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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Xinyu, we were successful in fixing different Schistosoma mansoni stages
with 3.5% formaldehyde in PBS (see Bahia et al, Parasitology , 133: 321-329
2006).
Good luck !
Renato
Renato A. Mortara
Parasitology Division
UNIFESP - Escola Paulista de Medicina
Rua Botucatu, 862, 6th floor
São Paulo, SP
04023-062
Brazil
Phone: 55 11 5579-8306
Fax: 55 11 5571-1095
email: [log in to unmask]
home page: www.ecb.epm.br/~ramortara
-----Mensagem original-----
De: Confocal Microscopy List [mailto:[log in to unmask]] Em nome
de Julian Smith III
Enviada em: quarta-feira, 13 de agosto de 2008 15:16
Para: [log in to unmask]
Assunto: Re: fixation of flatworms for confocal microscopy
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Our 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)
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