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dc.contributor.authorStröhl, Florian
dc.contributor.authorLin, Julie Qiaojin
dc.contributor.authorTartwijk, Francesca W. van
dc.contributor.authorWong, Hovy Ho Wai
dc.contributor.authorHolt, Christine E.
dc.contributor.authorKaminski, Clemens F.
dc.date.accessioned2023-09-26T12:46:04Z
dc.date.available2023-09-26T12:46:04Z
dc.date.issued2020-04-29
dc.description.abstractSingle-molecule translation imaging (SMTI) is a straightforward technique for the direct quantification of local protein synthesis. The protein of interest is fused to a fast-folding and fast-bleaching fluorescent protein, allowing one to monitor the appearance of individual fluorescence events after photobleaching of pre-existing proteins in the cell under investigation. The translation of individual molecules is then indicated by photon bursts of sub-second length that appear over a dark background. The method thus shares attributes with fluorescence recovery after photobleaching (FRAP) microscopy. Resulting datasets are similar to those generated by localization-based super-resolution microscopy techniques and can be used both to generate density maps of local protein production and to quantify the kinetics of local synthesis. The detailed protocol described in this chapter uses a Venus-β-actin fusion construct to visualize and measure the β-actin mRNA translational activity in Xenopus retinal ganglion cell growth cones upon Netrin-1 stimulation, which can be readily adapted for detecting translation events of other mRNAs in various cell types.en_US
dc.identifier.citationStröhl F, Lin JQ, Tartwijk FWv, Wong HHW, Holt CE, Kaminski CF: A Protocol for Single-Molecule Translation Imaging in Xenopus Retinal Ganglion Cells. In: Yamamoto N, Okada Y. Single Molecule Microscopy in Neurobiology, 2020. Springer p. 295-308en_US
dc.identifier.cristinIDFRIDAID 1809094
dc.identifier.doihttps://doi.org/10.1007/978-1-0716-0532-5_14
dc.identifier.isbn978-1-0716-0532-5
dc.identifier.issn0893-2336
dc.identifier.issn1940-6045
dc.identifier.urihttps://hdl.handle.net/10037/31214
dc.language.isoengen_US
dc.publisherSpringer Natureen_US
dc.relation.ispartofseriesNM,volume 154en_US
dc.rights.accessRightsopenAccessen_US
dc.rights.holderCopyright 2020 The Author(s)en_US
dc.titleA Protocol for Single-Molecule Translation Imaging in Xenopus Retinal Ganglion Cellsen_US
dc.type.versionsubmittedVersionen_US
dc.typeChapteren_US
dc.typeBokkapittelen_US


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