By taking advantage of the continuous spectrum collected for each image pixel by the Hyperspectral Microscopy Imaging (HMI) system, the data spectra can be de-convolved into a set of standard spectra coefficients for each pixel. The coefficients are calculated by the analysis program and indicate the relative amounts of each standard necessary to reproduce the data spectra for each image pixel. Images of a single color or composite images of two or more colors can be produced for visual analysis. The HMI system is composed of an Olympus inverted microscope, imaging spectrograph, CCD camera, motorized X-Y stage and illumination sources. This system has been used to scan and analyze pathology tissue samples which have been stained with 4 standard fluorochromes (not including the dynamic background spectrum) attached to specific antibodies. The typical wavelength range is 420nm to 785nm with the longest wavelength markers emitting in the spectral region where the human eye is not sensitive. Quantitative values are recorded and can be compared to the visual interpretations.

Date Published: 12 February 2009
PDF: 10 pages
Proc. SPIE 7182, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII, 71821F (12 February 2009); doi: 10.1117/12.809277

Published in SPIE Proceedings Vol. 7182:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)