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- W2893497029 abstract "Quantitative single molecule localization microscopy (qSMLM) is a powerful approach to study in situ protein organization. However, uncertainty regarding the photophysical properties of fluorescent reporters can bias the interpretation of detected localizations and subsequent quantification. Furthermore, strategies to efficiently detect endogenous proteins are often constrained by label heterogeneity and reporter size. Here, a new surface assay for molecular isolation (SAMI) was developed for qSMLM and used to characterize photophysical properties of fluorescent proteins and dyes. SAMI-qSMLM afforded robust quantification. To efficiently detect endogenous proteins, we used fluorescent ligands that bind to a specific site on engineered antibody fragments. Both the density and nano-organization of membrane-bound epidermal growth factor receptors (EGFR, HER2, and HER3) were determined by a combination of SAMI, antibody engineering, and pair-correlation analysis. In breast cancer cell lines, we detected distinct differences in receptor density and nano-organization upon treatment with therapeutic agents. This new platform can improve molecular quantification and can be developed to study the local protein environment of intact cells." @default.
- W2893497029 created "2018-10-05" @default.
- W2893497029 creator A5006016631 @default.
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- W2893497029 date "2018-09-26" @default.
- W2893497029 modified "2023-10-16" @default.
- W2893497029 title "A Platform To Enhance Quantitative Single Molecule Localization Microscopy" @default.
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- W2893497029 doi "https://doi.org/10.1021/jacs.8b04939" @default.
- W2893497029 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/6187371" @default.
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- W2893497029 hasPublicationYear "2018" @default.
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