Electro-optic imaging enables efficient wide-field fluorescence lifetime microscopy
Abstract
Nanosecond temporal resolution enables new methods for wide-field imaging like time-of-flight, gated detection, and fluorescence lifetime. The optical efficiency of existing approaches, however, presents challenges for low-light applications common to fluorescence microscopy and single-molecule imaging. We demonstrate the use of Pockels cells for wide-field image gating with nanosecond temporal resolution and high photon collection efficiency. Two temporal frames are obtained by combining a Pockels cell with a pair of polarizing beam-splitters. We show multi-label fluorescence lifetime imaging microscopy (FLIM), single-molecule lifetime spectroscopy, and fast single-frame FLIM at the camera frame rate with 103-105 times higher throughput than single photon counting. Finally, we demonstrate a space-to-time image multiplexer using a re-imaging optical cavity with a tilted mirror to extend the Pockels cell technique to multiple temporal frames. These methods enable nanosecond imaging with standard optical systems and sensors, opening a new temporal dimension for wide-field low-light microscopy.
- Publication:
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Nature Communications
- Pub Date:
- October 2019
- DOI:
- 10.1038/s41467-019-12535-5
- arXiv:
- arXiv:1812.06563
- Bibcode:
- 2019NatCo..10.4561B
- Keywords:
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- Physics - Optics;
- Physics - Biological Physics;
- Physics - Instrumentation and Detectors
- E-Print:
- 11 pages, 6 figures