<https://lh5.googleusercontent.com/-N3cAlGUsoSU/T2r-_A2iTtI/AAAAAAAAAv0/5RFfB...> Sorry, the spectral mixing table was not included (see before, I failed to insert an image where I want ....) Le jeudi 22 mars 2012 11:26:38 UTC+1, jip a écrit :
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:
<https://lh3.googleusercontent.com/-dw-YEQxPyR4/T2r66g-u0EI/AAAAAAAAAvo/NLfMv...> 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<http://dip4fish.blogspot.fr/2011/05/making-mfish-image.html?showComment=1332...>. 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<https://github.com/jeanpat/MFISH>
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<http://dip4fish.blogspot.fr/2011/05/making-mfish-image.html?showComment=1332...>. 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<https://github.com/jeanpat/MFISH>
Best Regard
Jean-Patrick Pommier