DAB staining

Hi all!

I’m still working on my DAB stained tissues… and I still have problems!

This time I don;t know why when I separate the channels and my DAB one is clearly empty the software recognize something…
WHY???

Thanks!


It seems that the threshold is 0, or very close to it. Either you manually set it to 0 (I can’t tell since you didn’t attach a pipeline or example picture), or was automatically determined. I take it from your question that this is DAB-negative control? If so, you may want to set the lower bound to something higher than 0, to a value sufficient to get rid of these false positives.

Regards,
-Mark

THank you for the answer!
actually I was trying to load more than this picture but something was wrong…
This is the pipeline ( you suggest me some months ago) and this is the picture. yes it is a negative control, so my DAB channel should be empty. and as you suggest if I set the lower bound to 0.2 i don’t have any reading!

Thank you!
CON1.tif.zip (6.44 MB)
DAB pipeline.cp (7 KB)

It looks like you set your threshold correction factor quite low (0.1), which is fairly unusual for assays I’ve seen. Was it necessary to set it that low for get the results you want for other images?

I ask since if you set the threshold correction factor to 1 (the default value) and the lower threshold bound to 0, no objects are identified (which is what you want, correct?).
-Mark