Thresholding hematoxylin-stained nuclei

Posted by nwoller on Jan 12, 2011; 3:36pm
URL: http://imagej.273.s1.nabble.com/Thresholding-hematoxylin-stained-nuclei-tp3686020.html

Hi,

I am a new ImageJ user. I have taken a series of images of a slide with tonsil tissue which has been stained with hematoxylin as well as liquid permanent red (Immunostain). I have been careful to capture all of the images in exactly the same way, same light intensity, same exposure time, same white balance etc. I would like to count the number of hematoxylin-stained nuclei in my images. I threshold the image, apply a watershed and use "analyze particles". Unfortunately, I don't seem to be able to apply the same thresholds to my images. A threshold that works nicely for one image is completely hopeless for another image. It is possible that the intensity of the hematoxylin stain varies quite alot in the same tissue. A colleague therefore suggested that I should threshold by finding the top of the "threshold curve" and using that value for thresholding my images. What do you think? I would really appreciate some help and advice.

Nina