[Zbrafish] Re: ZebraFish Skin Histology

alzaidan.abdullah from gmail.com via zbrafish%40net.bio.net (by alzaidan.abdullah from gmail.com)
Sun Sep 21 22:31:53 EST 2008


On Jul 26, 1:48 am, Katy <kba... from gmail.com> wrote:
> On Jul 21, 6:50 pm, alzaidan.abdul... from gmail.com wrote:
>
> > Hi,
>
> >  I’m currently studying the effect of contaminants on the skin of
> > Zebrafish through Histology. The problem I’m facing while processing
> > the tissue is after doing so the skin always detaches from the muscle.
> > The fixative used is 4% NBF for 24 hrs. And I’m only removing the skin
> > attached with muscle, also tried to cut a TS section (Dorsal to
> > Ventral through the vertebra) but similar effect. The thickness used
> > on microtome is 5um.
>
> > Any help would be appreciated
>
> > Abdullah Al-Zaidan
>
> Hello,
> Are you looking at H&E stained sections?  At the ZIRC we use
> Dietrich's fixative for all H&E specimens.  I've pasted our fixation
> protocol with a recipe for Dietrich's fixative  below.  Hope this
> helps.
> Katy Murray, DVM, Ph.D.
> Zebrafish International Resource Center
>
>     *    Fixation Protocol for Histopathology Specimens
>     * Prepare Dietrich's Fixative (100 ml) as follows. Store fixative
> at room temperature.
>            30 ml Ethanol (95%)
>            10 ml Formalin (Formaldehyde 37% solution, histological
> grade, contains 10-15% methanol, Sigma # F1635)
>            2 ml Glacial Acetic Acid
>            58 ml Distilled Water
>     * Euthanatize fish with an overdose of MS-222 (tricaine methane
> sulfonate).
>     * Carefully slit open the body cavity along the belly, trying not
> to disrupt internal organs.
>     * Use 10-15 ml of fixative per 1-2 fish in a sealable container.
> The volume of fixative should be at least 10 times the volume of the
> specimen.
>     * Store and ship fixed fish in Dietrich's fixative. Fish can be
> stored for several weeks in Dietrich's fixative.

Dear Katy:

 Thank you for your help i appreciate it.



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