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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 23073-23085 |
| Number of pages | 13 |
| Journal | Optics Express |
| Volume | 22 |
| Issue number | 19 |
| DOIs | |
| State | Published - Sep 22 2014 |
ASJC Scopus subject areas
- Atomic and Molecular Physics, and Optics