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. Author manuscript; available in PMC: 2019 Jan 1.
Published in final edited form as: J Biophotonics. 2017 May 9;11(1):10.1002/jbio.201600227. doi: 10.1002/jbio.201600227

Figure 2.

Figure 2

Experimental validation of the theoretical sensitivity analysis methodology was performed by measuring Calcein Green uptake in HEK293 cells labeled with NucBlue (nuclear marker) using a Nikon A1R spectral confocal microscope. A) Transmitted light image to visualize cellular outlines. B) Spectral library used for linear unmixing. C) Merged, false-colored composite of unmixed images for 3 selected time points: 0, 1050, and 1800 seconds. NucBlue labeling is shown as blue, non-specific NucBlue labeling as red, and Calcein Green as green. Note that by the final time point, several nuclear regions exhibit a bright yellow color, indicating pixels that are oversaturated on at least one wavelength band.