Assessment of Gate Width Size on Lifetime-Based Förster Resonance Energy Transfer Parameter Estimation
Förster Resonance Energy Transfer (FRET) enables the observation of interactions at the nanoscale level through the use of fluorescence optical imaging techniques. In FRET, fluorescence lifetime imaging can be used to quantify the fluorescence lifetime changes of the donor molecule, which are associ...
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| Format: | Article |
| Language: | English |
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MDPI AG
2015-09-01
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| Series: | Photonics |
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| Online Access: | http://www.mdpi.com/2304-6732/2/4/1027 |
| _version_ | 1831632207120171008 |
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| author | Sez-Jade Chen Nattawut Sinsuebphon Xavier Intes |
| author_facet | Sez-Jade Chen Nattawut Sinsuebphon Xavier Intes |
| author_sort | Sez-Jade Chen |
| collection | DOAJ |
| description | Förster Resonance Energy Transfer (FRET) enables the observation of interactions at the nanoscale level through the use of fluorescence optical imaging techniques. In FRET, fluorescence lifetime imaging can be used to quantify the fluorescence lifetime changes of the donor molecule, which are associated with proximity between acceptor and donor molecules. Among the FRET parameters derived from fluorescence lifetime imaging, the percentage of donor that interacts with the acceptor (in proximity) can be estimated via model-based fitting. However, estimation of the lifetime parameters can be affected by the acquisition parameters such as the temporal characteristics of the imaging system. Herein, we investigate the effect of various gate widths on the accuracy of estimation of FRET parameters with focus on the near-infrared spectral window. Experiments were performed in silico, in vitro, and in vivo with gate width sizes ranging from 300 ps to 1000 ps in intervals of 100 ps. For all cases, the FRET parameters were retrieved accurately and the imaging acquisition time was decreased three-fold. These results indicate that increasing the gate width up to 1000 ps still allows for accurate quantification of FRET interactions even in the case of short lifetimes such as those encountered with near-infrared FRET pairs. |
| first_indexed | 2024-12-19T05:14:52Z |
| format | Article |
| id | doaj.art-132af7d48d7f4283b629bc40cdb449ab |
| institution | Directory Open Access Journal |
| issn | 2304-6732 |
| language | English |
| last_indexed | 2024-12-19T05:14:52Z |
| publishDate | 2015-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Photonics |
| spelling | doaj.art-132af7d48d7f4283b629bc40cdb449ab2022-12-21T20:34:42ZengMDPI AGPhotonics2304-67322015-09-01241027104210.3390/photonics2041027photonics2041027Assessment of Gate Width Size on Lifetime-Based Förster Resonance Energy Transfer Parameter EstimationSez-Jade Chen0Nattawut Sinsuebphon1Xavier Intes2Department of Biomedical Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY 12180, USADepartment of Biomedical Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY 12180, USADepartment of Biomedical Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY 12180, USAFörster Resonance Energy Transfer (FRET) enables the observation of interactions at the nanoscale level through the use of fluorescence optical imaging techniques. In FRET, fluorescence lifetime imaging can be used to quantify the fluorescence lifetime changes of the donor molecule, which are associated with proximity between acceptor and donor molecules. Among the FRET parameters derived from fluorescence lifetime imaging, the percentage of donor that interacts with the acceptor (in proximity) can be estimated via model-based fitting. However, estimation of the lifetime parameters can be affected by the acquisition parameters such as the temporal characteristics of the imaging system. Herein, we investigate the effect of various gate widths on the accuracy of estimation of FRET parameters with focus on the near-infrared spectral window. Experiments were performed in silico, in vitro, and in vivo with gate width sizes ranging from 300 ps to 1000 ps in intervals of 100 ps. For all cases, the FRET parameters were retrieved accurately and the imaging acquisition time was decreased three-fold. These results indicate that increasing the gate width up to 1000 ps still allows for accurate quantification of FRET interactions even in the case of short lifetimes such as those encountered with near-infrared FRET pairs.http://www.mdpi.com/2304-6732/2/4/1027Förster Resonance Energy Transfer (FRET)time-resolved imaginggate widthfluorescence lifetimegated ICCDnear infrared (NIR) dyes in vivo imaging |
| spellingShingle | Sez-Jade Chen Nattawut Sinsuebphon Xavier Intes Assessment of Gate Width Size on Lifetime-Based Förster Resonance Energy Transfer Parameter Estimation Photonics Förster Resonance Energy Transfer (FRET) time-resolved imaging gate width fluorescence lifetime gated ICCD near infrared (NIR) dyes in vivo imaging |
| title | Assessment of Gate Width Size on Lifetime-Based Förster Resonance Energy Transfer Parameter Estimation |
| title_full | Assessment of Gate Width Size on Lifetime-Based Förster Resonance Energy Transfer Parameter Estimation |
| title_fullStr | Assessment of Gate Width Size on Lifetime-Based Förster Resonance Energy Transfer Parameter Estimation |
| title_full_unstemmed | Assessment of Gate Width Size on Lifetime-Based Förster Resonance Energy Transfer Parameter Estimation |
| title_short | Assessment of Gate Width Size on Lifetime-Based Förster Resonance Energy Transfer Parameter Estimation |
| title_sort | assessment of gate width size on lifetime based forster resonance energy transfer parameter estimation |
| topic | Förster Resonance Energy Transfer (FRET) time-resolved imaging gate width fluorescence lifetime gated ICCD near infrared (NIR) dyes in vivo imaging |
| url | http://www.mdpi.com/2304-6732/2/4/1027 |
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