ImageJ overexposes images

imagej

#1

I have a problem when working with Z stacks in ImageJ. I acquire the image on Olympus microscope and open the.vsi image via Olympus plug-in or .tif image via regular ImageJ option. However, in both cases ImageJ overexposes certain channels that are actually not overexposed in the original. Does somebody know how to fix this?


#2

Good day,

I assume that your stacks contain images that are 16bit. To correctly display 16bit images you need to go to the “Image >> Adjust >> Brightness/Contrast…”-dialog and click Reset.

This action doesn’t change your images but only their display on the computer monitor. This is necessary because there are no monitors that can dispaly 16bit image. Therefore and for display only, the enormous 16bit range is “compressed”.

Regards

Herbie


#3

If I do that, can I still quantify fluorescence intensity or after the image manipulation it is not correct anymore?

Best regards,

Ana.


#4

Ana,

please read thoroughly:

This action doesn’t change your images but only their display on the computer monitor.

Consequently and not only regarding

can I still quantify fluorescence intensity

the answer is yes!

As always if you encounter problems or have questions, please study the User Guide
https://imagej.nih.gov/ij/docs/guide/index.html
and in your case especially this section:
https://imagej.nih.gov/ij/docs/guide/146-28.html#toc-Subsection-28.2

Regards

Herbie


#5

Thank you! :slight_smile:

Best regards,
Ana.