title: In vivo super-resolution RESOLFT microscopy of Drosophila melanogaster.
abstract: Despite remarkable developments in diffraction unlimited super-resolution microscopy, in vivo nanoscopy of tissues and model organisms is still not satisfactorily established and rarely realized. RESOLFT nanoscopy is particularly suited for live cell imaging because it requires relatively low light levels to overcome the diffraction barrier. Previously, we introduced the reversibly switchable fluorescent protein rsEGFP2, which facilitated fast RESOLFT nanoscopy (Grotjohann et al., 2012). In that study, as in most other nanoscopy studies, only cultivated single cells were analyzed. Here, we report on the use of rsEGFP2 for live-cell RESOLFT nanoscopy of sub-cellular structures of intact Drosophila melanogaster larvae and of resected tissues. We generated flies expressing fusion proteins of alpha-tubulin and rsEGFP2 highlighting the microtubule cytoskeleton in all cells. By focusing through the intact larval cuticle, we achieved lateral resolution of.
authors.affiliation: Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Schnorrenberg
authors.affiliation: Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
authors.forename: Tim
authors.initials: T
authors.lastname: Grotjohann
authors.affiliation: Department of Molecular Developmental Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.|Abteilung Entwicklungsbiologie, Georg-August-Universität Göttingen, Göttingen, Germany.
authors.forename: Gerd
authors.initials: G
authors.lastname: Vorbrüggen
authors.affiliation: Department of Molecular Developmental Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
authors.forename: Alf
authors.initials: A
authors.lastname: Herzig
authors.affiliation: Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
authors.forename: Stefan W
authors.initials: SW
authors.lastname: Hell
authors.affiliation: Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.|Department of Neurology, University Medical Center of Göttingen, Göttingen, Germany.
authors.forename: Stefan
authors.identifier: 0000-0002-8028-3121
authors.initials: S
authors.lastname: Jakobs
mesh_terms: D000818:Animals; D004331:Drosophila melanogaster / Q000166:cytology* / Q000648:ultrastructure*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D000069416:Intravital Microscopy / Q000379:methods*; D007814:Larva / Q000166:cytology / Q000648:ultrastructure; D013194:Staining and Labeling
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: D. melanogaster; RESOLFT; cell biology; imaging; in vivo; mircrotubule cytoskeleton; nanoscopy; super resolution