Microtome must have anti-roll beside the blade and if not work maybe not adjusted. Also you could recover rolled sections if you put them in cryoprotectant solution (glycerol+ethylen glycol) then unroll the section and mount it with a narrow brush.

Hello, I'm glad you're experienced with brains. First of all, I tell you that it is a difficult tissue to handle in a cryotome. First given by the configuration of the fabric itself and second by the difficulty of adequate fixation of the fabric. Second, the brain classically removes tissue from the cutting blades and aligners quickly...very quickly. It really takes the edge off in a creepy way.

Here in Mexico we use RUITER a lot, which is a homemade but very useful formula so that the fabric does not tear and if it is folded it can be folded more easily. If you are interested I can gladly pass it on to you. Greetings.

First, fixation is the most critical step affecting slicing in the cryostat, vibrating microtome, and microtome—whole brain immersion in fixative results in poor preservation. Autolysis occurs in deeper brain structures, and slicing will be almost impossible. Even perfusion fixation without proper care may impact slicing. Remember that slicing a rat/mouse brain is usually a challenge because we have many structures with different densities lining each other, one of the reasons that makes it difficult to slice.

If the fixation is okay, you must soak the brains in cryoprotectant solution (2.3M sucrose). Change the solution every 6 hours for one day or two. Then, you can freeze the brain. I recommend you embed the brain in Tissue Freezing Medium with a plastic mold. By doing so, you will get a solid bloc. It will facilitate slicing. If wrinkling persists, you can also place the slice in a buffer-filled jar and then recover the slice in a glass. The freezing mounting media will dissolve in the buffer, and the section can be easily placed onto a glass slide. Good luck

Rita Sinigaglia Thank you for your response. I take the brain tissues out of PBS sucroze at 4 °C and then I placed the tissues on the freezing stage. Would you recommend keeping the tissues at -20°C degrees before taking the sections? If so, in what type of solution (long term solution buffer, OCT) and for how long. Additionally, is it necessary to keep the tissues at 4°C after they are taken out from -20°C before sectioning?"

Alberto Daniel Saucedo-Campos Thank you for your response. I need to mention this, I dont use cryotom and rotatory microtom. I use sledge microtome. I take the brain tissues out of PBS sucroze at 4°C and then I placed the tissues on the freezing stage. Would you recommend keeping the tissues at -20 °C before taking the sections? If so, in what type of solution (long term solution buffer, OCT) and for how long. Additionally, is it necessary to keep the tissues at 4°C after they are taken out from -20°C before sectioning?