Differentiation index and fusion index

Dear all,

I was wondering if anyone had experience and or a script for the use of Image J in the assessment of myogenic differentiation index or fusion index. Differentiation index is defined as the number of nuclei in myotubes expressing myosin heavy chain divided by the total number of nuclei in a field. The fusion index is defined as the number of myosin heavy chain expressing myotubes with greater than 2 nuclei divided by the total number of nuclei.

Thank you so much

Sincerely
Mari

Hi @MyogNCI, welcome to the forum :slight_smile:

This seems like straightforward mathematic operations, but before you get to this part you have to extract the required information from your images. I.e. you have to develop a script or macro that can

  1. detect and count the nuclei expressing myosin heavy chain
  2. detect the myotubes to determine if a nuclei is inside of or outside of the myotube

Once you have this information the math is just a couple of lines of code. Getting the information is the hard part, although it could also be easy, depending on the quality of your data.

If you could share some example images, users of the forum can suggest different ways to extract this information from the image :slight_smile:

Here is a representative image - green is MHC and blue is DAPI

Thank you so much