> Hi Curtis!!!
> Thank you for the reply!
> I tried to set the threshold without the macro, to see how reliable it is.
> There is a huge difference in intensity across the images...I really can't
> choose a good threshold for all of them. What do you mean with the
> auto-threshold algorithm?
>
>
> 2013/10/8 Curtis Rueden <
[hidden email]>
>
> > Hi Leila,
> >
> > Are you sure ImageJ isn't autoscaling your display differently across
> your
> > images?
> >
> > What does the pixel probe say? Mouse over your data and look at the raw
> > numbers spit out in the main ImageJ window's status bar.
> >
> > Also, can you use an auto-threshold algorithm across all your images to
> > make your analysis reproducible without needing to hardcode a specific
> > threshold value?
> >
> > Regards,
> > Curtis
> >
> >
> > On Tue, Oct 8, 2013 at 1:56 PM, Leila Alieh <
[hidden email]>
> wrote:
> >
> > > Hello!
> > >
> > > I developed a macro to count the number of cells positives for a given
> > > antibody. The cells are selected depending on size, circularity and
> > > intensity. I found an intensity value good for some images and I run
> the
> > > macro. After that I found that the choosen threshold is too high for
> one
> > > image, so that to have a reliable valuation for that image I should use
> > > half of the threshold chosen for the other images, but this latter
> > > threshold is really too low for the other images and it gives a wrong
> > > result. I can't understand why this image is so different because the
> > > intensity of the cells is not so different by eyes (it's a good quality
> > > image, the ab signal is strong and there are no shadows). I was
> wondering
> > > if I did some mistake during the "editing": I had to cut the images
> (I'm
> > > interested only in a small area of them) and save them in a tiff
> format.
> > Is
> > > it possible that this editing chenged the intensity values?
> > > Thank you for your help!!!!
> > >
> > > Leila
> > >
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http://imagej.nih.gov/ij/list.html> > >
> >
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>
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