How to normalize signal?

Hello all!

This is my first time posting on here, but I am a bit stumped as to how I should do this. I am trying to do quantification of the protocerebral bridge, using Dapi and GFP at 2 micrometers thick slices. The problem is that I am trying to normalize the GFP to the Dapi, and I have not done this before. I have MaxStacks created, but since I have not done this before, I am trying to figure out how to do this. Can anyone help me? I’d very much appreciate it.

Thank you!

Hey @JDSL1998,

could you provide some example data? Furthermore, it would be interesting why do want to normalize the image? Last but not least: What do you want to measure? What’s the target physical unit? Some intensity or area or volume?

Cheers,
Robert

Thank you for responding, Robert!

MAX_Composite (RGB) QUAS QA 2 03062020.tif (3.0 MB) MAX_Composite (RGB) QUAS QA 18 DEG BRAIN 4 03162020.tif (3.0 MB)

My hope is these two images will load. I have two MaxStack images I created here with my control and my experimental. My PI would like for me to normalize these images because the way our Dapi looked so often in these images was odd to her. It oftentimes looked as if it was bleeding through. As a result, she would like for me to normalize based on slices I make of the protocerebral bridge, accounting for the area and the intensity in particular. I am just unsure how to move from there, but am trying to get this figured out as soon as possible. Thank you very much again!

Best,
Jacob