Stain: Alkaline Phosphatase
More Staining Procedures For Plastic Embedded Tissue Patsy Ruegg, Metabolic
Bone Disease Laboratory of the University of Colorado Health Sciences
Center Denver, Colorado INDICATION: Osteoblasts
Alkaline Phosphatase can be done on GMA sections but
it is tricky. The
enzyme seems to be sensitive to heat so care must be taken not
to over heat the blocks during polymerization and not to heat
the sections onto the slides. I just let them air dry on silane
coated slides over night. REAGENTS:
Naphthol AS-MX phosphate alkaline phosphate solution, 0.25%
(Sigma Cat. No. 85-5)
Fast Blue RR Salt (Sigma Cat. No. FBS 25) PROCEDURE:
- Measure 50ml of dH2O into a beaker. Add
2ml of the alkaline phosphate
solution and one capsule of the Fast Blue RR Salt.
- Mix the solution well on a magnetic stirrer.
- Pour the solution on to the slides in
a coplin jar, without filtering.
- Place the coplin jar in the refrigerator (4C), with the lid
on it, and allow to stain over night.
- The next day, remove the jar from the fridge and pour the
solution into the chemical waste can, and rinse the slides
very well with dH2O.
- Air dry slides and coverslip with permount.
RESULTS:
Osteoblasts and certain other marrow cells stain purple/blue,
BONE is unstained. |
