1/30/97
with Alizarin Red? We did find something for sectioned material
(Dahl's Method for Calcium) but wonder if there is a specific
protocol for whole tissues, perhaps with a tissue clearing step.
Thanks, Linda Fox
lfox1@wpo.it.luc.edu
Loyola University Medical Center
Maywood Illinois
_Staining Procedures_, 3rd ed. Pg.137ff. Biological Stain
Commission, Williams &Wilkins Co. Baltimore. 4th ed. has it also, but don't
know the page. This work is likely on a higher edition by now. Email me if
you need the detailed recipe. Hi to John!
Phil
Philip Oshel
Station A
PO Box 5037
Champaign, IL 61825-5037
(217)244-3145 days
(217)355-3145 evenings
oshel@ux1.cso.uiuc.edu
The Alizarin technique for calcium is called "Dawsons" and is quite old. We
don't have the original reference, but for bone the proceedure is: fix in
10% NFS, wash then transfer to 0.5% aq KOH to which enough alizarin red S
has been added to turn solution to deep purple, leave for 24 hours or till
bones are distinctly red, transfer thro KOH-glycerine series,3:1, 1:1, 1:3,
to pure glycerine. Store in pure glycerine plus few crystals of thymol.
Refs for meth blue plus alizarin red staining of bone may help: Bechtol
1948 Stain Technol 23, p3-9; Burdi and Flecker 1968 Stain Technol 43,
p47-48; Lundvall 1927 Anat Anz 62, p353-373. If you still require more
info, contact me via e-mail and we will try to help.
Miss A.J.Wilson
Electron Microscope Unit
St George's Hospital Medical School
Cranmer Terrace
Tooting London SW17 ORE
Tel: 0181 725 5220
1. Fix in 95% ALC for 48-72Hrs. Prolonged fix in alcohol renders tissue
less liable to maceration.
2. Remove fats in acetone for 2-4 days then return specimens to alcohol for
12-24 hrs.
3. Place in 1% KOH until bones or digits appear thro' muscle
Transfer to 0.1% Alizarin Red S in 1% KOH. (Other stains eg alcian blue, or
victoria blue can be used to counter-stain cartilage or bone at this
stage.)
4. Leave till stained or desired bone colour, change to fresh stain if
necessary.
5. CLEARING
Place in solution of 1g KOH, 20mls glycerine, 79mls dist water, leave till
sample clears.
6. When clear, pass thro' increasing conc of glycerine, 50, 70, 90, 100%
Miss A.J.Wilson
Electron Microscope Unit
St George's Hospital Medical School
Cranmer Terrace
Tooting London SW17 ORE
Tel: 0181 725 5220