Posted by
Julian Smith III on
URL: http://confocal-microscopy-list.275.s1.nabble.com/fixation-of-flatworms-for-confocal-microscopy-tp722505p722524.html
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalOur worms are much smaller (1mm x 200µm), but we're successfully using
4%PFA in 1x PBS with 10% sucrose and use triton X-100 for permeabilization.
If your worms can be anesthetized, this should work fine.
Julian
Xinyu Zhao wrote:
> Search the CONFOCAL archive at
>
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal>
> Hi, everybody,
>
> We have a user trying to do confocal microscopy on flatworms, ( parasitic blood flukes with a size of 1mm x 1cm, Schistosoma mansoni, ). The goal sounds easy: imaging the nucleus of the worms. And I thought the common procedures for immunotaining tissues would apply, i.e., PFA fixation followed by Triton 100x permeablization before the staining.
>
> However, in the handful literatures available, people seemed to always use something else, such as an alcohol-formalin-glacial acetic acid solution.
>
> Can anybody explain why ? And has anybody tried just PFA and Triton 100X on these creatures?
>
> Your input will be very much appreciated.
>
>
>
> Xinyu Zhao (Jasmine)
> Biomedical Imaging Core Lab
> Department of Pathology and Lab Medicine
> School of Medicine
> University of Pennsylvania
> Tel: 215-898-6730
>
>
--
Julian P.S. Smith III
Director, Winthrop Microscopy Facility
Dept. of Biology
Winthrop University
520 Cherry Rd.
Rock Hill, SC 29733
803-323-2111 x6427 (vox)
803-323-3448 (fax)
803-524-2347 (cell)