og:image: citation_mjid: pnas;1801672115v1 article:published_time: 2018-05-23 og:site_name: PNAS citation_reference: citation_journal_title=Optics letters;citation_journal_abbrev=Opt Lett;citation_author=SW. Hell;citation_author=J. Wichmann;citation_title=Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy.;citation_pages=780-782;citation_volume=19;citation_year=1994;citation_issue=11;citation_pmid=19844443;citation_doi=10.1364/OL.19.000780 citation_journal_title: Proceedings of the National Academy of Sciences type: article og:description: Popular localization of single molecules through calculating the centroid of the diffraction pattern produced by molecular fluorescence on a camera is typically limited to spatiotemporal resolutions of >10 nm per >10 milliseconds. By requiring at least 10?100 times fewer detected photons and being free of bias due to molecular orientation, the localization concept called MINFLUX propels molecular tracking to the hitherto-unachievable regime of single-digit nanometer precision within substantially less than a millisecond. Our experiments herald the feasibility to detect molecular interactions and conformational changes at microsecond timescales. Compared with localization schemes solely based on evaluating patterns of molecular emission, the recently introduced single-molecule localization concept called MINFLUX and the fluorescence nanoscopies derived from it require up to orders of magnitude fewer emissions to attain single-digit nanometer resolution. Here, we demonstrate that the lower number of required fluorescence photons enables MINFLUX to detect molecular movements of a few nanometers at a temporal sampling of well below 1 millisecond. Using fluorophores attached to thermally fluctuating DNA strands as model systems, we demonstrate that measurement times as short as 400 microseconds suffice to localize fluorescent molecules with ?2-nm precision. Such performance is out of reach for popular camera-based localization by centroid calculation of emission diffraction patterns. Since theoretical limits have not been reached, our results show that emerging MINFLUX nanoscopy bears great potential for dissecting the motions of individual (macro)molecules at hitherto-unattained combinations of spatial and temporal resolution. citation_author_email: shell@mpibpc.mpg.de citation_issn: 0027-8424 citation_full_html_url: http://www.pnas.org/content/early/2018/05/22/1801672115.full citation_public_url: http://www.pnas.org/content/early/2018/05/22/1801672115 dc:title: MINFLUX monitors rapid molecular jumps with superior spatiotemporal resolution | PNAS Content-Encoding: UTF-8 citation_pdf_url: http://www.pnas.org/content/early/2018/05/22/1801672115.full.pdf citation_section: Physical Sciences citation_num_pages: 6 citation_fulltext_world_readable: citation_journal_abbrev: PNAS DC.Identifier: 10.1073/pnas.1801672115 DC.Rights: Copyright © 2018 the Author(s). Published by PNAS.. This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND). citation_author: Yvan Eilers citation_abstract_html_url: http://www.pnas.org/content/early/2018/05/22/1801672115.abstract HW.identifier: /pnas/early/2018/05/22/1801672115.atom citation_doi: 10.1073/pnas.1801672115 Content-Language: en format-detection: telephone=no Generator: Drupal 7 (http://drupal.org) citation_author_orcid: http://orcid.org/0000-0001-6209-4864 DC.AccessRights: open-access citation_publication_date: 2018/05/23 citation_title: MINFLUX monitors rapid molecular jumps with superior spatiotemporal resolution citation_author_institution: Max Planck Institute for Biophysical Chemistry citation_publisher: National Academy of Sciences citation_id: 1801672115v1 title: MINFLUX monitors rapid molecular jumps with superior spatiotemporal resolution | PNAS DC.Description: Popular localization of single molecules through calculating the centroid of the diffraction pattern produced by molecular fluorescence on a camera is typically limited to spatiotemporal resolutions of >10 nm per >10 milliseconds. By requiring at least 10?100 times fewer detected photons and being free of bias due to molecular orientation, the localization concept called MINFLUX propels molecular tracking to the hitherto-unachievable regime of single-digit nanometer precision within substantially less than a millisecond. Our experiments herald the feasibility to detect molecular interactions and conformational changes at microsecond timescales. Compared with localization schemes solely based on evaluating patterns of molecular emission, the recently introduced single-molecule localization concept called MINFLUX and the fluorescence nanoscopies derived from it require up to orders of magnitude fewer emissions to attain single-digit nanometer resolution. Here, we demonstrate that the lower number of required fluorescence photons enables MINFLUX to detect molecular movements of a few nanometers at a temporal sampling of well below 1 millisecond. Using fluorophores attached to thermally fluctuating DNA strands as model systems, we demonstrate that measurement times as short as 400 microseconds suffice to localize fluorescent molecules with ?2-nm precision. Such performance is out of reach for popular camera-based localization by centroid calculation of emission diffraction patterns. Since theoretical limits have not been reached, our results show that emerging MINFLUX nanoscopy bears great potential for dissecting the motions of individual (macro)molecules at hitherto-unattained combinations of spatial and temporal resolution. Content-Type-Hint: text/html; charset=utf-8 DC.Format: text/html DC.Publisher: National Academy of Sciences DC.Contributor: Yvan Eilers Content-Type: application/xhtml+xml; charset=UTF-8 X-Parsed-By: org.apache.tika.parser.DefaultParser og:type: article article:section: Physical Sciences citation_pmid: 29844182 citation_article_type: Research Article og:title: MINFLUX monitors rapid molecular jumps with superior spatiotemporal resolution citation_abstract:

Compared with localization schemes solely based on evaluating patterns of molecular emission, the recently introduced single-molecule localization concept called MINFLUX and the fluorescence nanoscopies derived from it require up to orders of magnitude fewer emissions to attain single-digit nanometer resolution. Here, we demonstrate that the lower number of required fluorescence photons enables MINFLUX to detect molecular movements of a few nanometers at a temporal sampling of well below 1 millisecond. Using fluorophores attached to thermally fluctuating DNA strands as model systems, we demonstrate that measurement times as short as 400 microseconds suffice to localize fluorescent molecules with ?2-nm precision. Such performance is out of reach for popular camera-based localization by centroid calculation of emission diffraction patterns. Since theoretical limits have not been reached, our results show that emerging MINFLUX nanoscopy bears great potential for dissecting the motions of individual (macro)molecules at hitherto-unattained combinations of spatial and temporal resolution.

DC.Title: MINFLUX monitors rapid molecular jumps with superior spatiotemporal resolution citation_firstpage: 201801672 viewport: initial-scale=1, maximum-scale=1, width=device-width, user-scalable=yes HW.pisa: pnas;1801672115v1 DC.Language: en DC.Date: 2018-05-29 citation_access: all category: research-article og:url: http://www.pnas.org/content/early/2018/05/22/1801672115