[Zbrafish] Re: ZebraFish Skin Histology

Katy via zbrafish%40net.bio.net (by kbaden from gmail.com)
Fri Jul 25 11:48:02 EST 2008


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.



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