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Resolution Matters : Correlating Quantitative Proteomics and Nanoscale-Precision Microscopy for Reconstructing Synapse Identity
Verfasser / VerfasserinHarkany, Tibor ; Lubec, Gert ; Miklosi, Andras Gabor ; Del Favero, Gioriga ; Marko, Doris
Erschienen in
Proteomics, 2018, Jg. 18, H. 14, S. 1800139
ErschienenWiley-Blackwell, 2018
DokumenttypAufsatz in einer Zeitschrift
Schlagwörter (EN)dSTORM / immunohistochemistry / spatial proteomics / synapse
URNurn:nbn:at:at-ubmuw:3-1021 Persistent Identifier (URN)
 Das Werk ist frei verfügbar
Resolution Matters [0.39 mb]
Zusammenfassung (Englisch)

For more than a century, the precision at which any protein (or RNA) could be localized in living cells depends on the spatial resolution of microscopy. Light microscopy, even recently benchmarked laserscanning microscopy, is inherently liable to the diffraction limit of visible light. Electron microscopy that had existed as the only alternative for decades is, in turn, of low throughput and sensitive to processing artefacts. Therefore, researchers have looked for alternative technologies particularly with evergrowing interest in resolving structural underpinnings of cellular heterogeneity in the human body. Computational (“in silico”) predictions provided only partial solutions given the incompleteness of existing databases and erroneous assumptions on evolutionarily conserved sequence homology across species. A breakthrough that facilitates subcellular protein localization came with the introduction of “superresolution” microscopy, which yields 2060 nm resolution by overcoming diffractionlimited technologies. The ensuing combination of “superresolution” microscopy with unbiased proteomics continues to produce neverbeforeseen gains by quantitatively addressing the distribution, interaction, turnover, and secretion of proteins in living cells. Here, we illustrate the power of this combined work flow by the example of transmembrane receptor localization at the neuronal synapse. We also discuss how dynamic analysis allows for inferences be made for cellular physiology and pathobiology.

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