TY - JOUR
T1 - Single pulse two photon fluorescence lifetime imaging (SP-FLIM) with MHz pixel rate
AU - Eibl, Matthias
AU - Karpf, Sebastian
AU - Weng, Daniel
AU - Hakert, Hubertus
AU - Pfeiffer, Tom
AU - Kolb, Jan Philip
AU - Huber, Robert
PY - 2017/7/1
Y1 - 2017/7/1
N2 - 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.
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.
UR - http://www.scopus.com/inward/record.url?scp=85023642864&partnerID=8YFLogxK
U2 - 10.1364/BOE.8.003132
DO - 10.1364/BOE.8.003132
M3 - Journal articles
AN - SCOPUS:85023642864
SN - 2156-7085
VL - 8
SP - 3132
EP - 3142
JO - Biomedical Optics Express
JF - Biomedical Optics Express
IS - 7
M1 - 288056
ER -