Each image is a raw image of a given fluorochrome.

A given fluorochrome targets a subset of chromosomes, more informations can be found in:

Automatic Segmentation and Classification of
Multiplex-Fluorescence In-Situ Hybridization
Chromosome Images
by
Hyo Hun Choi, B.S., M.S.

Choi gives an example of CCD integration times:

An example of integration times is [DAPI, Aqua,
Green, Gold, Red, Far Red] = [0.14, 6, 0.76, 6, 2.96, 1.4] seconds

He also gives an example of spectral mixing in the raw images:

With the vysis data set (example:https://github.com/jeanpat/MFISH/tree/master/MFISH_TIFF/V13__40), a given fluorochrome corresponds to several chromosomes, in the spectrumAqua  image the chromosomes 3,10,11,13,15,20,21,22,X,Y are visibles.

A: S. Aqua

D DAPI

F: Far Red

G: S. Green

R: S. Red

Y: S. Gold

To perform "perfect" spectral unmixing, I suppose it's necessary to know the integration time, the characteristic of the microscope bandpass filters, the emission spectrum of each fluorochrome; but would it be possible to make some kind of "estimated" unmixing from only the images themselves?

Best regards
Jean-Pat

Le mercredi 21 mars 2012 12:10:16 UTC+1, jip a écrit :

Dear all,
Some one asked me how I choose the matrix coef to convert a five multispectral image to a three (RGB) multispectral image. I feel that there is a best way to do that by maximising some distance in a color space.
How would you do that?
Examples of multispectral images are available here

Best Regard

Jean-Patrick Pommier

Le mercredi 21 mars 2012 12:10:16 UTC+1, jip a écrit :

Dear all,
Some one asked me how I choose the matrix coef to convert a five multispectral image to a three (RGB) multispectral image. I feel that there is a best way to do that by maximising some distance in a color space.
How would you do that?
Examples of multispectral images are available here

Best Regard

Jean-Patrick Pommier