Dual-color fluorescence lifetime correlation spectroscopy to quantify protein-protein interactions in live cell.
Dual-color fluorescence correlation spectroscopy is an interesting method to quantify protein interaction in living cells. But, when performing these experiments, one must compensate for a known spectral bleed through artifact that corrupts cross-correlation data. In this article, problems with cros...
| Main Authors: | , , , |
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| Format: | Journal article |
| Language: | English |
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2011
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| _version_ | 1826293494176546816 |
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| author | Padilla-Parra, S Audugé, N Coppey-Moisan, M Tramier, M |
| author_facet | Padilla-Parra, S Audugé, N Coppey-Moisan, M Tramier, M |
| author_sort | Padilla-Parra, S |
| collection | OXFORD |
| description | Dual-color fluorescence correlation spectroscopy is an interesting method to quantify protein interaction in living cells. But, when performing these experiments, one must compensate for a known spectral bleed through artifact that corrupts cross-correlation data. In this article, problems with crosstalk were overcome with an approach based on fluorescence lifetime correlation spectroscopy (FLCS). We show that FLCS applied to dual-color EGFP and mCherry cross-correlation allows the determination of protein-protein interactions in living cells without the need of spectral bleed through calibration. The methodology was validated by using EGFP-mCherry tandem in comparison with coexpressed EGFP and mCherry in live cell. The dual-color FLCS experimental procedure where the different laser intensities do not have to be controlled during experiment is really very helpful to study quantitatively protein interactions in live sample. |
| first_indexed | 2024-03-07T03:30:58Z |
| format | Journal article |
| id | oxford-uuid:bab37a58-a485-467f-8ced-84ff7f353122 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T03:30:58Z |
| publishDate | 2011 |
| record_format | dspace |
| spelling | oxford-uuid:bab37a58-a485-467f-8ced-84ff7f3531222022-03-27T05:11:39ZDual-color fluorescence lifetime correlation spectroscopy to quantify protein-protein interactions in live cell.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:bab37a58-a485-467f-8ced-84ff7f353122EnglishSymplectic Elements at Oxford2011Padilla-Parra, SAudugé, NCoppey-Moisan, MTramier, MDual-color fluorescence correlation spectroscopy is an interesting method to quantify protein interaction in living cells. But, when performing these experiments, one must compensate for a known spectral bleed through artifact that corrupts cross-correlation data. In this article, problems with crosstalk were overcome with an approach based on fluorescence lifetime correlation spectroscopy (FLCS). We show that FLCS applied to dual-color EGFP and mCherry cross-correlation allows the determination of protein-protein interactions in living cells without the need of spectral bleed through calibration. The methodology was validated by using EGFP-mCherry tandem in comparison with coexpressed EGFP and mCherry in live cell. The dual-color FLCS experimental procedure where the different laser intensities do not have to be controlled during experiment is really very helpful to study quantitatively protein interactions in live sample. |
| spellingShingle | Padilla-Parra, S Audugé, N Coppey-Moisan, M Tramier, M Dual-color fluorescence lifetime correlation spectroscopy to quantify protein-protein interactions in live cell. |
| title | Dual-color fluorescence lifetime correlation spectroscopy to quantify protein-protein interactions in live cell. |
| title_full | Dual-color fluorescence lifetime correlation spectroscopy to quantify protein-protein interactions in live cell. |
| title_fullStr | Dual-color fluorescence lifetime correlation spectroscopy to quantify protein-protein interactions in live cell. |
| title_full_unstemmed | Dual-color fluorescence lifetime correlation spectroscopy to quantify protein-protein interactions in live cell. |
| title_short | Dual-color fluorescence lifetime correlation spectroscopy to quantify protein-protein interactions in live cell. |
| title_sort | dual color fluorescence lifetime correlation spectroscopy to quantify protein protein interactions in live cell |
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