Open AccessFPGA-based multi-channel fluorescence lifetime analysis of Fourier multiplexed frequency-sweeping lifetime imaging
Author(s) -
Ming Zhao,
Yu Liu,
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.