Fluorescence spectral data acquired from tissues in vivo or in vitro is
processed in accordance with a multivariate statistical method to achieve
the ability to probabilistically classify tissue in a diagnostically
useful manner, such as by histopathological classification. The apparatus
includes a controllable illumination device for emitting electromagnetic
radiation selected to cause tissue to produce a fluorescence intensity
spectrum. Also included are an optical system for applying the plurality
of radiation wavelengths to a tissue sample, and a fluorescence intensity
spectrum detecting device for detecting an intensity of fluorescence
spectra emitted by the sample as a result of illumination by the
controllable illumination device. The system also include a data
processor, connected to the detecting device, for analyzing detected
fluorescence spectra to calculate a probability that the sample belongs
in a particular classification. The data processor analyzes the detected
fluorescence spectra using a multivariate statistical method. The five
primary steps involved in the multivariate statistical method are (i)
preprocessing of spectral data from each patient to account for
inter-patient variation, (ii) partitioning of the preprocessed spectral
data from all patients into calibration and prediction sets, (iii)
dimension reduction of the preprocessed spectra in the calibration set
using principal component analysis, (iv) selection of the diagnostically
most useful principal components using a two-sided unpaired student's
t-test and (v) development of an optimal classification scheme based on
logistic discrimination using the diagnostically useful principal
component scores of the calibration set as inputs.