Multispectral imaging analysis: Spectral deconvolution and applications in biology

Multispectral imaging has been in use for over half a century. Owing to advances in digital photographic technology, multispectral imaging is now used in settings ranging from clinical medicine to industrial quality control. Our efforts focus on the use of multispectral imaging coupled with spectral deconvolution for measurement of endogenous tissue fluorophores and for animal tissue analysis by multispectral fluorescence, absorbance, and reflectance data. Multispectral reflectance and fluorescence images may be useful in evaluation of pathology in histological samples. For example., current hematoxylin/eosin diagnosis limits spectral analysis to shades of red and blue/grey. It is possible to extract much more information using multispectral techniques. To collect this information, a series of filters or a device such as an acousto-optical tunable filter (AOTF) or liquid-crystal filter (LCF) can be used with a CCD camera, enabling collection of images at many more wavelengths than is possible with a simple filter wheel. In multispectral data processing the "unmixing" of reflectance or fluorescence data and analysis and the classification based upon these spectra is required for any classification. In addition to multispectral techniques, extraction of topological information may be possible by reflectance deconvolution or multiple-angle imaging, which could aid in accurate diagnosis of skin lesions or isolation of specific biological components in tissue. The goal of these studies is to develop spectral signatures that will provide us with specific and verifiable tissue structure/function information. In addition, relatively complex classification techniques must be developed so that the data are of use to the end user.