About NAN. I had tried many times, but still get NAN!

[kimjinou]

I am conducting tests on the vitality of the swimming spores of kelp.
I meet a few questions.
Firts, unsupported compressed. I use Virtualdub save my videos to uncompressed avi, problem sloved.
Second, The NAN !
I tried to change the parameteres, but it didn't work. Can someboby help me?
Here is the videos parameteres:
Under 4X ； 30882064；30fps; 0.8microns per pixel(not sure ,because it was derivation by 19201080's 2.089 microns per pixel).

Here is the video:https://drive.google.com/file/d/1MNkb1uZ-UYKzcmGTt_MajRfjaAhAyoYX/view?usp=sharing

Here is a processed videos ：（it cloud used in casa but i found it may not right):https://drive.google.com/file/d/1ZdWhqyNAUA-mK8-5Hz2TfycCF7SpiN4D/view?usp=sharing

Hope you could give me some advices.
Sincere thanks！

[kimjinou]

New thing to add!
I tried again with a 10X videos.
I found fellow this parameters could get the results.
But I think it might be wrong， because these tracks looks like not belong to sperm. And many sperm didn't get tracks.

Here is introduction about what am i doing.
We want to combine the motility and viability(how many kelp's sperms can move) to test the diffrent kelp fertility to guide the proudct. So we really want to know how to use Opencasa well.
Sincere thanks!

                    [ Here is the 10x videos:  https://drive.google.com/file/d/1j7G4XfrwR-o0o-FWqpLNB5MIkuYTwBVq/view?usp=drive_link

here is another do therload videos: https://drive.google.com/file/d/17Oo59OD-hi_aiAGHJkMHfNpCpnW_Slxz/view?usp=sharing

[kwizjunior05]

I am having the same issue.

I have spent several days trying to get OpenCASA to run, but I still get 0 and NaN for all motility estimates using the supplied test data and parameters from @calquezar. I understand this is a known issue, but the troubleshooting steps supplied on the google group are not working. Below are the notes for everything I have tried. Unless otherwise noted, the issue is present identically for all versions.

Machine specs:

• Windows 11 Enterprise (version 24H2)
• 64GB DDR5 RAM
• Intel(R) Core(TM) i7-14700F 2.10 GHz

I have attempted the following versions (downloaded from here) with both v1 and v2 of OpenCASA:

• fiji-win64: Latest version of Fiji for windows 64 bit
• ij149-jre8-64 (ImageJ 1.49 bundled with Java 8 runtime for 64 bit operating system)
• ij149-nojre (with Java 21 installed and freshly updated on my system)
  • This ran into a different one-time issue where OpenCASA was unable to create the settings.config file due to a permissions error
    • OpenCASA seems to attempt to create this file in the Java runtime location
    • Closing and reopening fixed this
• ij152-win-java8
• ij153-win-java13
  • This version caused a "Log" window to pop up saying "ParticleAnalyzer: threshold not set; assumed to be 255-255"
  • Curiously, I did not see this in any other versions
• ij154-win-java8
• ij154 (cross-platform; Java 21 installed and freshly updated on my system)

I have also attempted running it on on WSL Ubuntu 24.04 LTS, with the latest version of ImageJ (installed with sudo apt install imagej). I was unsuccessful.

Troubleshooting notes:

• All versions not bundled with a runtime executable required an extra step of telling ImageJ where to find the executable, closing, and reopening (as noted on version 1.49 above)
• I always placed javaml-0.1.7.jar to the ImageJ/plugins/jars folder.
• I always placed OpenCASA_.jar in the ImageJ/plugins folder.
• I always closed and reopened the application.
• I always used the test data and parameters supplied here
• I always unzipped to my Documents folder and ran either the bundled executable or the run.bat file to ensure using the correct Java runtime.
• I have restarted my PC several times.
• I have installed and updated Java on my machine and am currently using Java 21.0.6 installed from here (I installed the JDK because I am a developer)

Short of building both ImageJ and OpenCASA from source (which I doubt would fix the issue), I am at a loss for what to try next.