by CombiChrist Sat Jun 12 2010, 18:42
No video, but the procedure :
Procedure:
1. Fix fresh material in 10% formalin
2-3 days
or longer. Specimens already in formalin or alcohol,
ignore
step 1 and go to step 2.
2. Wash in several changes of distilled
water
for 1-2 days.
3. Place directly into a mixture of 10
mg Alcian
Blue 8GN (supplied by Matheson, Coleman and Bell), 80
ml 95%
ethyl alcohol and 20 ml glacial acetic acid 10-12
hours. (This
solution must be fresh)
4. Transfer to two changes of 95% ethyl
alcohol,
1-2 hours per change.
5. Transfer through an alcohol series of
75%,
40%, 15% ethyl alcohol 2-3 hours in each, or until
specimen
sinks.
6. Transfer to distilled water 2-3
hours, or until
specimen sinks
7. Place in an enzyme solution of 30 ml
saturated
aqueous sodium borate, 70 ml distilled water, and 1 g
trypsin
enzyme. Change solution every 2-3 days, or sooner if
solution
turns a bluish color. Continue until bones and
cartilage are
clearly visible, and the flesh retains no blue color.
NOTE:
the skin of some gobies remains blue at this stage.
8. If bone is present: Transfer
to 0.5%
aqueous KOH solution, to which enough Alizarin Red S
stain has
been added to turn solution a deep purple-red. Leave
24 hours
or until bones are distinctly red. If bone is not
present
go to step 9
9. Transfer through a 0.5% KOH -
glycerin series
of 3/4 KOH - 1/4 glycerin, 1/2 KOH - 1/2 glycerin, 1/4
KOH -
3/4 glycerin, and finally to pure glycerin.
10. Store specimen in pure glycerin to
which a
few crystals of thymol have been added to inhibit
molds and
bacteria.