LR White (LRW) Protocol
1. MixingAdd 9.9g of benzoyl peroxide to the LRW and stir for 30 minutes. Store the activated resin 24h at 4°C before use. Always store LRW at 4°C.
2. Important LRW details
– Do not store in styrene-based containers such tissue culture flasks. LRW will etch the plastic.
– Oxygen is the enemy and will prevent polymerization. Samples must be sealed airtight to use.
– Samples can also be UV polymerized in the coldroom. Use the UV handlamp on longwave.
3. Storage/Shelf LifeLRW shelf life is reported to be one year at 4°C. It can also be frozen.
4. LRW InfiltrationUse ethanol only. Do not use methanol or acetone.Allow the LRW to reach room temp (RT).
Dehydrate and infiltrate on the following schedule
- 15 min 70% ethanol
- 15 min 80% ethanol
- 15 min 90% ethanol
- 15 min 100% ethanol
- 4h 1:1 100% ethanol:LRW on rotator
- 1h 100% LRW on rotator
- 1h 100% LRW on rotator
5. LRW Embedding: Use the LRW oven (65°C). Seal your samples. Oxygen inhibits polymerization.
Beem® capsules: Remove the caps from the capsules. Press a piece of Parafilm® M into the top to seal, overlapping the edge. Recap. Polymerize overnight-48 h.