A novel analytical spectral-imaging system and its results in the examination of biological specimens are presented. The SpectraCube 1000 system measures the transmission, absorbance, or fluorescence spectra of images studied by light microscopy. The system is based on an interferometer combined with a CCD camera, enabling measurement of the interferogram for each pixel constructing the image. Fourier transformation of the interferograms derives pixel by pixel spectra for 170 X 170 pixels of the image. A special `similarity mapping' program has been developed, enabling comparisons of spectral algorithms of all the spatial and spectral information measured by the system in the image. By comparing the spectrum of each pixel in the specimen with a selected reference spectrum (similarity mapping), there is a depiction of the spatial distribution of macromolecules possessing the characteristics of the reference spectrum. The system has been applied to analyses of bone marrow blood cells as well as fluorescent specimens, and has revealed information which could not be unveiled by other techniques. Similarity mapping has enabled visualization of fine details of chromatin packing in the nucleus of cells and other cytoplasmic compartments. Fluorescence analysis by the system has enabled the determination of porphyrin concentrations and distribution in cytoplasmic organelles of living cells.

Date Published: 1 February 1995
PDF: 5 pages
Proc. SPIE 2329, Optical and Imaging Techniques in Biomedicine, (1 February 1995); doi: 10.1117/12.200882

Published in SPIE Proceedings Vol. 2329:
Optical and Imaging Techniques in Biomedicine
Hans-Jochen Foth; Aaron Lewis; Halina Podbielska M.D.; Michel Robert-Nicoud; Herbert Schneckenburger; Anthony J. Wilson, Editor(s)