Multispectral auto-fluorescence imaging is a novel technique that leverages the tissues' and microbiome's native fluorescence and aids in label free detection. Biomarkers like NADH, Flavin, Porphyrin, Di-tyrosine linkages, Chitin, collagen etc. have characteristic emission when excited with various wavelength of light sources. Analysis of these spectral images at various emission bands and applying machine learning/deep learning techniques on these images can be effectively used in label free detection from pathogens to antibiotic susceptibility testing to cancer diagnostics.