In summary, the brain goes into a mixture of solution A and B (1:1). Brains stay there for 2 weeks in the dark. Then they are transferred to Solution C for 1 week. And as suggested by the company I have been using solution C for sectioning the brain in a vibrotome.
Sectioning the brain in a vibrotome requires a lot of volume from this solution C.
Originally I started using sucrose (30%) to section the brain, but once I looked at them under the microscope there was a lot of black fungi or mold looking particles all over, and they grew more and more over time.
I called the company they told me to follow the manual and use solution C for sectioning the brain and collecting all sections.
It is a lot of volume and we need to find another solution where we can section the brains.
Anyone has tried to use this staining kit but sectioned the brain not in solution C?
Best,
Itza
No, still use the traditional and the Cajal modification but:
It sounds like you are filling the entire bath of the vibrotome with the solution C. When we sectioned live tissue, we developed an alternated strategy. Instead of using the well of the vibrotome as a tissue bath, we got a small plastic petri dish. On the bottom of the petri dish we attached a chuck to go in to the holder. Then we attached the tissue to the petri dish (with super glue). The incubation media was then placed in the petri dish and as we sectioned, we collected the sections in the petri dish. This might help.
But there are are other alternatives to sectioning and storing the tissue. You might use a sliding microtome. -- If your tissue is hard enough. In this case, with your collection brush, you can keep your block moist with your solution C and collect the sections into the solution C. There are other solutions in which you can store your brains. Check out the cryoprotection solutions. These should work if your sucrose did. Final, it might be said that, Golgi, himself ,when he began sectioning ,used nothing more than a razor and a hand held support for the tissue. It your have a classic reaction, if should survive ETOH, xylene and other solvents. Good luck
Thanks Frank,
I do use a petri dish, and I glue the brain to it. It takes around 60mL of solution C.
I am afraid to go back to sucrose, and not sure if I should try distilled water.
Thanks for your feedback,
Best,
Itza
- Badges
- Science method