Hi! I am trying to measure fibre length in several of regions throughout hippocampus in sections stained for dopamine beta hydroxylase imaged in bright field at (20x, 40x, and 60x) and was wondering if anyone has a protocol that they could share? Preferably, it would be for MCID and ImageJ, but I am open to exploring other softwares as long as they are free. Thanks!
Use ImageJ. It has a tool that allows you to trace a line freeform. If you want length, you zoom in, and trace the fiber, keeping the cursor in the middle of the fiber. The measure function will tell you the length of your traced line.
download image j/ Fiji, go to the plugin, use simple neurite tracer.
https://imagej.net/Simple_Neurite_Tracer
I am very familiar with Image J/ Fiji. You may need to follow these steps:
1. File > open > select image (open image)
2. Image > type > 8-bit (change to B & W)
3. Process > subtract background > rolling ball radius
4. Image > adjust > threshold (B & W)> apply
5. Analyse > analyse particles
a. Area
b. Perimeter
c. Circularity
d. circumference
e. Feret's diameter
6. Exclude on edge. show outline, show clear results, display results
7. Transfer to excel
I hope that this can help.
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