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- W2012311273 abstract "We combine single molecule photobleaching with super-resolution fluorescence imaging to count the absolute number of proteins in small oligomers using PALM.First we employed single molecule imaging and stepwise photobleaching of fluorescently tagged glycine-gated ion channels (GlyR), to determine subunit stoichiometry of this neurotransmitter to be 3α1:2β (Durisic et. al JNeuroSci 2012). Then the channel was used as a template to precisely characterize photoactivation and photoconversion efficiency at the single molecule level of most commonly used fluorescent proteins in PALM microscopy. using stepwise photobleaching we characterized the effects of dense sample labeling and determined that only a subset of fluorescent proteins can be efficiently photoacivated or photoconverted. We could then directly relate these results to the number of GlyR subunits per channel counted by means of PALM microscopy. Our findings have important implications for super-resolution microscopy, particularly in the context of quantitative imaging and co-localization in multi-color super-resolution experiments." @default.
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- W2012311273 date "2013-01-01" @default.
- W2012311273 modified "2023-10-07" @default.
- W2012311273 title "Effects of Photoactivation Efficiency on Single Molecule Counting in PALM" @default.
- W2012311273 doi "https://doi.org/10.1016/j.bpj.2012.11.2903" @default.
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