Mounting

jose_jpr

After clarification, I included the sample for two days in glycerol, but doesn't sink. Also, it shows a reduction in sample size.

is it normal?

jose_jpr

If I not going to visualize it, what is the protocol to keep the sample in the best conditions?

Clara

Hi, did you wash the clarification buffer off the sample with PBST before incubation with glicerol? If you're not going to imaging you should keep your sample in PBST, room temperature or 4°C.

jose_jpr

Yes, before incubation with glycerol I washed the sample with PBST. After 4 days, the sample is sinking halfway. More time in glycerol is not dangerous for the sample? Thanks

Clara

I haven't left my samples in glycerol, so I can't tell you if it would be any problems, but I do know that you can wash your sample in PBS and leave it there after imaging, you can even continue clarifying if your sample wasn't good for imaging.

jose_jpr

Thanks for the information. After 6 days, the brain has sunk in glycerol. Now I will proceed to imaging. I hope that 6 days in glycerol, it is not dangerous for the sample and the imaging it's ok