I am wondering if this is a technical issue that occured during processing. I have murine femurs that were decalcified in 15% EDTA over 2 weeks and then processed and I embedded them into FFPE. Before I started to section the bones I noticed what looked like air bubbles around the bone. I melted the blocks and determined that the air pockets are actually within the shaft of the bone (2-3mm in length). Does anyone have any suggestions on how they might have gotten in there and if there is a way of removing them without damaging anything?
The source of the bubbles is either some outgassing due to high temperature or air trapped in the porous structures of the bone. You can try the following order:
1. Apply ultrasound when bone remains in liquid to perform ultrasound degassing.
2. Try cryosectioning instead to solve the problem if the gas if formed due to the paraffin heat.
3. If the first and second approaches do not remove the bubbles, try to use cryosectioning and place the mold into a vacuum chamber before freezing it to perform vacuum degassing.
Thank you for the suggestions. I am little nervous to try the ultrasound to degas, could this cause the damage to the marrow structure. I will try the vaccum degassing to see if I can make that work.
- Badges
- Science topic
- Similar topics
- Names