Alizarin Red Sand Toluidine Blue O
EMS Catalog #: 26205-Series
- Wash the specimen in 250 ml of Ammonium Alcohol (#26205-03). For 24 hours.
- Stain in Toluidine Blue Solution, 0.25% (#26205-02) for 1 week.
- Harden and de-stain for 72 hours in 4 changes of 95% alcohol.
- Macerate in several changes of Potassium Hydroxide, 2% (#26205-05) for 5 – 7 days, depending on the size of animal. This step is facilitated exposure to sunlight or ultraviolet light.
- Transfer to Working* Alizarin Red S solution for approximately 24 hours.
- Dehydrate through 3 changes of cellosolve, 6 hours each. Reduce the time for small embryos. OPTIONAL: One may substitute the alcohol series, 50%, 80% and 90% followed by 3 changes of benzene.
- Clear by transferring to solution for 25%, 50%, and 75% methyl salicylate in cellosolve for 24 hours each. Then store in methyl salicylate.
Modification by using glycerin for clearing: Omit step 6 and transfer directly from Working Alizarin Red S solution into a series of 50%, 70% and 80% glycerin solution for 24 hours each. Store in glycerin.
|Osseous Tissue||Deep Red|
Note: The intensity of the stains serves to indicate the relative amounts of ossification and chrondogeneis, which may be taken place. The bone cartilage may be stained separated by omitting step 2 for bone or step 5 for cartilage.
Dawson, A.B., Stain Tech., 1:123 – 4, 1926.
Williams, T.W., Stain Techn., 16:23-5, 1941.
Clark, G., (ed)., Staining Procedures, 3rd ed., Williams & Wilkins, Baltimore, p. 139, c. 1973.