Hi! We've got some image of gel with fluorescent immune staining. However, much of off-target fluorescence get trapped in the hydrogel, making some area very noisy. We tried to use rolling ball to substrate the background in ImageJ. However, the quality of the image is still very poor. In attachment there is one of the image (red)
Could you tell me how to improve the image (removing the bright stripe on the right)? Thank you!
You've saturated the image which is why you have that bright white stripe. It looks like its brightest because it's the most in focus spot, if you moved up and down in z I think you'd expect to see different areas become blindingly bright.
Use a lower detector gain so the image isn't saturated and compare to a control image where you haven't stained for anything to see how much of a contribution that makes.
As stated above, the bright stripe on the right depicts over-saturated areas. In simpler words, the area is pitch white and the information of the specimen is lost (i.e., unrecoverable). It is always nice to remedy -if possible- the problem from the root (i.e., the acquisition step) rather than trying to post-process the output.
Thank you! Yeah we later find a way to higher the primary antibody but lower the secondary antibody.
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