January 8, 2018 at 5:44 am #556Emily K. BensonKeymaster
Any Biowave users out there?
Looking to give Biowave a try for staining.
It might reduce time a bit, but I think the preparation of solutions, washes, etc, changing of solutions takes up most of the time, which isn’t going to be changed using the Biowave, but management wants me to show this,… and I am happy to be wrong and for Biowave to really speed things up!
One of the major plusses I see, is that it may be much safer training people to use the Biowave, since they won’t have to take the samples (in hazardous chemicals) from hood to oven, or hood to fridge, or risk as much cross-contamination of surfaces and materials, and trust them to follow proper protocols, and be careful. (So even if the microwave settings aren’t so beneficial…the heat and cooling options can make it a safer contained unit within the hood for hazardous chemicals, and the samples won’t need to leave the hood until they are put into the oven for resin curing.)
We’re going to take a coronal slice of mouse brain, and take the corpus callosum from each hemisphere – one side will be stained the good ol’ fashioned way, the other hemisphere will be put through Biowave protocol, and then we will do a series of our standard CNS and myelin assays to see if we get the same numbers from both sides.
I came across this paper in the past, and looked it up again: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3701260/ …and it doesn’t look like the time is much reduced when using Biowave.
Not sure if anyone has any favorite Biowave protocols – I recall talking to Rick Webb from University of Queensland, who is super enthusiastic about Biowave for serial blockface staining, and was willing to share his Biowave specific protocol with me to try.
I will update in a few weeks with how it goes, but please share any experiences with BiowaveJanuary 8, 2018 at 10:51 pm #573Claudia LopezParticipant
We have used the Biowave for SBF-SEM but found that the staining is not even along the tissue. We use Biowave for all of our TEM samples with great success and for some samples run on FIB-SEM but we are still optimizing the protocol for SBF. We definitely need to modify the times used on the Biowave for each step and check if this will result in an even staining. We have tried different tissue sizes and even in the smallest one we can see like a banding pattern of staining. Once we optimize the protocol we can share it, so far it has not worked for us.
ClaudiaJanuary 8, 2018 at 10:54 pm #574Claudia LopezParticipant
Just FYI, we have tried most of our runs in pancreas, not brain.
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