ImagePro>Ratiometric measurements


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ImagePro>Ratiometric measurements



Hi!
 
I am doing pH measurements in cells with the ratiometric probe BCECF. Could somebody tell me what is the usual procedure to obtain good ratio images from images at two different wavelengths ? I did the following:
1) Extract and subtrack background from the two images
2) register the two images
3) convert into floating point images
4) divide image 1 by image 2
 
The resulting image is very noisy, due to low pixel values in image 2 (in particular where there are no cells). How can I improve this procedure ?
 
Thanks for your help
 
Christian Brière
UMR CNRS/UPS 5546
24 chemin de borderouge
BP17 Auzeville 31326 Castanet-tolosan (France)
Tel: 05 62 19 35 90
Fax: 05 62 19 35 02



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