I have been trying to patch spinal dorsal horn neurons in acute spinal cord slices from adult rats. The major issue seems to be cell viability as the cells appear swollen like a balloon. I took care of osmolarity and pH; nevertheless no improvement in morphology of cells. I have tried almost all types of cutting buffers and recording buffers available in literature still did not get healthy cells to patch. I have been patching hippocampal slices also so I am wondering whether I should try to patch deep layer cells with blind method. Is there anybody who has done it or experienced that how healthy cells look like in spinal cord slices? I would like more things to discuss once I receive your suggestions.
Hi, in brain and spinal cord slices cells which are close to the slice surface(~50mkm) are usually died or not healthy, due to the cutting procedure (Nedden2011, J Neurosci). Try to optimise the visualisation methods (DIC, IR-DIC, Dodt contrast ) to have a good visibility of the deeper cells and patch them.
Hello Bhanu, we have a lot of experience recording from spinal cord slices in mice and we have found that temperature and the time you take to slice the spinal cord and prepare it for your recordings is key. Take a look at this paper from a colleague of mine who also has recorded from spinal cord slices and is using a glycerol-based artificial CSF (GACSF) that we also use. Also take a look at the temperatures used and the time taken for slice recuperation after slicing. Hope this helps... :)
- Badges
- Science topic
- Similar topics
- Methodology
- Laboratory
- Cellular Biology Techniques