% IMPORTANT: The following is UTF-8 encoded.  This means that in the presence
% of non-ASCII characters, it will not work with BibTeX 0.99 or older.
% Instead, you should use an up-to-date BibTeX implementation like “bibtex8” or
% “biber”.

@ARTICLE{Masch:136959,
      author       = {J.-M. Masch and H. Steffens and J. Fischer$^*$ and J.
                      Engelhardt$^*$ and J. Hubrich and J. Keller-Findeisen and E.
                      D'Este and N. T. Urban and S. G. N. Grant and S. J. Sahl and
                      D. Kamin and S. Hell$^*$},
      title        = {{R}obust nanoscopy of a synaptic protein in living mice by
                      organic-fluorophore labeling.},
      journal      = {Proceedings of the National Academy of Sciences of the
                      United States of America},
      volume       = {115},
      number       = {34},
      issn         = {1091-6490},
      address      = {Washington, DC},
      publisher    = {National Acad. of Sciences},
      reportid     = {DKFZ-2018-01388},
      pages        = {E8047 - E8056},
      year         = {2018},
      abstract     = {Extending superresolution fluorescence microscopy to living
                      animals has remained a challenging frontier ever since the
                      first demonstration of STED (stimulated emission depletion)
                      nanoscopy in the mouse visual cortex. The use of fluorescent
                      proteins (FPs) in in vivo STED analyses has been limiting
                      available fluorescence photon budgets and attainable image
                      contrasts, in particular for far-red FPs. This has so far
                      precluded the definition of subtle details in protein
                      arrangements at sufficient signal-to-noise ratio.
                      Furthermore, imaging with longer wavelengths holds promise
                      for reducing photostress. Here, we demonstrate that a
                      strategy based on enzymatic self-labeling of the HaloTag
                      fusion protein by high-performance synthetic fluorophore
                      labels provides a robust avenue to superior in vivo analysis
                      with STED nanoscopy in the far-red spectral range. We
                      illustrate our approach by mapping the nanoscale
                      distributions of the abundant scaffolding protein PSD95 at
                      the postsynaptic membrane of excitatory synapses in living
                      mice. With silicon-rhodamine as the reporter fluorophore, we
                      present imaging with high contrast and low background down
                      to ∼70-nm lateral resolution in the visual cortex at
                      ≤25-µm depth. This approach allowed us to identify and
                      characterize the diversity of PSD95 scaffolds in vivo.
                      Besides small round/ovoid shapes, a substantial fraction of
                      scaffolds exhibited a much more complex spatial
                      organization. This highly inhomogeneous, spatially extended
                      PSD95 distribution within the disk-like postsynaptic
                      density, featuring intricate perforations, has not been
                      highlighted in cell- or tissue-culture experiments.
                      Importantly, covisualization of the corresponding spine
                      morphologies enabled us to contextualize the diverse PSD95
                      patterns within synapses of different orientations and
                      sizes.},
      cin          = {E190},
      ddc          = {000},
      cid          = {I:(DE-He78)E190-20160331},
      pnm          = {315 - Imaging and radiooncology (POF3-315)},
      pid          = {G:(DE-HGF)POF3-315},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:30082388},
      pmc          = {pmc:PMC6112726},
      doi          = {10.1073/pnas.1807104115},
      url          = {https://inrepo02.dkfz.de/record/136959},
}