Some general "newbie" questions in regards to imaging thick samples

Hi all,

As a scientist who has only used 10-40um thick rat brain samples in the past, the idea of going to 500um-1mm thick sections is a little daunting. A couple of helpful tips would be useful.

For example, the Clarity protocol describes a tissue mounting technique for any of you use it or do you have other ways to mount the tissue for microscopy. If you have pictures to help demonstrate the some of the mounting procedures, that would be great to see as well.

The second, relatively novel concept that I am having an issue with prior to beginning working out the methods is the section of the protocol where they describe "after imaging the first round" implies that you can run your sample through multiple IHC procedures to label the sample multiple times (washing/clearing the previous abs away and using additional abs to label something else). How resilient is the sample to multiple labeling (if that is indeed what is being described?) How many times could this be performed? Again, my tissue of interest is brain...any advice or guidance?

Any resources that you all have found useful would be great as well!

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