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

12 Citations (Scopus)


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 languageEnglish
Article number288056
JournalBiomedical Optics Express
Issue number7
Pages (from-to)3132-3142
Number of pages11
Publication statusPublished - 01.07.2017

Research Areas and Centers

  • Academic Focus: Biomedical Engineering


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