Single pulse two photon fluorescence lifetime imaging (SP-FLIM) with MHz pixel rate

Matthias Eibl; Sebastian Karpf; Daniel Weng; Hubertus Hakert; Tom Pfeiffer; Jan Philip Kolb; Robert Huber

doi:10.1364/BOE.8.003132

Single pulse two photon fluorescence lifetime imaging (SP-FLIM) with MHz pixel rate

Matthias Eibl, Sebastian Karpf, Daniel Weng, Hubertus Hakert, Tom Pfeiffer, Jan Philip Kolb, Robert Huber

California State University Los Angeles

36 Citations (Scopus)

Abstract

Two-photon-excited fluorescence lifetime imaging microscopy (FLIM) is a chemically specific 3-D sensing modality providing valuable information about the microstructure, composition and function of a sample. However, a more widespread application of this technique is hindered by the need for a sophisticated ultra-short pulse laser source and by speed limitations of current FLIM detection systems. To overcome these limitations, we combined a robust sub-nanosecond fiber laser as the excitation source with high analog bandwidth detection. Due to the long pulse length in our configuration, more fluorescence photons are generated per pulse, which allows us to derive the lifetime with a single excitation pulse only. In this paper, we show high quality FLIM images acquired at a pixel rate of 1 MHz. This approach is a promising candidate for an easy-to-use and benchtop FLIM system to make this technique available to a wider research community.

Original language	English
Article number	288056
Journal	Biomedical Optics Express
Volume	8
Issue number	7
Pages (from-to)	3132-3142
Number of pages	11
DOIs	https://doi.org/10.1364/BOE.8.003132
Publication status	Published - 01.07.2017

Funding

European Union project ENCOMOLE-2i (Horizon 2020, ERC CoG no. 646669); German Research Foundation (DFG project HU1006/6 and EXC 306/2). We would like to acknowledge support from A. Vogel at the Institute of Biomedical Optics (BMO) at the University of L?beck. Moreover we would like to thank Gereon H?ttmann (BMO) for helpful discussions on multi-photon imaging technology and Astrid Link and Ramtin Rahmanzadeh (both BMO), Christin Lehmann (Institute for anatomy), and Zouhair Aherrahrou (Institute for Integrative and Experimental Genomics) for fruitful discussions on cell imaging and help with sample preparation.

UN SDGs

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

SDG 9 Industry, Innovation, and Infrastructure

Research Areas and Centers

Academic Focus: Biomedical Engineering

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10.1364/BOE.8.003132

Cite this

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abstract = "Two-photon-excited fluorescence lifetime imaging microscopy (FLIM) is a chemically specific 3-D sensing modality providing valuable information about the microstructure, composition and function of a sample. However, a more widespread application of this technique is hindered by the need for a sophisticated ultra-short pulse laser source and by speed limitations of current FLIM detection systems. To overcome these limitations, we combined a robust sub-nanosecond fiber laser as the excitation source with high analog bandwidth detection. Due to the long pulse length in our configuration, more fluorescence photons are generated per pulse, which allows us to derive the lifetime with a single excitation pulse only. In this paper, we show high quality FLIM images acquired at a pixel rate of 1 MHz. This approach is a promising candidate for an easy-to-use and benchtop FLIM system to make this technique available to a wider research community.",

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AB - Two-photon-excited fluorescence lifetime imaging microscopy (FLIM) is a chemically specific 3-D sensing modality providing valuable information about the microstructure, composition and function of a sample. However, a more widespread application of this technique is hindered by the need for a sophisticated ultra-short pulse laser source and by speed limitations of current FLIM detection systems. To overcome these limitations, we combined a robust sub-nanosecond fiber laser as the excitation source with high analog bandwidth detection. Due to the long pulse length in our configuration, more fluorescence photons are generated per pulse, which allows us to derive the lifetime with a single excitation pulse only. In this paper, we show high quality FLIM images acquired at a pixel rate of 1 MHz. This approach is a promising candidate for an easy-to-use and benchtop FLIM system to make this technique available to a wider research community.

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Single pulse two photon fluorescence lifetime imaging (SP-FLIM) with MHz pixel rate

Abstract

Funding

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