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• W3035544103 abstract "Resolution is a key feature in microscopy which allows the visualization of the fine structure of cells. Much of the life processes within these cells depend on the three-dimensional (3D) complexity of these structures. Optical super-resolution microscopies are currently the preferred choice of molecular and cell biologists who seek to visualize the organization of specific protein species at the nanometer scale. Traditional super-resolution microscopy techniques have often been limited by sample thickness, axial resolution, specialist optical instrumentation and computationally-demanding software for assembling the images. In this chapter we detail the protocol, “enhanced expansion microscopy” (EExM), which combines X10 expansion microscopy with Airyscan confocal microscopy. EExM enables 15 nm lateral (and 35 nm axial) resolution, and is a relatively cheap, accessible option allowing single protein resolution for the non-specialist optical microscopists. We illustrate how EExM has been utilized for mapping the 3D topology of intracellular protein arrays at sample depths which are not always compatible with some of the traditional super-resolution techniques. We demonstrate that antibody markers can recognize and map post-translational modifications of individual proteins in addition to their 3D positions. Finally, we discuss the current uncertainties and validations in EExM which include the isotropy in gel expansion and assessment of the expansion factor (of resolution improvement)." @default.
• W3035544103 created "2020-06-19" @default.
• W3035544103 creator A5014123456 @default.
• W3035544103 creator A5068412492 @default.
• W3035544103 date "2021-01-01" @default.
• W3035544103 modified "2023-09-26" @default.
• W3035544103 title "Enhanced expansion microscopy to measure nanoscale structural and biochemical remodeling in single cells" @default.
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• W3035544103 doi "https://doi.org/10.1016/bs.mcb.2020.04.019" @default.
• W3035544103 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/33478687" @default.
• W3035544103 hasPublicationYear "2021" @default.
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