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. 2012 Oct 24;7(10):e47552. doi: 10.1371/journal.pone.0047552

Figure 1. Generation of axoplasm-enriched optic nerve samples using a detergent-free method.

Figure 1

(1) Dissected optic nerve is cut into ≈1 mm segments and positioned so that axon orientation is at 90 degrees to the pHEMA disk; optic nerve segments are incubated on the pHEMA disks for 1 hr at 4C. (2) Tissue is removed and two pHEMA disks are placed into a QIAshredder tube, overlaid with 80 ul of axoplasm buffer (0.5 M TEAB + protease inhibitor) and incubated at 4C for 10 min. Tubes are then spun at 12,000 g for 10 min, followed by the addition of a further 50 ul of axoplasm buffer, a 5 min incubation at 4C and a further spin at 12,000 g. (3) Axoplasm-enriched samples are collected and placed at −20C for storage.