Help with Data Analysis

[Lacey Elwyn]

Dear ImageJ Experts,

I have attached a sample image of the innervated artery for which I am interested in analyzing. I would like to use ImageJ to quantify the density of nerves in a selected area of the artery. I will inform you of what I have considered to accomplish this and look forward to your advise. I am not sure if I am even on the right tract.

I have specified a standard box size (Edit, Selection, Specify, Box Size, 200x200, for example) to place on the artery. I have considered thresholding the image. I could pick an average pixel value for the background and an average pixel value for the nerve fibers and have the program only select the nerve fibers from the photo in order to quantify density of innervation. However, I do not know how to determine the pixels from the image in order to create an automatic thresholding. I know that once I figure this out, I could go to (Analyze, Measurements, Set, Limit to Threshold)

I have also considered using the (Process, Find Edges) tool. First, selecting a box area for the program to work in, then finding edges, allows the program to pull out the nerve fibers. I can also then (Process, Binary, Make Binary). However, I do not know if making binary will assist me in quantifying the density of nerve fibers. Next, if I (Analyze, Measurements) I can then get the Integrated Density, which takes the product area*mean gray value. I am not sure what this is actually telling me. I noticed using this method, the min and max numbers where different for each photograph, therefore the IntDen value is not significant, however, if I could set the threshold values in some way, then would this calculation of Integrated Density tell me the density of nerve fibers in my selected area?

Thank you again. I look forward to getting this problem solved with your help!

Sincerely,

Lacey Beth Elwyn, MS-II
Chicago College of Osteopathic Medicine
Midwestern University - Downers Grove, IL

 
Lacey Beth Elwyn, MS-II
Chicago College of Osteopathic Medicine
Midwestern University - Downers Grove, IL