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

doi:10.1038/s41467-020-15618-w

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

Institute of Biomedical Optics

California State University Los Angeles

70 Citations (Scopus)

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.

Original language	English
Article number	2062
Journal	Nature Communications
Volume	11
Issue number	1
ISSN	1751-8628
DOIs	https://doi.org/10.1038/s41467-020-15618-w
Publication status	Published - 01.12.2020

Funding

This research was sponsored in part by the National Institutes of Health grants 5R21GM107924-03 to B.J. and C.P.-C. and R21EB019645 to B.J., Cal-BRAIN grant 350050 (California Blueprint for Research to Advance Innovations in Neuroscience) to B.J. and C.P.-C., as well as grant W81XWH-14-1-0433 (USAMRMC, DOD), and NIH NICHD grant R01 HD054453 to C.P.-C. and by ImPACT Program of the Council of Science, Technology and Innovation (Cabinet office, Government of Japan) to C.R. and D.D.C. Sebastian Karpf gratefully acknowledges a postdoctoral research fellowship from the German Research Foundation (DFG, project KA 4354/1-1), the Juniorprofessorship with financial support by the state of Schleswig-Holstein (Excellence chair program by the universities Kiel and Luebeck) and funding from the Deutsche For-schungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy – EXC 2167-390884018.

Research Areas and Centers

Academic Focus: Biomedical Engineering

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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title = "Spectro-temporal encoded multiphoton microscopy and fluorescence lifetime imaging at kilohertz frame-rates",

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.",

author = "Sebastian Karpf and Riche, \{Carson T.\} and \{Di Carlo\}, Dino and Anubhuti Goel and Zeiger, \{William A.\} and Anand Suresh and Carlos Portera-Cailliau and Bahram Jalali",

note = "Funding Information: This research was sponsored in part by the National Institutes of Health grants 5R21GM107924-03 to B.J. and C.P.-C. and R21EB019645 to B.J., Cal-BRAIN grant 350050 (California Blueprint for Research to Advance Innovations in Neuroscience) to B.J. and C.P.-C., as well as grant W81XWH-14-1-0433 (USAMRMC, DOD), and NIH NICHD grant R01 HD054453 to C.P.-C. and by ImPACT Program of the Council of Science, Technology and Innovation (Cabinet office, Government of Japan) to C.R. and D.D.C. Sebastian Karpf gratefully acknowledges a postdoctoral research fellowship from the German Research Foundation (DFG, project KA 4354/1-1), the Juniorprofessorship with financial support by the state of Schleswig-Holstein (Excellence chair program by the universities Kiel and Luebeck) and funding from the Deutsche For-schungsgemeinschaft (DFG, German Research Foundation) under Germany{\textquoteright}s Excellence Strategy – EXC 2167-390884018. Publisher Copyright: {\textcopyright} 2020, The Author(s). Copyright: Copyright 2020 Elsevier B.V., All rights reserved.",

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doi = "10.1038/s41467-020-15618-w",

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AU - Riche, Carson T.

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AU - Goel, Anubhuti

AU - Zeiger, William A.

AU - Suresh, Anand

AU - Portera-Cailliau, Carlos

AU - Jalali, Bahram

N1 - Funding Information: This research was sponsored in part by the National Institutes of Health grants 5R21GM107924-03 to B.J. and C.P.-C. and R21EB019645 to B.J., Cal-BRAIN grant 350050 (California Blueprint for Research to Advance Innovations in Neuroscience) to B.J. and C.P.-C., as well as grant W81XWH-14-1-0433 (USAMRMC, DOD), and NIH NICHD grant R01 HD054453 to C.P.-C. and by ImPACT Program of the Council of Science, Technology and Innovation (Cabinet office, Government of Japan) to C.R. and D.D.C. Sebastian Karpf gratefully acknowledges a postdoctoral research fellowship from the German Research Foundation (DFG, project KA 4354/1-1), the Juniorprofessorship with financial support by the state of Schleswig-Holstein (Excellence chair program by the universities Kiel and Luebeck) and funding from the Deutsche For-schungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy – EXC 2167-390884018. Publisher Copyright: © 2020, The Author(s). Copyright: Copyright 2020 Elsevier B.V., All rights reserved.

PY - 2020/12/1

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N2 - 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.

AB - 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.

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ER -

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

Abstract

Funding

Research Areas and Centers

UN SDGs

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