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. Author manuscript; available in PMC: 2013 Jan 1.
Published in final edited form as: J Biophotonics. 2011 Oct 11;5(1):67–84. doi: 10.1002/jbio.201100066

Figure 3.

Figure 3

Theoretical sensitivity analysis revealed that single-band fluorescence imaging assays can be designed to have either high sensitivity with low specificity or low sensitivity with slightly improved specificity. A 30 × 30 pixel square of GFP signal was added to control hyperspectral images (A). Wavelengths between 500-550 nm were summed to approximate a standard GFP emission filter and thresholded to detect GFP signal based on intensity differences (B). Thresholded data are shown without error bars (C) to visualize the dependence of sensitivity (the minimum amount of GFP added that can be detected) and specificity (type I errors are represented by the number of GFP-positive pixels when 0 GFP was added); and with error bars (± SEM, n = 16 fields-of-view) to illustrate the high variance that is inherent in using single-band thresholding for GFP detection when there is a high level of autofluorescence (D).