Fig. 4.

Fig. 4.

Extended applications of the analysis platform including automated analysis of hair cell loss, angiogenesis and eye size in zebrafish embryos. (A) Fluorescent images of YO-PRO-1-stained embryos at 4 dpf, following treatment for 1 h with DMSO, 100 µM pentamidine isethionate (PI) or 100 µM propantheline bromide (PB), analysed for fluorescent granules (red) in the whole fish. (B,C) Total fluorescence intensity in the whole fish (B) and average area of the fluorescent granules (C), showing decrease with drug treatment as previously described (Chiu et al., 2008). (D) Fluorescent images of Tg(kdrl:mCherry) embryos at 2 dpf, treated for 24 h from 24 hpf with DMSO, AG1478 or SU 4312 and analysed for mCherry fluorescence in the whole fish. (E,F) Total fluorescent area in the whole fish following treatment with AG1478 (E) and total fluorescent area in the whole fish following treatment with SU4312 (F), showing reduced angiogenesis in treated fish compared with the control, as previously described (Tran et al., 2007). (G) Brightfield images of mab21l2^+/+, mab21l2^+/− and mab21l2^−/− mutant embryos at 4 dpf analysed for eye size (pink). (H) Eye area, showing the reduced eye size in mab21l2^−/− mutants. Results consistent with previous publications were achieved in a single experiment for each of these assays. Statistical analysis using unpaired t-tests. Error bars show mean±s.d. ns, P>0.05; *P<0.05, ***P<0.001, ****P<0.0001. Scale bars: 500 µm. n refers to the number of embryos analysed for each condition.