SV: fixation of flatworms for confocal microscopy

Posted by Pierre Nourdine Bouchelouche on
URL: http://confocal-microscopy-list.275.s1.nabble.com/fixation-of-flatworms-for-confocal-microscopy-tp722505p722706.html

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Hi everybody,

we have just purchased an SP5 confocal trying to do calcium measurements in smooth muscle cells.
Do anybody have any experience with calcium measuremenst with SP5.

Thank you.
Best regards
Pierre
Smooth Muscle Research Center
Copenhagen
Denmark

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Fra: Confocal Microscopy List [[hidden email]] På vegne af Xinyu Zhao [[hidden email]]
Sendt: 13. august 2008 20:00
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Emne: fixation of flatworms for confocal microscopy

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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:  [phone]