I need help to analyse these two files of images !
I am trying to do ratiometric measurments means( Magenta/ Red images )
The workflow is:
- Images >>> Stacks >>> Stack to images
- Images calculator then I divide 2/3 image by 3/3 image
- Substract the control mean intensity from the resulted image
- Edit >>> selection >>>creat selection of thresholded image
- Choose the original result image >>> Edit >>>> restore selection
- Analyse >>>> set measurements >> chose the result image
- Measure >>> Mean intensity
I am intresting to see the change of ratiometric measurements in response to oxygen concentration.
Each resulted image has a britghtness and contrast, is that affect the mean intensity?
Should I keep the maximum and minimum brigtness the same to get right intensity?