. J."],["dc.date.accessioned","2017-09-07T11:53:37Z"],["dc.date.available","2017-09-07T11:53:37Z"],["dc.date.issued","2012"],["dc.description.abstract","The synaptic vesicle is the essential organelle of the synapse. Many approaches for studying synaptic vesicle recycling have been devised, one of which, the styryl (FM) dye, is well suited for this purpose. FM dyes reversibly stain, but do not permeate, membranes; hence they can specifically label membrane-bound organelles. Their quantum yield is drastically higher when bound to membranes than when in aqueous solution. This protocol describes the imaging of synaptic vesicle recycling by staining and destaining vesicles with FM dyes. Nerve terminals are stimulated (electrically or by depolarization with high K+) in the presence of dye, their vesicles are then allowed to recycle, and finally dye is washed from the chamber. In neuromuscular junction (NMJ) preparations, movements of the muscle must be inhibited if imaging during stimulation is desired (e.g., by application of curare, a potent acetylcholine receptor inhibitor). The main characteristics of FM dyes are also reviewed here, as are recent FM dye monitoring techniques that have been used to investigate the kinetics of synaptic vesicle fusion."],["dc.identifier.doi","10.1101/pdb.prot067603"],["dc.identifier.gro","3145092"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2789"],["dc.language.iso","en"],["dc.notes.intern","Crossref Import"],["dc.notes.status","final"],["dc.relation.issn","1559-6095"],["dc.title","Imaging Synaptic Vesicle Recycling by Staining and Destaining Vesicles with FM Dyes"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dc.type.peerReviewed","no"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","171"],["dc.bibliographiccitation.lastpage","183"],["dc.contributor.author","Hoopmann, P."],["dc.contributor.author","Rizzoli, S. O."],["dc.contributor.author","Betz, W. J."],["dc.contributor.editor","Helmchen, F."],["dc.contributor.editor","Konnerth, A."],["dc.contributor.editor","Yuste, R."],["dc.date.accessioned","2017-09-07T11:45:52Z"],["dc.date.available","2017-09-07T11:45:52Z"],["dc.date.issued","2011"],["dc.identifier.gro","3145550"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/3260"],["dc.language.iso","en"],["dc.notes.intern","lifescience"],["dc.notes.status","final"],["dc.notes.submitter","oschaef1"],["dc.publisher","Cold Spring Harbor Laboratory Press"],["dc.publisher.place","Cold Spring Harbor, New York"],["dc.relation.isbn","978-0-879699-37-6"],["dc.relation.ispartof","Imaging in Neuroscience: A Laboratory Manual"],["dc.title","Imaging Synaptic Vesicle Recycling Using FM Dyes"],["dc.type","book_chapter"],["dc.type.internalPublication","unknown"],["dc.type.peerReviewed","no"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","Cold Spring Harbor Protocols"],["dc.bibliographiccitation.volume","2012"],["dc.contributor.author","Hoopmann, P."],["dc.contributor.author","Rizzoli, S. O."],["dc.contributor.author","Betz, W. J."],["dc.date.accessioned","2017-09-07T11:53:38Z"],["dc.date.available","2017-09-07T11:53:38Z"],["dc.date.issued","2012"],["dc.description.abstract","The synaptic vesicle is the essential organelle of the synapse. Many approaches for studying synaptic vesicle recycling have been devised, one of which, the styryl (FM) dye, is well suited for this purpose. The FM dyes have a unique set of properties that allows them to selectively label recycling vesicles: They reversibly stain, but do not permeate, membranes; hence they can specifically label membrane-bound organelles. Their quantum yield is drastically higher when bound to membranes than when in aqueous solution. FM dyes can also be used as endocytic markers in electron microscopy (EM) through a procedure termed photoconversion (or photooxidation), as described here. Fluorescent dye molecules generate free radicals (reactive oxygen species) when subjected to strong illumination. These short-lived radicals readily oxidize any molecules found in the immediate vicinity of the fluorophore. When photoconversion of FM dyes is performed while the preparation is bathing in diaminobenzidine (DAB), a dark brown precipitate forms after the DAB is oxidized. Thus, illumination turns FM-labeled organelles into dark electron-dense ones. The technique results in a substantial increase in the resolution of FM dye labeling studies (with the obvious caveat that it is restricted to fixed preparations)."],["dc.identifier.doi","10.1101/pdb.prot067611"],["dc.identifier.gro","3145091"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2788"],["dc.language.iso","en"],["dc.notes.intern","Crossref Import"],["dc.notes.status","final"],["dc.relation.issn","1559-6095"],["dc.title","FM Dye Photoconversion for Visualizing Synaptic Vesicles by Electron Microscopy"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dc.type.peerReviewed","no"],["dspace.entity.type","Publication"]]