MINSTED nanoscopy enters the Ångström localization range
Super-resolution techniques have achieved localization precisions in the nanometer regime. Here we report all-optical, room temperature localization of fluorophores with precision in the Ångström range. We built on the concept of MINSTED nanoscopy where precision is increased by encircling the fluor...
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| Veröffentlicht in: | Nature biotechnology 2023-04, Vol.41 (4), p.569-576 |
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| creator | Weber, Michael von der Emde, Henrik Leutenegger, Marcel Gunkel, Philip Sambandan, Sivakumar Khan, Taukeer A. Keller-Findeisen, Jan Cordes, Volker C. Hell, Stefan W. |
| description | Super-resolution techniques have achieved localization precisions in the nanometer regime. Here we report all-optical, room temperature localization of fluorophores with precision in the Ångström range. We built on the concept of MINSTED nanoscopy where precision is increased by encircling the fluorophore with the low-intensity central region of a stimulated emission depletion (STED) donut beam while constantly increasing the absolute donut power. By blue-shifting the STED beam and separating fluorophores by on/off switching, individual fluorophores bound to a DNA strand are localized with
σ
= 4.7 Å, corresponding to a fraction of the fluorophore size, with only 2,000 detected photons. MINSTED fluorescence nanoscopy with single-digit nanometer resolution is exemplified by imaging nuclear pore complexes and the distribution of nuclear lamin in mammalian cells labeled by transient DNA hybridization. Because our experiments yield a localization precision
σ
= 2.3 Å, estimated for 10,000 detected photons, we anticipate that MINSTED will open up new areas of application in the study of macromolecular complexes in cells.
Sub-nanometer localization precision is achieved with all-optical microscopy. |
| doi_str_mv | 10.1038/s41587-022-01519-4 |
| format | Article |
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σ
= 4.7 Å, corresponding to a fraction of the fluorophore size, with only 2,000 detected photons. MINSTED fluorescence nanoscopy with single-digit nanometer resolution is exemplified by imaging nuclear pore complexes and the distribution of nuclear lamin in mammalian cells labeled by transient DNA hybridization. Because our experiments yield a localization precision
σ
= 2.3 Å, estimated for 10,000 detected photons, we anticipate that MINSTED will open up new areas of application in the study of macromolecular complexes in cells.
Sub-nanometer localization precision is achieved with all-optical microscopy.</description><identifier>ISSN: 1087-0156</identifier><identifier>EISSN: 1546-1696</identifier><identifier>DOI: 10.1038/s41587-022-01519-4</identifier><identifier>PMID: 36344840</identifier><language>eng</language><publisher>New York: Nature Publishing Group US</publisher><subject>631/1647/328/2238 ; 639/624/1107/328/2238 ; Agriculture ; Animals ; Bioinformatics ; Biomedical and Life Sciences ; Biomedical Engineering/Biotechnology ; Biomedicine ; Biotechnology ; Chemical compounds ; Deoxyribonucleic acid ; Depletion ; DNA ; Fluorescence ; Fluorescent Dyes ; Fluorophores ; Hybridization ; Life Sciences ; Light microscopy ; Localization ; Macromolecules ; Mammalian cells ; Mammals ; Microscopy, Fluorescence - methods ; Nuclear pores ; Optical microscopy ; Photons ; Room temperature ; Stimulated emission</subject><ispartof>Nature biotechnology, 2023-04, Vol.41 (4), p.569-576</ispartof><rights>The Author(s) 2022</rights><rights>2022. The Author(s).</rights><rights>The Author(s) 2022. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c475t-671fd8dc8fe7a76e5637d8684497762f07f8bd3ec5d5379cc84fa7fc5a03ef6a3</citedby><cites>FETCH-LOGICAL-c475t-671fd8dc8fe7a76e5637d8684497762f07f8bd3ec5d5379cc84fa7fc5a03ef6a3</cites><orcidid>0000-0003-4287-9482 ; 0000-0002-3188-6612 ; 0000-0002-9638-5077 ; 0000-0001-9680-7543 ; 0000-0003-4731-0148</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/s41587-022-01519-4$$EPDF$$P50$$Gspringer$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/s41587-022-01519-4$$EHTML$$P50$$Gspringer$$Hfree_for_read</linktohtml><link.rule.ids>230,314,776,780,881,27903,27904,41467,42536,51297</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/36344840$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Weber, Michael</creatorcontrib><creatorcontrib>von der Emde, Henrik</creatorcontrib><creatorcontrib>Leutenegger, Marcel</creatorcontrib><creatorcontrib>Gunkel, Philip</creatorcontrib><creatorcontrib>Sambandan, Sivakumar</creatorcontrib><creatorcontrib>Khan, Taukeer A.</creatorcontrib><creatorcontrib>Keller-Findeisen, Jan</creatorcontrib><creatorcontrib>Cordes, Volker C.</creatorcontrib><creatorcontrib>Hell, Stefan W.</creatorcontrib><title>MINSTED nanoscopy enters the Ångström localization range</title><title>Nature biotechnology</title><addtitle>Nat Biotechnol</addtitle><addtitle>Nat Biotechnol</addtitle><description>Super-resolution techniques have achieved localization precisions in the nanometer regime. Here we report all-optical, room temperature localization of fluorophores with precision in the Ångström range. We built on the concept of MINSTED nanoscopy where precision is increased by encircling the fluorophore with the low-intensity central region of a stimulated emission depletion (STED) donut beam while constantly increasing the absolute donut power. By blue-shifting the STED beam and separating fluorophores by on/off switching, individual fluorophores bound to a DNA strand are localized with
σ
= 4.7 Å, corresponding to a fraction of the fluorophore size, with only 2,000 detected photons. MINSTED fluorescence nanoscopy with single-digit nanometer resolution is exemplified by imaging nuclear pore complexes and the distribution of nuclear lamin in mammalian cells labeled by transient DNA hybridization. Because our experiments yield a localization precision
σ
= 2.3 Å, estimated for 10,000 detected photons, we anticipate that MINSTED will open up new areas of application in the study of macromolecular complexes in cells.
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enters the Ångström localization range</atitle><jtitle>Nature biotechnology</jtitle><stitle>Nat Biotechnol</stitle><addtitle>Nat Biotechnol</addtitle><date>2023-04-01</date><risdate>2023</risdate><volume>41</volume><issue>4</issue><spage>569</spage><epage>576</epage><pages>569-576</pages><issn>1087-0156</issn><eissn>1546-1696</eissn><abstract>Super-resolution techniques have achieved localization precisions in the nanometer regime. Here we report all-optical, room temperature localization of fluorophores with precision in the Ångström range. We built on the concept of MINSTED nanoscopy where precision is increased by encircling the fluorophore with the low-intensity central region of a stimulated emission depletion (STED) donut beam while constantly increasing the absolute donut power. By blue-shifting the STED beam and separating fluorophores by on/off switching, individual fluorophores bound to a DNA strand are localized with
σ
= 4.7 Å, corresponding to a fraction of the fluorophore size, with only 2,000 detected photons. MINSTED fluorescence nanoscopy with single-digit nanometer resolution is exemplified by imaging nuclear pore complexes and the distribution of nuclear lamin in mammalian cells labeled by transient DNA hybridization. Because our experiments yield a localization precision
σ
= 2.3 Å, estimated for 10,000 detected photons, we anticipate that MINSTED will open up new areas of application in the study of macromolecular complexes in cells.
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| subjects | 631/1647/328/2238 639/624/1107/328/2238 Agriculture Animals Bioinformatics Biomedical and Life Sciences Biomedical Engineering/Biotechnology Biomedicine Biotechnology Chemical compounds Deoxyribonucleic acid Depletion DNA Fluorescence Fluorescent Dyes Fluorophores Hybridization Life Sciences Light microscopy Localization Macromolecules Mammalian cells Mammals Microscopy, Fluorescence - methods Nuclear pores Optical microscopy Photons Room temperature Stimulated emission |
| title | MINSTED nanoscopy enters the Ångström localization range |
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