دورية أكاديمية
SAIBR: a simple, platform-independent method for spectral autofluorescence correction.
| العنوان: | SAIBR: a simple, platform-independent method for spectral autofluorescence correction. |
|---|---|
| المؤلفون: | Rodrigues NTL; Francis Crick Institute, London NW1 1AT, UK., Bland T; Francis Crick Institute, London NW1 1AT, UK.; Institute for the Physics of Living Systems, University College London, London WC1E 6BT, UK., Borrego-Pinto J; Francis Crick Institute, London NW1 1AT, UK., Ng K; Francis Crick Institute, London NW1 1AT, UK.; Institute for the Physics of Living Systems, University College London, London WC1E 6BT, UK., Hirani N; Francis Crick Institute, London NW1 1AT, UK., Gu Y; Francis Crick Institute, London NW1 1AT, UK.; Randall Centre for Cell and Molecular Biophysics, School of Basic and Medical Biosciences, King's College London, London SE1 1UL, UK., Foo S; Francis Crick Institute, London NW1 1AT, UK.; Randall Centre for Cell and Molecular Biophysics, School of Basic and Medical Biosciences, King's College London, London SE1 1UL, UK., Goehring NW; Francis Crick Institute, London NW1 1AT, UK.; Institute for the Physics of Living Systems, University College London, London WC1E 6BT, UK. |
| المصدر: | Development (Cambridge, England) [Development] 2022 Jul 15; Vol. 149 (14). Date of Electronic Publication: 2022 Jul 14. |
| نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Company Of Biologists Limited Country of Publication: England NLM ID: 8701744 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1477-9129 (Electronic) Linking ISSN: 09501991 NLM ISO Abbreviation: Development Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Cambridge Eng : Company Of Biologists Limited Original Publication: [Cambridge] : Company of Biologists, [c1987- |
| مواضيع طبية MeSH: | Caenorhabditis elegans* , Proteins*, Animals ; Fluorescence ; Fluorescent Dyes ; Genes, Reporter |
| مستخلص: | Biological systems are increasingly viewed through a quantitative lens that demands accurate measures of gene expression and local protein concentrations. CRISPR/Cas9 gene tagging has enabled increased use of fluorescence to monitor proteins at or near endogenous levels under native regulatory control. However, owing to typically lower expression levels, experiments using endogenously tagged genes run into limits imposed by autofluorescence (AF). AF is often a particular challenge in wavelengths occupied by commonly used fluorescent proteins (GFP, mNeonGreen). Stimulated by our work in C. elegans, we describe and validate Spectral Autofluorescence Image Correction By Regression (SAIBR), a simple platform-independent protocol and FIJI plug-in to correct for autofluorescence using standard filter sets and illumination conditions. Validated for use in C. elegans embryos, starfish oocytes and fission yeast, SAIBR is ideal for samples with a single dominant AF source; it achieves accurate quantitation of fluorophore signal, and enables reliable detection and quantification of even weakly expressed proteins. Thus, SAIBR provides a highly accessible low-barrier way to incorporate AF correction as standard for researchers working on a broad variety of cell and developmental systems. Competing Interests: Competing interests The authors declare no competing or financial interests. (© 2022. Published by The Company of Biologists Ltd.) |
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| معلومات مُعتمدة: | FC001086 United Kingdom WT_ Wellcome Trust; P40 OD010440 United States OD NIH HHS; 220790/Z/20/Z United Kingdom WT_ Wellcome Trust; BB/T000481/1 United Kingdom BB_ Biotechnology and Biological Sciences Research Council; FC001086 United Kingdom MRC_ Medical Research Council; FC001086 United Kingdom CRUK_ Cancer Research UK |
| فهرسة مساهمة: | Keywords: C. elegans; S. pombe; Autofluorescence correction; Fiji plug-in; Starfish |
| المشرفين على المادة: | 0 (Fluorescent Dyes) 0 (Proteins) |
| تواريخ الأحداث: | Date Created: 20220617 Date Completed: 20220718 Latest Revision: 20230816 |
| رمز التحديث: | 20231215 |
| مُعرف محوري في PubMed: | PMC9445497 |
| DOI: | 10.1242/dev.200545 |
| PMID: | 35713287 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1477-9129 |
|---|---|
| DOI: | 10.1242/dev.200545 |