Fig. 3: Full spectral fluorescence lifetime imaging of a fixed honeybee wing.
From: Full spectrum fluorescence lifetime imaging with 0.5 nm spectral and 50 ps temporal resolution
a Intensity image summed from the whole FS-FLIM data cube, a color image of the intensity data produced as per Fig. 2 showing normalized spectral variations across the sample along with normalized spectra from two regions of interest (ROI) within the sample highlighting the subtle spectral variations. b Slices through the FS-FLIM cube at eight wavelengths (top), the emission spectrum for the whole sample colored by emission wavelength, and the lifetime histograms for each presented wavelength. An exposure time of 500 µs per pixel was used to provide sufficient signal to noise ratio for the majority of the emission spectrum. Where the signal was reduced at the wavelength limits of the spectrum a moving spectral averaging filter was applied of a width of 8 pixels (~4 nm) to insure an adequate signal to noise ratio of >10. Whilst the use of such a moving filter does impact on the overall spectral resolution on these areas, far more information is retained than using static spectral binning as is enforced by systems with fewer spectral collection channels. The sample was imaged 3 times to insure repeatability.
