Martin De Biasio, Thomas Arnold, Raimund Leitner, and Franz G. Würtz
M-FISH, spectral unmixing, ALS, VARIMAX
M-FISH images are difficult to interpret because the emission spectra of fluorochrome marked DNA probes overlap with each other and with the tissue's intrinsic auto-fluorescence. Multi-spectral imaging is used to take spatially and spectrally resolved images of M-FISH samples and spectral unmixing is applied to resolve the crosstalk in the emission spectra. The unmixing is realized by a combination of: Alternating Least Squares (ALS) and VARIMAX. Spot counting algorithms use the high contrast false color images to automatically determine the HER-2/neu to CEP 17 ratio.
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