Fig. 8

Device-dissociated neurons extend larger arbors and manifest the CASK-LOF bushy phenotype in vitro. Device dissociations were performed with flow parameters 50 µl/sec, infusion volume 10.5 µl, 4.8 Hz, mean cycle number1444. Neurite-arbor size parameters from 3 div cultures, comparing methods of dissociation and genotypes. Box-plot distributions depicted as in Fig. 3. Significance levels: *, p < 0.05; **, p < 0.005; ***, p < 0.0005; ****, p < 0.00005; ns, not significant. a-f Device vs. manual dissociation. a-c Ex33/Ex33. Device dimensions: channel height 500 µm; orifice length 400 µm; orifice width 60 µm. d-f ∆18/∆18. Device dimensions: channel height 450 µm; orifice length 400 µm; orifice width 50 µm. For both genotypes, total neurite length (a, d), territory area (b, e), and higher-order branch number (c-f) were significantly higher for microfluidic device-dissociated neurons. g-j Comparison of device-dissociated neurons from control (Ex33/Ex33; one sample) and CASK mutant (∆18/∆18; two samples) larval CNS. Device dimensions: channel height 450 µm; orifice length 400 µm; orifice width 50 µl. g Total neurite length. h Territory area. i Higher-order branches. j Branch density. Device-dissociated CASK-mutant neurons extend bushy neurite arbors, replicating the phenotype seen in manual cultures. In addition, two cultures prepared after serial device-based dissociation show marked consistency of neurite-arbor parameters