2025-08-05 スイス連邦工科大学ローザンヌ校（EPFL）
＜関連情報＞
- https://actu.epfl.ch/news/new-method-instantly-characterizes-thousands-of-mo/
- https://www.nature.com/articles/s41377-025-01901-2
ゲート付き単一光子カメラを用いた単一分子広視野蛍光寿命イメージング Wide-field fluorescence lifetime imaging of single molecules with a gated single-photon camera
Nathan Ronceray,Salim Bennani,Marianna Fanouria Mitsioni,Nicole Siegel,Maria J. Marcaida,Claudio Bruschini,Edoardo Charbon,Rahul Roy,Matteo Dal Peraro,Guillermo P. Acuna & Aleksandra Radenovic
Light: Science & Applications Published:04 August 2025
DOI:https://doi.org/10.1038/s41377-025-01901-2
Abstract
Fluorescence lifetime imaging microscopy (FLIM) is a powerful tool to discriminate fluorescent molecules or probe their nanoscale environment. Traditionally, FLIM uses time-correlated single-photon counting (TCSPC), which is precise but intrinsically low-throughput due to its dependence on point detectors. Although time-gated cameras have demonstrated the potential for high-throughput FLIM in bright samples with dense labeling, their use in single-molecule microscopy has not been explored extensively. Here, we report fast and accurate single-molecule FLIM with a commercial time-gated single-photon camera. Our optimized acquisition scheme achieves single-molecule lifetime measurements with a precision only about three times less than TCSPC, while imaging with a large number of pixels (512 × 512) allowing for the spatial multiplexing of over 3000 molecules. With this approach, we demonstrate parallelized lifetime measurements of large numbers of labeled pore-forming proteins on supported lipid bilayers, and temporal single-molecule Förster resonance energy transfer measurements at 5-25 Hz. This method holds considerable promise for the advancement of multi-target single-molecule localization microscopy and biopolymer sequencing.
