FPGA-based multi-channel fluorescence lifetime analysis of Fourier multiplexed frequency-sweeping lifetime imaging | Zendy

Ming Zhao | Zendy; Yu Li | Zendy; Leilei Peng | Zendy

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FPGA-based multi-channel fluorescence lifetime analysis of Fourier multiplexed frequency-sweeping lifetime imaging

Author(s) -

Ming Zhao,

Yu Li,

Leilei Peng

Publication year - 2014

Publication title -

optics express

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.394

H-Index - 271

ISSN - 1094-4087

DOI - 10.1364/oe.22.023073

Subject(s) - multiplexing , fluorescence lifetime imaging microscopy , optics , mcherry , field programmable gate array , channel (broadcasting) , förster resonance energy transfer , fourier transform , confocal , materials science , computer science , physics , fluorescence , computer hardware , green fluorescent protein , chemistry , telecommunications , gene , biochemistry , quantum mechanics

We report a fast non-iterative lifetime data analysis method for the Fourier multiplexed frequency-sweeping confocal FLIM (Fm-FLIM) system [Opt. Express 22, 10221 (2014)]. The new method, named R-method, allows fast multi-channel lifetime image analysis in the system's FPGA data processing board. Experimental tests proved that the performance of the R-method is equivalent to that of single-exponential iterative fitting, and its sensitivity is well suited for time-lapse FLIM-FRET imaging of live cells, for example cyclic adenosine monophosphate (cAMP) level imaging with GFP-Epac-mCherry sensors. With the R-method and its FPGA implementation, multi-channel lifetime images can now be generated in real time on the multi-channel frequency-sweeping FLIM system, and live readout of FRET sensors can be performed during time-lapse imaging.

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