Multi-frequency multi-sampling fluorescence lifetime imaging using a high speed line-scan camera

Zhuang Lin; Michael Erz; Bernd Jähne

doi:10.1117/12.854763

5 May 2010 Multi-frequency multi-sampling fluorescence lifetime imaging using a high speed line-scan camera

Zhuang Lin, Michael Erz, Bernd Jähne

Proceedings Volume 7723, Optics, Photonics, and Digital Technologies for Multimedia Applications; 77231S (2010) https://doi.org/10.1117/12.854763
Event: SPIE Photonics Europe, 2010, Brussels, Belgium

Abstract

We present a theoretical multi-frequency multi-sampling frequency-domain fluorescence lifetime imaging (FLI) technique, which determines fluorescence lifetime by measuring the phase delay and decrease of the modulation of the emission relative to the excitation. Such a system could be realized with high-speed line-scan cameras using more than 100000 frames per second. It is useful for the measurement of lifetimes in the microsecond range and can be, e.g., used to measure oxygen concentrations with Ruthenium complexes. Typically 100 samples are used, resulting in a temporal resolution of microsecond fluorescence lifetime measurement. By including frequency components with frequencies much higher than the Nyquist criterion, the measurement of fluorescence lifetimes much shorter than the sampling interval is possible. In order to optimize the SNR of the predict lifetimes, we investigate the photon economy of our technique. Various ranges of the exposure time relative to the range of modulation period and compositions of multiple frequencies have been studied. The presented approach is validated by numerical simulations by Monte Carlo method.

Citation Download Citation

Zhuang Lin, Michael Erz, and Bernd Jähne "Multi-frequency multi-sampling fluorescence lifetime imaging using a high speed line-scan camera", Proc. SPIE 7723, Optics, Photonics, and Digital Technologies for Multimedia Applications, 77231S (5 May 2010); https://doi.org/10.1117/12.854763

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KEYWORDS

Modulation

Phase shift keying

Luminescence

Monte Carlo methods

Cameras

Signal to noise ratio

Fluorescence lifetime imaging

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