{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:48:46Z","timestamp":1760147326736,"version":"build-2065373602"},"reference-count":37,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2023,1,23]],"date-time":"2023-01-23T00:00:00Z","timestamp":1674432000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the Deutsche Forschungsgemeinschaft (DFG) under Germany\u2019s Excellence Strategy\u2014EXC","award":["2067\/1-390729940"],"award-info":[{"award-number":["2067\/1-390729940"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nanomaterials"],"abstract":"<jats:p>With the advent of fluorescence superresolution microscopy, nano-sized structures can be imaged with a previously unprecedented accuracy. Therefore, it is rapidly gaining importance as an analytical tool in the life sciences and beyond. However, the images obtained so far lack an absolute scale in terms of fluorophore numbers. Here, we use, for the first time, a detailed statistical model of the temporal imaging process which relies on a hidden Markov model operating on two timescales. This allows us to extract this information from the raw data without additional calibration measurements. We show this on the basis of added data from experiments on single Alexa 647 molecules as well as GSDIM\/dSTORM measurements on DNA origami structures with a known number of labeling positions.<\/jats:p>","DOI":"10.3390\/nano13030459","type":"journal-article","created":{"date-parts":[[2023,1,23]],"date-time":"2023-01-23T04:19:22Z","timestamp":1674447562000},"page":"459","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Towards Unbiased Fluorophore Counting in Superresolution Fluorescence Microscopy"],"prefix":"10.3390","volume":"13","author":[{"given":"Oskar","family":"Laitenberger","sequence":"first","affiliation":[{"name":"Department of Optical Nanoscopy, Institut f\u00fcr Nanophotonik e.V., 37077 G\u00f6ttingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Timo","family":"Aspelmeier","sequence":"additional","affiliation":[{"name":"Institute for Mathematical Stochastics, Georg-August-University of G\u00f6ttingen, 37073 G\u00f6ttingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thomas","family":"Staudt","sequence":"additional","affiliation":[{"name":"Institute for Mathematical Stochastics, Georg-August-University of G\u00f6ttingen, 37073 G\u00f6ttingen, Germany"},{"name":"Cluster of Excellence \u201cMultiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells\u201d (MBExC), University of G\u00f6ttingen, 37075 G\u00f6ttingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claudia","family":"Geisler","sequence":"additional","affiliation":[{"name":"Department of Optical Nanoscopy, Institut f\u00fcr Nanophotonik e.V., 37077 G\u00f6ttingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9181-9331","authenticated-orcid":false,"given":"Axel","family":"Munk","sequence":"additional","affiliation":[{"name":"Institute for Mathematical Stochastics, Georg-August-University of G\u00f6ttingen, 37073 G\u00f6ttingen, Germany"},{"name":"Cluster of Excellence \u201cMultiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells\u201d (MBExC), University of G\u00f6ttingen, 37075 G\u00f6ttingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5248-3858","authenticated-orcid":false,"given":"Alexander","family":"Egner","sequence":"additional","affiliation":[{"name":"Department of Optical Nanoscopy, Institut f\u00fcr Nanophotonik e.V., 37077 G\u00f6ttingen, Germany"},{"name":"Cluster of Excellence \u201cMultiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells\u201d (MBExC), University of G\u00f6ttingen, 37075 G\u00f6ttingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1126\/science.1124618","article-title":"The Fluorescent Toolbox for Assessing Protein Location and Function","volume":"312","author":"Giepmans","year":"2006","journal-title":"Science"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"697","DOI":"10.1021\/bi0118512","article-title":"Biological and Chemical Applications of Fluorescence Correlation Spectroscopy: A Review","volume":"41","author":"Hess","year":"2002","journal-title":"Biochemistry"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"124601","DOI":"10.1088\/0034-4885\/78\/12\/124601","article-title":"A review of progress in single particle tracking: From methods to biophysical insights","volume":"78","author":"Manzo","year":"2015","journal-title":"Rep. 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