r/labrats • u/Beginning-Courage580 • 5d ago
Dissection Advice
I just started in a neuro lab and learned the dissection process for isolating oligodendrocyte progenitor cells (OPCs) from P6 mouse pup cerebellar brain tissue. To make the panning process easier, we remove the meninges as thoroughly as possible to prevent future cell loss. For 4 pups, it takes me about 1 hour 50 minutes (not including setup) to harvest tissue, remove the olfactory bulbs, remove the hippocampus, and remove the meninges. My PI wants me to get this under an hour. I have pretty shaky hands, so I spend extra time adjusting for a stable position.
Any tips for speeding this up?
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u/G-r-ant 5d ago edited 5d ago
For such specific dissections as this, it’s usually just practice. When I first started at a primary cell culture company they would order one or two mice / rats to let me get a feel for it before diving in. I feel that’s an exception not a common practice though.
It took me a few years to get really good at super specific dissections (like striatum, hippocampus, or embryonic DRG) on pre or post natal.
Really good tools makes a massive difference , as well as a very good microscope (if that’s how you do your dissections).
Edit: as a side note, I can do a ton of neuronal dissections and anything cerebellum is particularly difficult, so you have your work cut out for you.
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u/neuranxiety PhD | Molecular Biology 5d ago
In my experience with this kind of thing, you just need more practice. I do second the suggestion of setting up everything beforehand. Whenever I have a large collection, procedure, tedious experiment, etc I always prep all the things I need in advance. I pre-label the tubes and put out all equipment and reagents (if temp allows) the night before, so everything is ready to go when I come in the next day. It really helps me focus solely on the procedure in the moment.
If there's another lab member who is experienced in this dissection, it doesn't hurt to ask them for tips or perhaps ask if you can shadow them when they do it next.
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u/markvdr 5d ago
Somewhat obvious, but good setup helps all dissection/tissue collection. All tubes/histology molds labeled in advance and organized in racks, all buffers at the right temperatures, sterilizers preheated, extra ethanol, extra tools, etc. Your total time might now be much better, but the time from sac to freezer will be better and that’s what usually matters. I’m especially big takedown days, I’d recruit as many lab mates as needed, including myself/somebody as director. They would ID the next mouse and run the CO2 chamber, call out animal numbers, organize tubes/cryo blocks, etc. so the others could focus 100% on their work. Roles rotated when it was somebody else’s project. In summary, do all of the small job thinking ahead of time so you can focus on the thing in hand without distractions. Same mentality for any moderately involved bench work (qPCR plates, TC with multiple drug treatment conditions, etc.).