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  • 1
    Publication Date: 2011-09-13
    Description: Lens-based optical microscopy failed to discern fluorescent features closer than 200 nm for decades, but the recent breaking of the diffraction resolution barrier by sequentially switching the fluorescence capability of adjacent features on and off is making nanoscale imaging routine. Reported fluorescence nanoscopy variants switch these features either with intense beams at defined positions or randomly, molecule by molecule. Here we demonstrate an optical nanoscopy that records raw data images from living cells and tissues with low levels of light. This advance has been facilitated by the generation of reversibly switchable enhanced green fluorescent protein (rsEGFP), a fluorescent protein that can be reversibly photoswitched more than a thousand times. Distributions of functional rsEGFP-fusion proteins in living bacteria and mammalian cells are imaged at 〈40-nanometre resolution. Dendritic spines in living brain slices are super-resolved with about a million times lower light intensities than before. The reversible switching also enables all-optical writing of features with subdiffraction size and spacings, which can be used for data storage.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Grotjohann, Tim -- Testa, Ilaria -- Leutenegger, Marcel -- Bock, Hannes -- Urban, Nicolai T -- Lavoie-Cardinal, Flavie -- Willig, Katrin I -- Eggeling, Christian -- Jakobs, Stefan -- Hell, Stefan W -- England -- Nature. 2011 Sep 11;478(7368):204-8. doi: 10.1038/nature10497.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Gottingen, Germany.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/21909116" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; Brain/cytology ; Cell Line ; Cell Survival ; Dendrites ; Equipment Reuse ; Escherichia coli/metabolism ; Green Fluorescent Proteins/chemistry/genetics/*metabolism ; Light ; Microscopy, Fluorescence/*methods ; Nanotechnology/methods ; Optics and Photonics/*methods ; Photobleaching
    Print ISSN: 0028-0836
    Electronic ISSN: 1476-4687
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
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  • 2
    Publication Date: 2016-04-30
    Description: Li et al (Research Articles, 28 August 2015, aab3500) purport to present solutions to long-standing challenges in live-cell microscopy, reporting relatively fast acquisition times in conjunction with improved image resolution. We question the methods' reliability to visualize specimen features at sub-100-nanometer scales, because the mandatory mathematical processing of the recorded data leads to artifacts that are either difficult or impossible to disentangle from real features. We are also concerned about the chosen approach of subjectively comparing images from different super-resolution methods, as opposed to using quantitative measures.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Sahl, Steffen J -- Balzarotti, Francisco -- Keller-Findeisen, Jan -- Leutenegger, Marcel -- Westphal, Volker -- Egner, Alexander -- Lavoie-Cardinal, Flavie -- Chmyrov, Andriy -- Grotjohann, Tim -- Jakobs, Stefan -- New York, N.Y. -- Science. 2016 Apr 29;352(6285):527. doi: 10.1126/science.aad7983. Epub 2016 Apr 28.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Max Planck Institute for Biophysical Chemistry, Department of NanoBiophotonics, Am Fassberg 11, 37077 Gottingen, Germany. ssahl@mpibpc.mpg.de fbalzar@mpibpc.mpg.de sjakobs@mpibpc.mpg.de. ; Max Planck Institute for Biophysical Chemistry, Department of NanoBiophotonics, Am Fassberg 11, 37077 Gottingen, Germany. ; Laser Laboratory Gottingen, Hans-Adolf-Krebs-Weg 1, 37077 Gottingen, Germany. ; Institut Universitaire en Sante Mentale de Quebec, Cellular and Molecular Neuroscience Research Axis, 2601 Chemin de la Canardiere, Quebec, G1J 2G3, Canada. ; Helmholtz Zentrum Munchen, Institute of Biological and Medical Imaging, Ingolstadter Landstrasse 1, 85764 Neuherberg, Germany. Technische Universitat Munchen, Chair for Biological Imaging, Ismaningerstrasse 22, 81675 Munchen, Germany. ; Max Planck Institute for Biophysical Chemistry, Department of NanoBiophotonics, Am Fassberg 11, 37077 Gottingen, Germany. University of Gottingen Medical Faculty, Department of Neurology, Robert-Koch-Str. 40, 37075 Gottingen, Germany. ssahl@mpibpc.mpg.de fbalzar@mpibpc.mpg.de sjakobs@mpibpc.mpg.de.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/27126030" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; Cytoskeleton/*ultrastructure ; *Endocytosis ; Imaging, Three-Dimensional/*methods ; Microscopy, Fluorescence/*methods ; Organelles/*ultrastructure
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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