Fluorescence characteristics of human breast tissues are investigated through wavelet transform and principal component analysis (PCA). Wavelet transform of polarized fluorescence spectra of human breast tissues is found to localize spectral features that can reliably differentiate different tissue types. The emission range in the visible wavelength regime of 500–700 nm is analysed, with the excitation wavelength at 488 nm using laser as an excitation source, where flavin and porphyrin are some of the active fluorophores. A number of global and local parameters from principal component analysis of both high- and low-pass coefficients extracted in the wavelet domain, capturing spectral variations and subtle changes in the diseased tissues are clearly identifiable.