Myopathy with IgM binding to Decorin
Snap frozen human striated muscle. (Use the
isopentane freezing method previously described.)
use snap frozen tissue.
Technique: Cut 10 - 16 micron (12 µm)
sections in cryostat from snap frozen biopsy. Attach one or more sections
to a No.1½, 22 mm square coverslip
1. Place the coverslip with section in a ceramic
staining rack (Thomas Scientific #8542-E40).
2. Immerse sections in the filtered Harris
Hematoxylin for 1 minute.
3. Remove rack to a beaker with tap water.
4. Exchange tap water until the water is clear.
5. Immerse sections in EOSIN stain for 1-2
6. Remove rack to a beaker with tap water.
7. Exchange tap water until the water is clear.
8. Dehydrate in ascending alcohol solutions
(50%,70%,80%,95% x 2, 100% x 2) in columbia staining dish(jar)s
- Thomas Scientific #8542-E30 .
9. Clear with xylene (3 - 4 x ) also in columbia staining dish(jar)s - Thomas Scientific #8542-E30.
10. Mount coverslip onto a labeled glass slide
with Permount or some other suitable organic mounting medium.
Nuclei and other basophilic structures are
blue. Cytoplasm and acidophilic structures are light to dark
1. Thompson, Samuel W. SELECTED HISTOCHEMICAL AND HISTOPATHOLOGICAL METHODS, Charles C. Thomas, Springfield, IL, 1966.
2. Sheehan, D.C. and Hrapchak, B.B.: THEORY
AND PRACTICE OF HISTOTECHNOLOGY, 2nd Edition; Battelle Memorial
Institute, Columbus, OH, 1987.