Frog sperm tracking

[Pedro Aguiar]

Hello all,

Has anyone worked with frog sperm tracking analyses? Frog sperm typically have very elongated heads, and I suspect this might affect tracking accuracy in CASA software. I’d appreciate any advice on optimizing settings or choosing the best methods to ensure reliable tracking for amphibian sperm.  

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

Hi Pedro,
We have worked with chicken sperm (also, elongated) and no problem.
Yes, frog sperm could be tricky, but apparently it is not a big deal
if you have good contrast and the tails are not affecting the
detection. If you have problems with that, you might send an example
video and we might help with that.
As a trick, if you open the video in ImageJ and save it as AVI with
PNG compression, it becomes considerably smaller with no loss of
quality... even JPEG is acceptable and it can be <2 MB (I added this
to the group welcome).
Best regards,
Felipe

El mié, 5 mar 2025 a las 16:41, Pedro Aguiar (<pagui...@gmail.com>) escribió:
>
> Hello all,
>
> Has anyone worked with frog sperm tracking analyses? Frog sperm typically have very elongated heads, and I suspect this might affect tracking accuracy in CASA software. I’d appreciate any advice on optimizing settings or choosing the best methods to ensure reliable tracking for amphibian sperm.
>

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[Pedro Aguiar]

Thanks a lot for the suggestions, Felipe!
Currently, I do not have access to samples, but I plan to start collecting them at the end of this year.
Would it be possible for you to send me one or two videos of chicken sperm so I can visualize them on CASA? It would really help with my trials.

Best,

Pedro

[Felipe Martínez Pastor]

Hi Pedro,
You can find the video and the image with the configuration in this link:
https://we.tl/t-wmTVLzSkSd
The video is only 20 MB because I used JPEG compression (it might take
several seconds to load while decompressing).
Not very good motiltiy (thawed semen), but OpenCASA can detect the tracks.
Best wishes,
Felipe



El 5/3/25 a las 17:55, Pedro Aguiar escribió:

[Alex Price]

Hi Pedro,

I am also working with frog sperm, but I have been having trouble using OpenCASA to track motility. I am attempting to analyze Xenopus laevis sperm, and as you mention, they have very elongated heads. It seems that their head morphology and swimming pattern leads the program to detecting very jagged paths even as I adjust maximum displacement and window size.

Since your last post, have you found any parameters that track frog sperm well?

Thanks,

Alex

[Pedro Aguiar]

Hi Alex,

Thanks for your message.

Unfortunately, I haven’t yet identified reliable tracking parameters for Scinax sperm, mainly because in my experiment the sperm are non-motile most of the time. Motility is rarely observed (roughly ~20% of attempts), and so far I haven’t been able to identify a consistent pattern explaining when or why they become motile.

That said, I’d be very interested to learn more about your experimental conditions, since you’re also working with frogs. In particular, could you share some details about how you prepare your sperm samples to induce or maintain motility?

For example:

• What is the composition and salinity/osmolarity of the solution you use for sperm activation or observation?
• At what temperature do you perform the recordings?
• Do you use any specific activation step or trigger to initiate motility?
• What type of slide or chamber do you use to observe the sperm?

It’s possible that differences in these conditions are a major factor in the motility issues we’re encountering, so any insight you’re willing to share would be extremely helpful. Sorry I couldn’t be more helpful on the tracking parameters at this stage.

Best,

Pedro Aguiar

To view this discussion, visit https://groups.google.com/d/msgid/opencasa_mailinglist/6af3cdaa-bfea-4ea7-bdc5-974a240bb762n%40googlegroups.com.