Super-resolution imaging and estimation of protein copy numbers at single synapses with DNA-point accumulation for imaging in nanoscale topography

利用 DNA 点积累对单个突触进行超分辨率成像和蛋白质拷贝数估计,以实现纳米级拓扑结构成像

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作者:Carolin Böger, Anne-Sophie Hafner, Thomas Schlichthärle, Maximilian T Strauss, Sebastian Malkusch, Ulrike Endesfelder, Ralf Jungmann, Erin M Schuman, Mike Heilemann

Abstract

In the brain, the strength of each individual synapse is defined by the complement of proteins present or the "local proteome." Activity-dependent changes in synaptic strength are the result of changes in this local proteome and posttranslational protein modifications. Although most synaptic proteins have been identified, we still know little about protein copy numbers in individual synapses and variations between synapses. We use DNA-point accumulation for imaging in nanoscale topography as a single-molecule super-resolution imaging technique to visualize and quantify protein copy numbers in single synapses. The imaging technique provides near-molecular spatial resolution, is unaffected by photobleaching, enables imaging of large field of views, and provides quantitative molecular information. We demonstrate these benefits by accessing copy numbers of surface AMPA-type receptors at single synapses of rat hippocampal neurons along dendritic segments.

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