The 2015 super-resolution microscopy roadmap

Super-resolution microscopy in Neurosciences: zoom in on synapses – Laurent

Stefan W Hell (1) , Steffen J Sahl (1) , Mark Bates (1) , Xiaowei Zhuang (2) , Rainer Heintzmann (3) , Martin J Booth (4) , Joerg Bewersdorf (5) , Gleb Shtengel (6) , Harald Hess (6) , Philipp Tinnefeld (7, 8, 9) , Alf Honigmann (10) , Stefan Jakobs (11) , Ilaria Testa (11) , Laurent Cognet (12) , Brahim Lounis (12) , Helge Ewers (13) , Simon J Davis (14) , Christian Eggeling (15) , David Klenerman (16) , Katrin I Willig (17) , Giuseppe Vicidomini (18) , Marco Castello (18) , Alberto Diaspro (18) , Thorben Cordes (19) , Steffen J Sahl , Philip Tinnefeld , David Klenerman , Katrin I Willig

1 MPI-BPC - Max Planck Institute for Biophysical Chemistry
2 HHMI - Howard Hughes Medical Institute
3 Friedrich-Schiller-Universität = Friedrich Schiller University Jena [Jena, Germany]
4 Department of Engineering Science
5 Institute for Molecular Biophysics
6 Janelia Farm Research Campus
7 Institute of Physical and Theoretical Chemistry [Braunschweig]
8 BRICS - Braunschweig Integrated Centre of Systems Biology [Braunschweig]
9 LENA - Laboratory for Emerging Nanotechnology
10 MPI-CBG - Max Planck Institute of Molecular Cell Biology and Genetics
11 Department of NanoBiophotonics [Göttingen]
12 LP2N - Laboratoire Photonique, Numérique et Nanosciences
13 Freie Universität Berlin
14 Nuffield Department of Clinical Medicine [Oxford]
15 MRC Human Immunology Unit
16 Department of Chemistry [Cambridge, UK]
17 Center for Nanoscale Microscopy and Molecular Physiology of the Brain, Göttingen, Germany
18 Nanoscopy, Nanophysics, IstitutoItaliano di Tecnologia, Via Morego, 30, 16163, Genoa, Italy
19 University of Groningen [Groningen]

Far-field optical microscopy using focused light is an important tool in a number of scientific disciplines including chemical, (bio)physical and biomedical research, particularly with respect to the study of living cells and organisms. Unfortunately, the applicability of the optical microscope is limited, since the diffraction of light imposes limitations on the spatial resolution of the image. Consequently the details of, for example, cellular protein distributions, can be visualized only to a certain extent. Fortunately, recent years have witnessed the development of 'super-resolution' far-field optical microscopy (nanoscopy) techniques such as stimulated emission depletion (STED), ground state depletion (GSD), reversible saturated optical (fluorescence) transitions (RESOLFT), photoactivation localization microscopy (PALM), stochastic optical reconstruction microscopy (STORM), structured illumination microscopy (SIM) or saturated structured illumination microscopy (SSIM), all in one way or another addressing the problem of the limited spatial resolution of far-field optical microscopy. While SIM achieves a two-fold improvement in spatial resolution compared to conventional optical microscopy, STED, RESOLFT, PALM/STORM, or SSIM have all gone beyond, pushing the limits of optical image resolution to the nanometer scale. Consequently, all super-resolution techniques open new avenues of biomedical research. Because the field is so young, the potential capabilities of different super-resolution microscopy approaches have yet to be fully explored, and uncertainties remain when considering the best choice of methodology. Thus, even for experts, the road to the future is sometimes shrouded in mist. The super-resolution optical microscopy roadmap of Journal of Physics D: Applied Physics addresses this need for clarity. It provides guidance to the outstanding questions through a collection of short review articles from experts in the field, giving a thorough discussion on the concepts underlying super-resolution optical microscopy, the potential of different approaches, the importance of label optimization (such as reversible photoswitchable proteins) and applications in which these methods will have a significant impact.

Domaines: Biophysique [physics.bio-ph]

Stefan W Hell, Steffen J Sahl, Mark Bates, Xiaowei Zhuang, Rainer Heintzmann, et al.. The 2015 super-resolution microscopy roadmap. Journal of Physics D: Applied Physics, 2015, 48 (44), pp.443001. ⟨10.1088/0022-3727/48/44/443001⟩. ⟨hal-01390051⟩