Neuron counts were performed as previously described (9 (link)). In brief, torsos of E16, P0, P30, or P120 mice were fixed in 4%PFA/PBS for 4 h to overnight and cryoprotected in 30% sucrose/PBS for 24 to 48 h. P30 tissues were decalcified in 0.5 M EDTA prior to sucrose treatment. Torsos were then mounted in OCT and serially sectioned (12 μm). Next, every fifth section was stained with solution containing 0.5% cresyl violet (Nissl). Cells in both SCGs with characteristic neuronal morphology and visible nucleoli were counted using ImageJ.
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Quantifying Neuronal Populations in Mice
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Other organizations : Johns Hopkins University, Kanagawa Institute of Technology
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Variable analysis
independent variables
- Mouse developmental stage (E16, P0, P30, or P120)
- Neuron counts in the superior cervical ganglia (SCGs)
- Tissue fixation in 4% PFA/PBS for 4 h to overnight
- Cryoprotection in 30% sucrose/PBS for 24 to 48 h
- Decalcification of P30 tissues in 0.5 M EDTA prior to sucrose treatment
- Tissue sectioning (12 μm)
- Nissl staining with 0.5% cresyl violet
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