Issue with OpenCASA Returning Only NA Values

[Léa Dasque]

Hello,

I’m Léa, working with OpenCASA for my PhD research on treefrog sperm motility at Lyon University, France. I attempted to use OpenCASA to extract motility parameters from my treefrog sperm videos, but I only get NA values. I tried downloading an older version of the software, but the issue persists.

I’d like to know whether the problem is due to the poor quality of my videos, making them unsuitable for CASA analysis, or if I’m not setting the correct parameters. The microscope was set to x400 magnification, and the camera recorded at 160 fps.

To troubleshoot, I uploaded both the raw video and an enhanced version where I adjusted the contrast to improve visibility. However, the results remain the same across all three attempts.  Here are the 1s videos : https://drive.google.com/drive/folders/1NF60BINGYGApAF8uO4Rw4b7QaSRMlQLn?usp=drive_link

I would really appreciate any guidance on this!

Best regards,
Léa

[f.martin...@gmail.com]

Hi Léa,
It seems that you need to open access to the video so we can download it.
Have you tried downloading older versions? 2020's should work. Not
sure, but some of the 1.54's. Just install the plugin on these and try
again. The NAs are typical for this faillure.
Best wishes,
Felipe

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[Felipe Martínez-Pastor]

Hi,
I could open the videos and process them with OpenCASA. I wonder if
you have a Ph BM objective (negative phase contrast). In my
experience, these kind of objectives are the ones providing good
conditions for ImageJ. I'm not sure if this is only Nikon
nomenclature.
The NA problem is not because of the videos. Unfortunately, we cannot
solve this problem at the moment, just try these older ImageJ
versions. Install the plugin, run it, and try in a current one. This
is the only way to make it run.

Best wishes,
Felipe

El mar, 11 feb 2025 a las 18:07, Léa Dasque (<lea....@gmail.com>) escribió:
>

[Léa Dasque]

Hello,

Thank you for your answers.

Yes indeed the objective is not negative phase contrast so I inverted the colors of the image on ImageJ.

I tried using an older version but it still doesn't work, this is why I wanted to know if the problem was coming from my video....

Best wishes,

Léa