SPECTRAL IMAGING FOR QUANTITATIVE HISTOLOGY AND CYTOGENETICS

Evaluation of cell morphology by bright field microscopy is the pillar of histopathological diagnosis. The need for quantitative and objecti ve parameters for diagnosis gave rise to the development of morphometr ic methods, Morphometry combined with spectral imaging provides multi- pixel information from a specimen, which can be used for further image processing and quantitative analysis. The spectroscopic analysis is b ased on the ability of a stained histological specimen to absorb, refl ect, or emit photons in ways characteristic to its interactions with s pecific dyes. Spectral information obtained from a histological specim en is stored in a cube whose appellate signifies the two spatial dimen sions of a flat sample (x and y) and the third dimension, the spectrum , representing the light intensity for every wavelength. By mathematic al analysis of the cube database, it is possible to perform the functi on of spectral-similarity mapping (SSM) which enables the demarcation of areas occupied by the same type of material. Spectral similarity ma pping constructs new images of the specimen, revealing areas with simi lar stain-macromolecule characteristics and enhancing subcellular feat ures. Spectral imaging combined with SSM reveals nuclear organization and identifies specifically the nucleoli domains. Therefore, different iation stages as well as apoptotic and necrotic conditions are easily quantified. The commercial SpectraCube(TM) system was developed for th e application of spectral imaging in biology, recording both transmitt ed light and fluorescence. The SKY technique utilizes the advantages o f the SpectraCube(TM) for multi probe FISH and chromosome karyotyping, identifying marker chromosomes, detecting subtle chromosome transloca tions and clarifying complex karyotypes.