Spectro-temporal encoded multiphoton microscopy and fluorescence lifetime imaging at kilohertz frame-rates

Sebastian Karpf (), Carson T. Riche, Dino Carlo, Anubhuti Goel, William A. Zeiger, Anand Suresh, Carlos Portera-Cailliau and Bahram Jalali
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Sebastian Karpf: University of California, Los Angeles (UCLA)
Carson T. Riche: University of California, Los Angeles (UCLA)
Dino Carlo: University of California, Los Angeles (UCLA)
Anubhuti Goel: University of California, Los Angeles (UCLA)
William A. Zeiger: University of California, Los Angeles (UCLA)
Anand Suresh: University of California, Los Angeles (UCLA)
Carlos Portera-Cailliau: University of California, Los Angeles (UCLA)
Bahram Jalali: University of California, Los Angeles (UCLA)

Nature Communications, 2020, vol. 11, issue 1, 1-9

Abstract: Abstract Two-Photon Microscopy has become an invaluable tool for biological and medical research, providing high sensitivity, molecular specificity, inherent three-dimensional sub-cellular resolution and deep tissue penetration. In terms of imaging speeds, however, mechanical scanners still limit the acquisition rates to typically 10–100 frames per second. Here we present a high-speed non-linear microscope achieving kilohertz frame rates by employing pulse-modulated, rapidly wavelength-swept lasers and inertia-free beam steering through angular dispersion. In combination with a high bandwidth, single-photon sensitive detector, this enables recording of fluorescent lifetimes at speeds of 88 million pixels per second. We show high resolution, multi-modal - two-photon fluorescence and fluorescence lifetime (FLIM) – microscopy and imaging flow cytometry with a digitally reconfigurable laser, imaging system and data acquisition system. These high speeds should enable high-speed and high-throughput image-assisted cell sorting.

Date: 2020
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DOI: 10.1038/s41467-020-15618-w

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