Mounting multiple mouse spinal cords for cryosectioning - How do you do it?

Following on the success of this thread

I want to open up a similar one about working with spinal cord.
For making thin mounted sections (14 um) for things like RNAscope, I have previously placed up to 4 spinal cords in cryomold with OCT and then mounted the mold perpendicular to a chuck to make transverse sections.

I directly mount these to a Superfrost slide.

It works OK and I can put several sections per slide. The issues I run into though are inconsistency. Not all slices also mount flat and I also get an issue with air bubbles underneath the tissue section as its thaw mounts to the slide.

I haven’t come up with a great solution to this. I remember @esypek asking about how to avoid bubbles a while back, and I didn’t have a good answer then either.

What is your practice for making thin thaw-mounted cryosections of multiple spinal cords?

@adgaudet @tberta @ShanTan @sshiers @liz @runDRG @Amaury @marvizon @CandlerPaige

I posted a video a few years back about my method. It’s worked pretty well but I want to see if there are improvements I can make to get more sections perfectly cut without bubbles and folding.

I’ve never had an issue with bubbles in my slide mounted slices, but folding is one of those things that I’ve just accepted will happen every now and then.

I flash freeze spinal cords directly into OCT, but only 1 SC per mold and have never had any major complaints with that method. The single SC per mold might be a little bit more work intensive, but I think it’s easier to position the slices and control for things like folding.

@sshiers might have more insight - she’s the IHC expert!

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