Robin Van den Eynde, Fabian Hertel, Sergey Abakumov, Bartosz Krajnik, Siewert Hugelier, Alexander Auer, Joschka Hellmeier, Thomas Schlichthaerle, Rachel M. Grattan, Diane S. Lidke, Ralf Jungmann, Marcel Leutenegger, Wim Vandenberg, Peter Dedecker
{"title":"Simultaneous multicolor fluorescence imaging using PSF splitting","authors":"Robin Van den Eynde, Fabian Hertel, Sergey Abakumov, Bartosz Krajnik, Siewert Hugelier, Alexander Auer, Joschka Hellmeier, Thomas Schlichthaerle, Rachel M. Grattan, Diane S. Lidke, Ralf Jungmann, Marcel Leutenegger, Wim Vandenberg, Peter Dedecker","doi":"10.1038/s41592-024-02383-7","DOIUrl":null,"url":null,"abstract":"We present a way to encode more information in fluorescence imaging by splitting the original point spread function (PSF), which offers broadband operation and compatibility with other PSF engineering modalities and existing analysis tools. We demonstrate the approach using the ‘Circulator’, an add-on that encodes the fluorophore emission band into the PSF, enabling simultaneous multicolor super-resolution and single-molecule microscopy using essentially the full field of view. Point spread function (PSF) splitting with the ‘Circulator’, which encodes the fluorophore emission band into the PSF, improves the information content of fluorescence microscopy and enables improved super-resolution imaging and single-particle tracking.","PeriodicalId":18981,"journal":{"name":"Nature Methods","volume":null,"pages":null},"PeriodicalIF":36.1000,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Nature Methods","FirstCategoryId":"99","ListUrlMain":"https://www.nature.com/articles/s41592-024-02383-7","RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
Abstract
We present a way to encode more information in fluorescence imaging by splitting the original point spread function (PSF), which offers broadband operation and compatibility with other PSF engineering modalities and existing analysis tools. We demonstrate the approach using the ‘Circulator’, an add-on that encodes the fluorophore emission band into the PSF, enabling simultaneous multicolor super-resolution and single-molecule microscopy using essentially the full field of view. Point spread function (PSF) splitting with the ‘Circulator’, which encodes the fluorophore emission band into the PSF, improves the information content of fluorescence microscopy and enables improved super-resolution imaging and single-particle tracking.
期刊介绍：
Nature Methods is a monthly journal that focuses on publishing innovative methods and substantial enhancements to fundamental life sciences research techniques. Geared towards a diverse, interdisciplinary readership of researchers in academia and industry engaged in laboratory work, the journal offers new tools for research and emphasizes the immediate practical significance of the featured work. It publishes primary research papers and reviews recent technical and methodological advancements, with a particular interest in primary methods papers relevant to the biological and biomedical sciences. This includes methods rooted in chemistry with practical applications for studying biological problems.
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