production of epon mixture
|Affiliation||Department of Cell Biology, Graduate School of Medicine, Osaka University|
|Contact||inquiry about this protocol|
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- Spread alminium foil. Pour the indicated amount of Epon812, DDSA, and MNA into the measuring cylinder on the alminium foil. *be careful not to have the reagent touch the wall of the measuring cylinder for accurate measurement.
- Plug the measuring cylinder, seal the joint with parafilm, and mix the reagents well by rotating the measuring cylinder manually for 3-5min. Fix the measuring cylinder on the EM- Infiltrator with vinyl tapes and mix by rotation for 30min at speed 30 (slow speed).
- After mixing the reagents completely, measure DMP-30 by a disposable syringe and add to the mixture, and then shake the measuring cylinder very fast and vigorously for 5min to mix the reagent as completely as possible. * If you are slow at shaking at this step, DMP-30 enhance polymerization of only a part of the epon so that the epon cannot be used for EM.
- You can use the epon after it is completely mixed. For long-term storage, you can store it at -80 degrees. In case of long-term storage, you should remove the epon completely on the wall inside of the plug and seal the joint of the measuring cylinder by parafilm.
- When the measuring cylinder becomes empty, wash the inside with acetone. The first waste should be put in the epon waste storage. This epon should be discarded after polymerizing in the oven for the waste. The empty measuring cylinder can be reused after it is completely dried up.