our method was fixed in 10% NBF and finally placed in 70% ethanol, worked well

Papers enclosed

Hi Stephanie,

I have used protocol where bones are fixed in 10% Formalin before uCT scans. It works perfectly.

As you mentioned snap freezing bones in liquid nitrogen might introduce micro cracks. I would not recommend it specially if you are looking for bone marrow measurements. However, if it is difficult to obtain new bones, there is no harm in trying few snap frozen samples (just to get an idea).

Good luck!

Hi Stephanie,

We have two methods for bone sample preservation and storage depending on the intended analysis:

For micro-CT scanning and histology analyses, we incubate the freshly dissected mouse bones in 10% formalin for 24h in the cold room. Next day, we rinse samples extensively in tap water, transfer to 70% ethanol, and store in the cold room until analysis. If we need to ship these samples, we ship at RT for overnight delivery.

For biomechanical testing, we just drop the freshly dissected bone in a tube containing 2 ml PBS and store at -80 until analysis. We do NOT snap freeze the bones before freezing. If we need to ship these samples, we take samples out and ship on dry ice for overnight delivery.

I hope this helps.