Slice culture: Preserving circuitry in culture
As many of you know, my experimental background is in hippocampal culture. Recently, I attended a hippocampal slice culture workshop given by members of the Hayashi lab here at MIT. I never really knew too much about the pros and cons of slice culture. After seeing the technique, I wrote up a little summary of the major differences from the point of view of someone who uses culture:
- slice culture can be done quickly. if you’ve got the mediums made, it takes 10-15 mins start to finish!
- hayashi lab uses P7 rats. Anywhere from P0 to P10 is viable for slice culture. Younger is better for certain genetics work (eg. transfection with gene gun). at P7, you get about 20 slices/hippocampus.
- P7 rat hippocampus can be dissected with only the aid of a magnifying glass! It’s macroscopic.
- coronal slicing results in a mostly intact hippocampal circuit: DG->CA3->CA1. In vivo, synapses form at P10.
- slicing is done with a tissue chopper. a vibratome is too slow (faster = more viable slice culture). 300-350um slices are used for patching and/or imaging. you can go thicker for imaging-only.
The biggest advantage over culture seems to be that you get an intact-ish hippocampal circuit. The biggest advantage over acute slice is that you don’t need to slice every day (and wait for recovery 1 hour post-slicing). Neat technique.