title: FPGA-based multi-channel fluorescence lifetime analysis of Fourier multiplexed frequency-sweeping lifetime imaging.
abstract: 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.
authors.forename: Ming
authors.initials: M
authors.lastname: Zhao
authors.forename: Yu
authors.initials: Y
authors.lastname: Li
authors.forename: Leilei
authors.initials: L
authors.lastname: Peng
mesh_terms: D003952:Diagnostic Imaging; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D049408:Luminescent Agents; D008856:Microscopy, Fluorescence / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes; D049408:Luminescent Agents; D049452:Green Fluorescent Proteins