Quantify fluorescence in cell surface marker of hepatocytes

Hi,

I have a set of images of hepatocytes with a primary antibody used against a cell surface marker. I’m trying to quantify the change in fluorescence between wildtype and my samples where this surface marker has been repressed. Is there a recommended pipeline I should follow? More specifically, I’m thinking of subtracting my nuclei from cells and quantifying the fluoresence of the cells. Can this be achieved?

Hi,
This can definitely be done! Look at this forum thread and this example for pipelines that do nuclear-to-cytoplasm measurements- you should just be able to remove some unnecessary modules and fiddle with some settings to adapt them for your purposes.
Good luck!

Thank you! Would this work even though my protein resides on the cell wall and not in the cytoplasm? I’ve attached some images, with nuclei stained with DAPI and with my protein of interest stained with GFP. Thanks! Uploading…
Uploading…

It seems likely, though if you want to measure the cell wall and only the cell wall you may want to try slightly different approaches- I found this post from just searching the forums for “membrane quantification” and there may be others of interest as well. Tinker with it and let us know if you have specific questions.
Good luck!