mEos4b photoconversion efficiency depends on laser illumination conditions used in PALM
| العنوان: | mEos4b photoconversion efficiency depends on laser illumination conditions used in PALM |
|---|---|
| المؤلفون: | Jip Wulffele, Daniel Thédié, Oleksandr Glushonkov, Dominique Bourgeois |
| المساهمون: | Institut de biologie structurale (IBS - UMR 5075), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), M4D, ANR-17-CE11-0047,Cryo-PALM,Microscopie super-résolution par localisation de molécules uniques à température cryogénique.(2017), ANR-20-CE11-0013,STABLE-FP,Nouvelle génération de protéines fluorescentes ultrastables pour la microscopie quantitative super-résolution(2020), ANR-16-CE11-0016,DIVinHD,Imagerie super-résolue de la division bactérienne(2016) |
| المصدر: | Journal of Physical Chemistry Letters Journal of Physical Chemistry Letters, 2022, 13 (22), pp.5075-5080. ⟨10.1021/acs.jpclett.2c00933⟩ |
| بيانات النشر: | Cold Spring Harbor Laboratory, 2022. |
| سنة النشر: | 2022 |
| مصطلحات موضوعية: | Microscopy, MESH: Microscopy, Light, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Lasers, Green Fluorescent Proteins, MESH: Lighting, MESH: Light, Luminescent Proteins, MESH: Green Fluorescent Proteins, MESH: Lasers, General Materials Science, MESH: Luminescent Proteins, Physical and Theoretical Chemistry, Lighting |
| الوصف: | Green-to-red photoconvertible fluorescent proteins (PCFPs) are widely employed as markers in photoactivated localization microscopy (PALM). However, their highly complex photophysical behavior complicates their usage. The fact that only a limited fraction of a PCFP ensemble can form the photoconverted state upon near-UV light illumination, termed photoconversion efficiency (PCE), lowers the achievable spatial resolution in PALM and creates undercounting errors in quantitative counting applications. Here, we show that the PCE of mEos4b is not a fixed property of this PCFP, but strongly depends on illumination conditions. Attempts to reduce long-lived blinking in red mEos4b by application of 488 nm light leads to a reduction of the PCE. Furthermore, the PCE of mEos4b strongly depends on the applied 405-nm power density. A refined photophysical model of mEos4b accounts for the observed effects, involving nonlinear green-state photobleaching upon violet light illumination favored by photon absorption by a putative radical dark state.TOC GRAPHICS |
| تدمد: | 1948-7185 |
| DOI: | 10.1101/2022.03.31.486573 |
| DOI: | 10.1021/acs.jpclett.2c00933⟩ |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::db215f72e17af5ef63f68221871d52e9 https://doi.org/10.1101/2022.03.31.486573 |
| Rights: | CLOSED |
| رقم الانضمام: | edsair.doi.dedup.....db215f72e17af5ef63f68221871d52e9 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19487185 |
|---|---|
| DOI: | 10.1101/2022.03.31.486573 |