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

Sebastian Karpf*, Carson T. Riche, Dino Di Carlo, Anubhuti Goel, William A. Zeiger, Anand Suresh, Carlos Portera-Cailliau, Bahram Jalali

*Corresponding author for this work
2 Citations (Scopus)


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.

Original languageEnglish
Article number2062
JournalNature Communications
Issue number1
Publication statusPublished - 01.12.2020

Research Areas and Centers

  • Academic Focus: Biomedical Engineering


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