I'd like to learn how to prepare horizontal mouse brain slices to patch CA3 pyramidal cells and, preferentially, CA1 pyramidal cells and DG granule cells. I've seen different protocols, and I'm not sure which to choose. Should I make the "magic cut" (i.e., separate the hemispheres and then cut the dorsal side of each hemisphere at an angle) or just glue the whole brain the ventral side down without cutting it or something else?
I assume horizontal and transverse mean the same thing. Please correct me if I'm wrong.
The 'magic cut' is definitely the choice for getting nice ventral hippocampal slices for patching in dentate gyrus and CA3. I have not tried patching in CA1, but my guess is the 'magic cut' will also work for ventral CA1. If you are interested in the dorsal hippocampus, I would not recommend 'magic cut'.
For CA1 cells, honestly, I don't see the need for a "magic cut". my experience patching CA1 PCs from either Coronal or Sagittal planes works well. Never tried Horizontal but I bet you, it might work just fine. Bottom line: Quality of brain slices is what matters most. Good luck!
Sorry for the late reply:
So, there would seem to be an important consideration in choosing in what plane to cut a slice for your experiments: What, if any, circuitry to you want to include (e.g., mossy fiber inputs to DG? Commissural/Associational to CA1?...). In any case, you might consider dissecting out the HF from cortex, which gives you a lot of flexibility in terms of planes-of cut (of course you'd have to use a "chopper" rather than a vibratome and could not freeze it to the stage).
Hi there
I have done magic cuts for patching hippocampal CA1 cells from the dorsal, intermediate, and ventral lobes. It works fine without any issues. You can check papers from John Gray's lab at UC Davis.
John A. Gray, M.D., Ph.D. - Google Scholar
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