embedding for immunofluorescence

Michal Opas m.opas at utoronto.ca
Sat Oct 28 12:28:19 EST 2000


Due to all sort of difficulties I am not sure anymore if this reached the
forum. My apology if it is a re-posting.


Dear All,

While my question is only partly related to fluorescence/confocal/microscopy
I feel that this might be a right forum to pose it.


We require a procedure that will reliably provide good morphology and
immunocytochemistry (preferably fluorescence detection-based) of embryoid
bodies differentiating from embryonic stem cells. In our hands, embryoid
bodies reach up to 1 mm in diameter and, as expected, comprise several
tissue types.
Ian Gibbins has been kind enough some time ago (August 14, 2000) to describe
a polyethylene glycol (1,000 MW /  1,450 MW) embedding method together with
fixation procedures they are using for neural tissue.

In addition to polyethylene glycol, several people have been using polyvinyl
alcohols.  I would love to receive comments regarding applicability of
either medium to sectioning and immunolocalization.

Finally, products such as ImmunoBed (Polysciences) and Technovit 8100
(Heraeus Kulzer) have been also advertised as extremely suitable for the
aforementioned tasks.

Predictably, a novice such as myself, is getting utterly confused trying to
sort out what should be tried first and what not at all.

Any practical advice shall be highly appreciated.
My apology to all of you who are not interested.



     Dr. Michal Opas
     Cell Biology
     University of Toronto
     1 King's College Circle
     Medical Sciences Building, rm. 6326
     Toronto, Ontario, M5S 1A8 Canada
--------------
                phone: (416) 978-8947 (laboratory)
                            (416) 971-2140 (office)
                     fax: (416) 978-3954
                e-mail: m.opas at utoronto.ca
www homepage: http://www.utoronto.ca/anatomy/opas/start.htm


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