Enzyme+method+of+clearing+and+staining+skeletons

=**Workshop on Enzyme Method of Clearing and Staining Skeletons**= Friday, January 30, 2015 //marie Thérèse Rush and Leslie Rye//

//Please download the following://
 * [[image:butGold1.gif link="file:Enzyme method for clearing and staining fish skeletons for study.pdf"]] Schedule for preparing your specimen


 * [[image:butGold1.gif link="file:Enzyme_Method_Clearing_Staining2015.pdf"]] Enzyme Method Information - excerpt from the Ichthyology Primer Manual



February 23, 2015 - note to students Re: Digestion - Bone Stain Steps:

Following one week of digestion process: Observation:
 * Ø If your specimen has been cleared to the point that you see the skeleton, then move to the Bone Stain Step.
 * Ø Else repeat last week’s step.
 * o Your specimen should be temporarily placed in the lid.
 * o Pour the buffer-enzyme mixture into the Buffer Waste container.
 * o Rinse out your cup.
 * o Pour new buffer solution into cup.
 * o Replace your specimen in the cup.
 * o Have Leslie (or I) add the enzyme.
 * o Wait another week for the enzyme to do its work.

For the Bone Stain step: Observation:
 * Ø If your specimen has been cleared to the point that you see the skeleton.
 * o Place your specimen temporarily in the lid.
 * o Pour the buffer-enzyme mixture into the Buffer Waste container.
 * o Rinse out your cup.
 * o Replace your specimen in the cup.
 * o Pour enough 1% KOH to cover your specimen.
 * o Add 5 – 10 drops of alizarin red stain.
 * o Replace lid and wait a week for stain to penetrate the bone elements.