Hello everyone, I have a question about spinal cord Immunoflouroscent staining with frozen sections, Is there any way to keep the spinal cord orientation still known? (right side and left side). In my study, I need to keep the sections with known orientation from dissection until imaging, at least during imaging i need to mark one side to know which one is right and which is left. pleas if any supportive suggestions, your suggestion is appreciated.
thanks
Floating or slide mounted?
Do you care about the ventral horn?
@tberta taught me a trick. After you’ve mounted your cord in OCT (where you should know the orientation from how you mounted it), you can take a small needle (22g insulin syringe) and make a small hole in the ventral horn of a known side. Then you can see the hole on microscopy and know the side.
If you don’t want to damage your tissue at all and want both ventral and dorsal intact, I suggest using slide-mounted sections. However, you will need to be perfectly consistent in how you mount your sections on your slide so that you always know the orientation, since there will be no marking. You will just need to trust that left is left and right is right, based on how you oriented your sections coming off the cryotome.
@liz might have some more tricks
1 Like
Great, I will use the needle trick, even I use slide-mounted sections still better to mark one side. Thanks👌
This was going to be my exact response - if there’s a region you don’t care about (such as ventral horn), use a pin or needle to make a hole as Alex described.
2 Likes
You can also try a colored tissue dye on fixed tissues in the less important area like the contralateral ventral horn (if you have an ipsilateral treatment). Often you can see it grossly depending on how liberal you are, which is helpful.
@hlrossi Thanks for the tip. Can you elaborate? Like inject a little bit? Methylene blue? What kind of needle?