How to quantify microtubule filaments in my irregular-shaped cells

Hello everyone,

I’m trying to find a way to quantify the amount of microtubule filaments there are in my microscopy pictures but am running into a bit of trouble.

After looking through many forums, I decided that the best way to quantify the filaments per cell would be to use this method:

Basically, I would take my images, run them through a difference of gaussian and follow that with ridge detection.

My problem is that I currently don’t have a way to segment my cells automatically.
I could free-hand segment them, but this would not be very replicable or precise.

Is there a way to separate my cells so I can calculate the amount of filaments per cell (density)?
I’m also open to suggestions on alternative ways to go about quantifying the amount of microtubules per cell.
My end goal is to compare the amount of filaments in each cell from one treatment group to another.

I have attached a picture of what I’m working with below.

example.tiff (943.5 KB)