You can use a bath of Isopenthane, cool it down to -50/-70 C using liquid nitrogen (do not add it directly to the Isopenthane). Have some tissue molds, containing your brains, on dry ice, and when the bath has reached the right temperature, very quickly place OCT on top and dip them in the Isopenthane. If you're quick enough your tissue should not have too much of a heat shock.

Thanks for your quick reply! I'll try it.

We section snap frozen samples for IF often in our lab. We do use OCT for this, but totally different from the protocol that Fiorelli describes.

I our case that is skin biopsies. I wouldn’t know why it won’t work for other soft tissues

A drop of water is pipetted on top of the metal plate (holder?).

On top of that, put a small piece of filter paper

The rest of this procedure is performed in the cryostat cutting chamber

Let the holder cool down for a minute

A ring of OCT is now put on top of the filter paper, leave to freeze completely

Bring some OCT in the centre of the ring, Immediately stick the tissue in the centre drop of OCT (still fluid). Leave some of the frozen tissue stick out of it.

Freezer spray is sprayed over is as soon as possible and leave for a minute.

Now you can start cutting

Good luck

Thanks. I guess skin would behave very differently than other internal organs. Do you KNOW that this method don't work for other tissues, or you just wondered if?

I don't know for sure if it works with other tissues, I never did but. Maybe some other RG-user knows. It is certainly easy to perform, just give it a try if nobody fill this in.

OK, thanks!

can you put a biopsy directly into OCT right from the surgical room?

thanks but that is the problem, I don't have a fresh tissue, I have a frozen one. Thanks anyway!