Trypsin for Diaphonization
Diaphonization clears animal specimens with trypsin, then stains bone and cartilage. Lab suppliers sell the enzyme.
Updated . Edited by: Ilyass Ben Hakim.
Where to buy
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Lab and biotech suppliers sell trypsin 1:250 from porcine pancreas. Many sell to registered businesses and schools.
Hobby sellers on marketplaces split bulk packs. Ask for the grade and a certificate of analysis.
Professional insight
Museum preparators choose a crude, low cost trypsin grade for clearing. Cell culture purity adds cost without helping.
- Trypsin 1:250 is the grade most clearing protocols use.
- A sodium borate buffer keeps the enzyme working.
- Old or warm stored powder loses activity.
- Bulk 500 g packs suit large fish batches.
Check the label
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Diaphonization steps at a glance
- 1Fix the specimen in formalin, then rinse.
- 2Stain cartilage with alcian blue.
- 3Clear soft tissue in trypsin with sodium borate.
- 4Stain bone with alizarin red.
- 5Move through glycerin for storage and display.
Tools you will need
- Trypsin powder and sodium borate.
- Alcian blue and alizarin red stains.
- Glycerin for final storage.
- Glass jars with tight lids.
- Nitrile gloves, goggles and a dust mask.
Safe use
- Work with formalin only in good ventilation.
- Label every jar with contents and date.
- Keep chemicals away from children and food areas.
- Dispose of formalin by local hazardous waste rules.
Storage
- Keep trypsin powder sealed and dry.
- Store cold, as the label states.
- Mix working solution fresh for each batch.
Common questions
- Can I use meat tenderizer instead?
- Some hobbyists try papain products. Results vary, and protocols are built around trypsin.
- Is cell culture trypsin solution suitable?
- It works but costs more. Powder is cheaper for clearing.
- How long does clearing take?
- It depends on specimen size, from days to weeks.