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

Author

Listed:
  • Adam J. Bowman

    (Stanford University)

  • Brannon B. Klopfer

    (Stanford University)

  • Thomas Juffmann

    (University of Vienna
    University of Vienna)

  • Mark A. Kasevich

    (Stanford University)

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.

Suggested Citation

  • Adam J. Bowman & Brannon B. Klopfer & Thomas Juffmann & Mark A. Kasevich, 2019. "Electro-optic imaging enables efficient wide-field fluorescence lifetime microscopy," Nature Communications, Nature, vol. 10(1), pages 1-8, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-12535-5
    DOI: 10.1038/s41467-019-12535-5
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