I am working on L4, L5 DRGs - but somehow after sectioning them - I do see nerve fibers in my DRG section. Can anyone suggest me how should I get a section which does not have much nerve fibers in it. All of your suggestions will be greatly appreciated.
I think this inevitable. It depends on where you are in the section. The nerves do run through the DRG, as you know.
I think though that when people really want to see a lot of the bodies, they will mount the DRGs with the long axis perpendicular to the blade, rather than the long axis parallel to the blade. Either way, you will see axon. Those pretty publication images where you see tightly packed DRG bodies… that’s definitely not every section.
I agree with @achamess - there is going to be fibers no matter what. I usually try to trim off the roots when I dissect DRGs, but there is always going to be small pieces left attached and of course, there are fibers running through the bulb portion as well.
When I section, I take a “not for keeps” section and place it onto a slide, and then check it under a bright field microscope to make sure I can see cell bodies. I toss the slide after since its a thawed out fresh frozen section at that point. Some times, the sections are just purely nerve root which means I need to keep cutting until I get to the cell bodies. I absolutely hate wasting the oh-so-expensive RNAscope reagents just to find out its a section with only fibers and no soma… doing a 2-second check under a light microscope fixed that problem though!
Thank you. I will take that into account.