A Method for Paired Comparisons of Glo Germ Quantity in Images of Hands Before and After Washing.

We present a reproducible pipeline that converts color images into quantitative fluorescence maps by combining spectral measurement with a linear mixture model. The method is designed specifically for quantitative comparisons of Glo Germ™ on images of hands taken under different experimental conditions with controlled illumination. The emission spectrum of Glo Germ is measured using a spectral photometer and normalized to obtain its spectral power density function. This spectrum is projected into CIE XYZ coordinates and incorporated into a linear mixture model in which each pixel contains contributions from white light, UV-illuminated skin reflectance, and fluorophore emission. Component magnitudes are estimated with non-negative least squares, yielding a grayscale image whose intensity is a monotonic proxy for local fluorophore density. Spatial integration provides an image-level summary proportional to total detected material. Compared with single-channel proxies, the observer suppresses background structure, improves contrast, and remains radiometrically interpretable. Because the method depends only on measurable spectra and linear transforms, it can be reproduced across cameras and extended to other fluorophores.