Resolution doubling in 3D-STORM imaging through improved buffers
Super-resolution imaging methods have revolutionized fluorescence microscopy by revealing the nanoscale organization of labeled proteins. In particular, single-molecule methods such as Stochastic Optical Reconstruction Microscopy (STORM) provide resolutions down to a few tens of nanometers by exploi...
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| Veröffentlicht in: | PloS one 2013-07, Vol.8 (7), p.e69004-e69004 |
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| creator | Olivier, Nicolas Keller, Debora Gönczy, Pierre Manley, Suliana |
| description | Super-resolution imaging methods have revolutionized fluorescence microscopy by revealing the nanoscale organization of labeled proteins. In particular, single-molecule methods such as Stochastic Optical Reconstruction Microscopy (STORM) provide resolutions down to a few tens of nanometers by exploiting the cycling of dyes between fluorescent and non-fluorescent states to obtain a sparse population of emitters and precisely localizing them individually. This cycling of dyes is commonly induced by adding different chemicals, which are combined to create a STORM buffer. Despite their importance, the composition of these buffers has scarcely evolved since they were first introduced, fundamentally limiting what can be resolved with STORM. By identifying a new chemical suitable for STORM and optimizing the buffer composition for Alexa-647, we significantly increased the number of photons emitted per cycle by each dye, providing a simple means to enhance the resolution of STORM independently of the optical setup used. Using this buffer to perform 3D-STORM on biological samples, we obtained images with better than 10 nanometer lateral and 30 nanometer axial resolution. |
| doi_str_mv | 10.1371/journal.pone.0069004 |
| format | Article |
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Using this buffer to perform 3D-STORM on biological samples, we obtained images with better than 10 nanometer lateral and 30 nanometer axial resolution.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0069004</identifier><identifier>PMID: 23874848</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Animals ; Biological properties ; Biological samples ; Biology ; Biophysics ; Buffers ; Cercopithecus aethiops ; Chemistry ; COS Cells ; Cycles ; Dyes ; Emitters ; Experiments ; Fluorescence ; Fluorescence microscopy ; Hydrogen-Ion Concentration ; Image resolution ; Laboratories ; Life sciences ; Localization ; Medical research ; Microscopy ; Microscopy, Fluorescence - methods ; Photons ; Physics ; Proteins ; Proteins - metabolism ; Stochasticity ; Storms</subject><ispartof>PloS one, 2013-07, Vol.8 (7), p.e69004-e69004</ispartof><rights>COPYRIGHT 2013 Public Library of Science</rights><rights>2013 Olivier et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Olivier, Nicolas</au><au>Keller, Debora</au><au>Gönczy, Pierre</au><au>Manley, Suliana</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Resolution doubling in 3D-STORM imaging through improved buffers</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2013-07-17</date><risdate>2013</risdate><volume>8</volume><issue>7</issue><spage>e69004</spage><epage>e69004</epage><pages>e69004-e69004</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Super-resolution imaging methods have revolutionized fluorescence microscopy by revealing the nanoscale organization of labeled proteins. In particular, single-molecule methods such as Stochastic Optical Reconstruction Microscopy (STORM) provide resolutions down to a few tens of nanometers by exploiting the cycling of dyes between fluorescent and non-fluorescent states to obtain a sparse population of emitters and precisely localizing them individually. This cycling of dyes is commonly induced by adding different chemicals, which are combined to create a STORM buffer. Despite their importance, the composition of these buffers has scarcely evolved since they were first introduced, fundamentally limiting what can be resolved with STORM. By identifying a new chemical suitable for STORM and optimizing the buffer composition for Alexa-647, we significantly increased the number of photons emitted per cycle by each dye, providing a simple means to enhance the resolution of STORM independently of the optical setup used. Using this buffer to perform 3D-STORM on biological samples, we obtained images with better than 10 nanometer lateral and 30 nanometer axial resolution.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>23874848</pmid><doi>10.1371/journal.pone.0069004</doi><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Biological properties Biological samples Biology Biophysics Buffers Cercopithecus aethiops Chemistry COS Cells Cycles Dyes Emitters Experiments Fluorescence Fluorescence microscopy Hydrogen-Ion Concentration Image resolution Laboratories Life sciences Localization Medical research Microscopy Microscopy, Fluorescence - methods Photons Physics Proteins Proteins - metabolism Stochasticity Storms |
| title | Resolution doubling in 3D-STORM imaging through improved buffers |
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