We have muscle tissue sections (approx. 8 μm width) object slides, which are freeze damaged.
Anyone aware of a recovery protocol in regards to freeze damage, which works with sections already cut and put on object glass?
If you can figure out how to solve that problem, you'll be rich and famous. The artifact usually occurs when the muscle is being frozen, i.e., it freezes too slowly, at least according to the cryostat gods I know.
Hi Gordon
Thanks for you reply - yeah I fear that it is a long shot. But may be I should pursue a solution a bit more (cf. rich and famous).
Best, Jakob
sorry to hear, Jakob, but once you have freeze artifact, then it's all over for your tissue. As said above by Doc Warren, freezing slowly causes this, so snap freezing on liquid nitrogen is best; there are various ways.. I had the unfortunate incident years ago where I snap froze many rat brains in Liquid N2, according to the standard method. Everything was fine with the tissue until i stupidly stored the brains in a -20C freezer that had a defrost cycle... this thawed the tissue and slowly refroze it rupturing all the cells, making it look like swiss cheese under the scope. so, moral to this tale, don't just freeze rapidly, also store you tissue in a -80C freezer, or at least one without a freeze/thaw defrost cycle! Good luck
Thanks for the answers.
I am aware of the protocols regarding avoidance of freeze damage (fast freezing, sucrose etc.) - so it is not a general problem, but a problem related to a specific case, where we did not have the optimal solution when harvesting the tissue material.
Best, Jakob
Your muscle sample is physically damaged, there is no recovery from that.
The only hope for you is, that there is some area in your tissue, that is still reasonably intact. Sorry!
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