I am using FIJI to get the intensity values of my confocal images, but i get different mean gray value for each time. So, I could not trust those values.
I do it like that:
1) I open the image and convert it to 8-bit image.
2) I set scale of the image.
3) From Image, I go to HiLo.
4) From Process, I substract the background, and background becomes blue. I choose Sliding Paraboloid and Disable Smoothing and preview. Rolling ball radius is 50.
5) I go to Analyze, then measure. I get the value. However, I get different values although the everything is the same. Why this is happening?
Thanks in advance.
Tugba
It's because you are changing the scale of the image, I presume manually. Make your measurements on the raw, unscaled data. You can change the brightness with the level sliders to change the display and make it brighter to see, but do not click "apply" which will then change the pixel values. I'd also leave the bit depth unchanged for measurements. You can save a separate copy that is scaled to 8 bit if you need to make images for presentation.
Thank you very much. After discussing with my colleague, I understand that I should choose the image first with ctrl+A, and then get a measurement (ctrl+M). It seems to be working now.
- Badges
- Science topic
- Similar topics
- Computer Science
- Images