Tendon and Tissue Alignment Quantification

Hey Image SC community,

I am running some analysis on histology tissue slides containing tendon and collagen. They are stained with Masson’s Trichrome where the tendon and collagen show up blue (and sometimes red if loaded). From this larger image, I am quantifying several measurements, including tenon alignment.

Previously, I have been using a plugin for this (TWOMBLI) but I am not seeing the results that I had intended. I have been introduced to the Directionality analysis on Image J which may be my new favorite but I wanted to reach out to the forum to see if anyone has used a better macro or tool for this process.

These images are less than 100MB and are of higher resolution and I would prefer to keep them that way. Here is an example of a highly aligned tendon:

Using the TWOMBLI analysis I was given an alignment around 50-60% which I thought was low since this is a very aligned tendon.

Using the Fiji directionality I got these results from the same image.

I think this is a fine representation of alignment quantification but also wanted to get some advice from the experts. Is there another tool I could use that has been tested?

Thanks in advance for any help and advice!

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The only thing I might add here is that if you get some sort of measurement from a FIJI plugin that can run through QuPath, you might be able to create tiles that report the results of the FIJI plugin’s measurement (as a measurement passed back to QuPath). The plugin would have to be ImageJ1 compatible though, I think.