Dear colleagues,
I would like to known what is the best method for making permanent mounts to study mophologically and morphometrically under LM bigger nematode specimens (more o less) 25 mm).
Thanks,
CGG
Generally, it is not recommended to make permanent mounts for large nematodes. Nematodes should be studied between a slide and a cover glass with little pressure (in glycerin or another medium) and you should be able to move gently the cover slip to see all sides of the nematode body.
I agree with Jean-Lou Justine, permanent mounts of nematodes are not standard. Since nematodes are usually cleared and mounted in water-soluble agents, long durability of a permanent mount is not guaranteed. The most common agents to clear nematodes are glycerin, lactic acid and lactophenol. The latter may work better with large nematodes but it is a health hazard. So I'd use it only if nothing else works.
Hi colleagues,
Juan Antonio Balbuena
Jean-Lou Justine
I am working with ascarid nematodes. Part of these nematode are deposit in ethanol 70%. Can I use these nematodes? Can I pick directly up the nematodes and put in lactophenol?. Do you have any protocol (step to step) or article descripbing this protocol? Please :)
Yes you can. If they are large, leave then at least 24h in lactophenol. Use the same agent to mount the speciments between a slide and a coverslip. You can return the specimens to 70% ethanol when you are done.
Very large nematodes need a really good clearing agent with a low refractive index. I recommend beechwood creosote following a period in 80% ethanol to remove water.
You can use either wax method or use wire of suitable thickness to avoid flattening or you can break a glass slide and select suitable size.
beechwood creosote and lactophenol are toxic, thus such protocol would be risky for technicians or your students. Anyway, if you have enough specimens it is better to make permanant slides from part of them and other part keep in glycerin in a tube; during storage the permanent slides may detiorate or dried over 10-20 years. Glycerine suspensions are more stable in a collection storage.
- Badges
- Science method