Enzyme method for clearing and staining fish skeletons for study
Modified technique of the enzyme clearing and staining procedure
designed specifically for Ichthyology students.
For specimens (<80mm S.L.), fixed in formalin and subsequently
stored in 70% EtOH.
Method
(Sept. 17) (Steps performed for you)
Dehydration:
1. Transfer specimens to 100% EtOH..............................1
day
2. Transfer to new 100% EtOH........................................1
day
Lab 2: (Sept. 17-18)
Cartilage Stain:
3. Transfer to alcian blue solution....................................24
hr
Day following Lab 2 (Sept. 18-19)
3a. Discard alcian blue solution
Neutralization:
4. Transfer to sat'd NaBorate solution..............................~1
wk
Lab 3: (Sept. 24-25)
Bleach:
5. Transfer to bleach solution.............................................1
hr
Digestion:
6. Transfer to trypsin-buffer solution................................1
wk
Lab 4: (Oct. 1-2)
Bone Stain:
7. Transfer to alizarin red solution....................................1
wk
Lab 5: (Oct. 8-9)
Destain:
8. Transfer to 1% KOH solution.......................................1
hr
8a. Transfer to 1% KOH solution......................................1
hr - 1wk
Lab 6: (Oct. 15-16)
Preservation:
9. Transfer to 40% glycerin solution................................1
wk
Lab 7: (Oct. 22-23)
10. Transfer to 70% glycerin solution...............................1
wk
Lab8: (Oct. 29-30)
11. Transfer to 100% glycerin solution.............................1
wk
Lab 9: (Nov. 5-6)
12. Transfer to 100% glycerin solution plus
thymol crystals.......................................................Complete
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