Electro-optic imaging enables efficient wide-field fluorescence lifetime microscopy

Adam J. Bowman (), Brannon B. Klopfer, Thomas Juffmann and Mark A. Kasevich
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Adam J. Bowman: Stanford University
Brannon B. Klopfer: Stanford University
Thomas Juffmann: University of Vienna
Mark A. Kasevich: Stanford University

Nature Communications, 2019, vol. 10, issue 1, 1-8

Abstract: 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.

Date: 2019
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DOI: 10.1038/s41467-019-12535-5

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