Fixation of marine larvae

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From:Rene Hessling <> (by way of histonet)
To:histonet <>
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Dear members of Histonet,

I am encountering difficulties fixing trochophore larvae of echiurids
(marine worm-like animals) for immunohistochemical analysis and am
looking for alternative protocols for fixation.

The outer layers of tissue in late larval stages seem to burst and
dissolve after approximately 10 minutes in the fixative. Currently I am
using 4% paraformaldehyde solution in 10 mM PBS (pH 7.4), which has
proven to be a good fixative for many other marine larvae.
The pH of the fixative seems to be correct. I have varied the
formaldehyde concentration. Furthermore I have added sucrose in various
concentrations to increase osmolarity, yet without any positive results.
Also I have varied the time of fixation between 30 minutes to 48 hours
without any positive change.

I would appreciate any suggestions or protocols to help in this matter.
Thank you very much in advance.


Dipl. Biol. Rene Hessling
Universitaet Osnabrueck
Spezielle Zoologie
Barbarastr. 11
49069 Osnabrueck


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