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

**URL:** <https://forum.painresearcher.net/t/mounting-multiple-mouse-spinal-cords-for-cryosectioning-how-do-you-do-it/758>\
**Category:** Experimental Techniques\
**Created:** [September 13, 2020, 2:40pm UTC](https://forum.painresearcher.net/t/mounting-multiple-mouse-spinal-cords-for-cryosectioning-how-do-you-do-it/758 "2020-09-13T14:40:42Z")\
**Posts on this page:** 2\
**Page:** 1

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**Author:** ![achamess](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/achamess/32/534_2.png) [@achamess](https://forum.painresearcher.net/u/achamess)\
**Post date:** [September 13, 2020, 2:40pm UTC](https://forum.painresearcher.net/t/mounting-multiple-mouse-spinal-cords-for-cryosectioning-how-do-you-do-it/758/1 "2020-09-13T14:40:42Z")

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Following on the success of this thread

> [@Mounting many DRG for cryosectioning - share your tips](https://forum.painresearcher.net/t/mounting-many-drg-for-cryosectioning-share-your-tips/742/11):
>
> Thanks @adgaudet. That’s also a great trick about the orientation so you know which is which. Will spare the mental spatial gymnastics I when making my slices to try to get them all the same orientation on the slide slight_smile I agree 100% with putting all the DRG that you will compare on the same section.

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.

[https://forum.painresearcher.net/t/cryosectioning-mouse-spinal-cord-and-drg-for-ihc-ish-active-users-only/198](https://forum.painresearcher.net/t/cryosectioning-mouse-spinal-cord-and-drg-for-ihc-ish-active-users-only/198)

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**Author:** ![CandlerPaige](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/candlerpaige/32/250_2.png) [@CandlerPaige](https://forum.painresearcher.net/u/CandlerPaige)\
**Post date:** [September 13, 2020, 6:12pm UTC](https://forum.painresearcher.net/t/mounting-multiple-mouse-spinal-cords-for-cryosectioning-how-do-you-do-it/758/2 "2020-09-13T18:12:47Z")

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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!