If you need to label living schistosome cercariae to track them intravitaly, what stain would you use?
Can you clarify, by tracking intravitally, do you mean you will be observing them during the "worm tour" after penetration? Are your desired results more than pinpointing location at different times, or do you wish to observe some morphological or biochemical transition? Are you hoping to tag then follow, or just do vital staining in situ?
These points are important because uptake and retention of the dye can be problematic, particularly if you try to label in the sporocyst (once fully embryonated, they seem rather dormant). I used a variety of fluorescent dyes on six species of schistosomes for my doctoral thesis, and the surprises were nonstop: schistosomes aren't mammals.
I wonder which fluorescent dyes are suitable for cercariae staining before penetration and which would be detectable at least 2-3 days after migration in the host (e.g., if I dissect the schistosomula out). I don't care what structures they tag, I just need to see the worm as whole. I have failed to search for any paper on this topic but the data seem to be rare. I've just found some papers about utilizing succinimidyl-esters.
Thank you for advice or directing me to some literature!
Although I have no experience with yeast, I would suggest DiI, DiO or DiD (from Lifetechnologies) depending on your Filter set up (but with this order of preference). These dyes are easy to use, rel. cheap and long time stable.
Best wishes Soenke
Thank you Sönke, I have tried DiI yet, but it did not work, likely because cercariae have a thick glycocalyx coat on its surface preventing DiI binding.
As a neuropathologist I don't have experience with cercariae, but I would llke to indicate the article "Thornhill J et al. Schistosoma mansoni cercariae experience influx of macromolecules during skin penetration. Parasitology 136:1257-1267, 2009." The authors have used intravital fluorescent staining to study cercariae.
you could try mCLING:
http://jcb.rupress.org/content/205/4/591.long
https://www.sysy.com/products/mcling/gen.php
But I do not know whether it will stay on the membrane for so long...
I'm not sure if you're still after an answer, or whether it will work for what you are after, but there's a technique I've used in the past for tracking cercariae which involves a fatty acid analog fluorescent dye. The details for it can be found in these two publications.
Article Genetics, intensity‐dependence, and host manipulation in the...
Article The use of fluorescent fatty acid analogs as labels in trema...
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