Highly Multiplexed Confocal Fluorescence Lifetime Microscope Designed for Screening Applications
| Autor: | Nehad Hirmiz, Elizabeth J Osterlund, Morgan Richards, David W. Andrews, Qiyin Fang, Anthony Tsikouras |
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| Rok vydání: | 2021 |
| Předmět: |
Fluorescence-lifetime imaging microscopy
Materials science Microscope business.industry Confocal Avalanche photodiode Fluorescence Small molecule Atomic and Molecular Physics and Optics law.invention Förster resonance energy transfer law Microscopy Optoelectronics Electrical and Electronic Engineering business |
| Zdroj: | IEEE Journal of Selected Topics in Quantum Electronics. 27:1-9 |
| ISSN: | 1558-4542 1077-260X |
| DOI: | 10.1109/jstqe.2020.2997834 |
| Popis: | Protein-protein interactions can be measured in live cells, at nanometer scale, using Fluorescence Lifetime Imaging Microscopy (FLIM) enabled Forster Resonance Energy Transfer (FRET). There are growing interests in exploring protein-protein interactions in drug discovery applications. Traditional single point confocal microscopes, however, are slow and unsuited to small molecule screening, especially when combined with FLIM-FRET. We developed a 32 × 32 multiplexed confocal microscope, which employs a single-photon avalanche photodiode array with time gating capabilities for rapid FLIM acquisition. It has been demonstrated that such multiplexing technique can capture a 960 × 960 pixel multi-channel confocal fluorescence lifetime images in less than 1.5 seconds. Binding curves of two Bcl-2 family proteins: Bcl-XL and Bad were generated in live cells imaging experiments. The results show that the small molecule inhibitor A-1131852 is a more effective compound for disrupting Bcl-XL binding to Bad than ABT-263, which demonstrated the feasibility of screening of protein-protein interactions in high density well-plates. |
| Databáze: | OpenAIRE |
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