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fluorescence quantification and normalization in tissue slides

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fluorescence quantification and normalization in tissue slides

Davide.Priori
3 posts
Hi ! I'm new in this forum because I'm starting to use ImageJ just now but I have already some problems and I need of your professional help to solve them. I explain you briefly : my aim is to quantify the fuorescence intensity in small intestinal tissue slides stained with fluo-antybody in order to compare the fluorescence from 50 different microscope slides.
I have some problem to do it:
1) I want to get images in RGB filters. But my staining will be only in green (antibody) and blue (nuclei). What is the best image format ? (.tiff, .zvi, .jpeg..?)
2) i think to set the same threshold for all images I get.
3) I think to normalize the fluorescence intensity and area risults with DAPI area in order to obtain value relative to an amount tissue value.
4) I think to do it in a select area (ROI) in the center of images in order to avoid unwanted regions and because I think in the center the intensity images is better define.

I don't know how to do it ! in particular I don't know how can I set the same conditions for all the images-
Do you have any suggestion about pugl-in and macros to use in this case??

Thank you all !!

 Davide

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