hello;
I have a bad problem in cutting my samples. My scaffold is so dense; so baldly cut by microtom and cell layers that cultured on the top of scaffold separated from it.I could not have a good staining and finally photo for my results.
could anybody help me?
I also have no access to cryo-microtom.
thanks a lot.
Tnx Mr. Qasim, but I have no access to it in our instittute. Also I fixed all my samples.
Dear Toktam Ghasemi,
do you store your paraffin embedded scaffolds at 4°C prior cutting them? I found this very helpful, to cut it as cold as possible. Furthermore you can try laying a ice block on top of it for some minutes and cut it then. Otherwise I would try to cut different µm, sometimes this helps. The problem is really the cutting and not the paraffin embedding I suppose?
Hope this helps!
Kind regards
Dear Toktam,
I agree with the above advice, first make sure to start with a new blade, clean the stage very well and check it from time to time during sectioning. Its very important to adjust the distance and the angle of the glass cover and the stage. Start with 5-10 um thickness on a blank paraffin block until you are satisfied with the scions. Also, its helpful to put the paraffin blocks on ice from time to time particularly when sections starts to become messy. Finally make sure to clear all the bubbles at the bottom of your water bath (manually) as they might affect your sections.
Good luck
Dear Toktam,
I also agree with both advice, I put my paraffin Embedded scaffolds at -20°C 1 hour before starting to cut. I use specific blades for tough tissue.
good luck
Hi, store your paraffin embedded scaffolds at -25 ºC prior cutting them, then make the cutting process reducing environmental temperature as much as possible (10 ° C)
- Badges
- Science topic