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Robust single-molecule approach for counting autofluorescent proteins.

hal.structure.identifierCentre de physique moléculaire optique et hertzienne [CPMOH]
dc.contributor.authorCOGNET, Laurent
hal.structure.identifierCentre de physique moléculaire optique et hertzienne [CPMOH]
dc.contributor.authorTARDIN, Catherine
hal.structure.identifierCentre de physique moléculaire optique et hertzienne [CPMOH]
dc.contributor.authorNÉGRIER, Marie-Laure Martin
hal.structure.identifierPhysiologie cellulaire de la synapse [PCS]
dc.contributor.authorBREILLAT, Christelle
hal.structure.identifierPhysiologie cellulaire de la synapse [PCS]
dc.contributor.authorCOUSSEN, Françoise
hal.structure.identifierPhysiologie cellulaire de la synapse [PCS]
dc.contributor.authorCHOQUET, Daniel
hal.structure.identifierCentre de physique moléculaire optique et hertzienne [CPMOH]
dc.contributor.authorLOUNIS, Brahim
dc.date.created2007-07-16
dc.date.issued2008-06-23
dc.identifier.issn1083-3668
dc.description.abstractEnUsing single-molecule microscopy, we present a method to quantify the number of single autofluorescent proteins when they cannot be optically resolved. This method relies on the measurement of the total intensity emitted by each aggregate until it photobleaches. This strategy overcomes the inherent problem of blinking of green fluorescent proteins. In the case of small protein aggregates, our method permits us to describe the mean composition with a precision of one protein. For aggregates containing a large number of proteins, it gives access to the average number of proteins gathered and a signature of the inhomogeneity of the aggregates' population. We applied this methodology to the quantification of small purified citrine multimers.
dc.language.isoen
dc.publisherSociety of Photo-optical Instrumentation Engineers
dc.subject.enaggregation
dc.subject.enbioluminescence
dc.subject.enfluorescence
dc.subject.enmolecular biophysics
dc.subject.enoptical saturable absorption
dc.subject.enproteins
dc.subject.meshAlgorithms
dc.subject.meshGreen Fluorescent Proteins
dc.subject.meshImage Interpretation, Computer-Assisted
dc.subject.meshMicroscopy, Fluorescence
dc.subject.meshMolecular Probe Techniques
dc.subject.meshPhotons
dc.subject.meshSpectrometry, Fluorescence
dc.title.enRobust single-molecule approach for counting autofluorescent proteins.
dc.typeArticle de revue
dc.identifier.doi10.1117/1.2940600
dc.subject.halSciences du Vivant [q-bio]/Biochimie, Biologie Moléculaire/Biophysique
bordeaux.journalJournal of Biomedical Optics
bordeaux.page031216
bordeaux.volume13
bordeaux.issue3
bordeaux.peerReviewedoui
hal.identifierhal-00746847
hal.version1
hal.popularnon
hal.audienceInternationale
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-00746847v1
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