MPT Image Analysis, Segmentation, Cell Counting, Cell measuring

Hello,

I’m currently working on my PhD, but I’m lost and I need your help to find the best way to measure

**- the size of the cell
**- count the nucleus and measure its size (or diameter)
**- calculate the nucleus-zytoplasm ratio

If possible automatically.

I’m working with a multiphoton microscope and a typical image looks like this (see image).

The scale of the image is set. The black spots are the nucleus and the white(ish) part surround it- is the cytoplasm of the according cell.

I found this article and tried to follow its steps “Image segmentation for integrated multiphoton microscopy and reflectance confocal microscopy imaging of human skin in vivohttp://dx.doi.org/10.3978/j.issn.2223-4292.2014.11.02
but I keep failing following the steps and don’t know why.

I would be happy if anyone could help me.
Thank you in advancel_z006-1.tif|attachment (256.4 KB)
Niklas