Fig. 3. The cloche lens has cataracts. (A) The morphology of the eye was analyzed by bright-field microscopy at 2.5 dpf. (B) Sections were prepared from 2-4 dpf embryos and analyzed by Hematoxylin and Eosin. The green arrow (3 dpf) and red arrow (4 dpf) point to nuclei. (C) The number of lens fiber cell nuclei per section was analyzed at 2-4 dpf. Three sections of each lens were analyzed by an image analysis program (ImageJ). Fourteen embryos for each time point were analyzed. The mean±s.d. is shown. (D) The reflected light (reflectance) of the lens was detected by confocal microscope at 2-4 dpf. The red arrows point to areas of reflected light. (E) Statistical analysis of reflectance intensity. The Mann-Whitney U test was used to test for differences between two groups. The red line in the box corresponds to the median value. The differences between wild-type and cloche^m39 at 3 dpf (P=0.0339) and 4 dpf (P=0.0209) are significant. (F) The reflected light (reflectance) of the lens in cloche^m39 (clo^m39) and cloche^s5 (clo^s5) was compared by confocal microscopy at 4 dpf. Reflectance is comparable between the two. The red arrows indicate areas of reflected light. (G) Statistical analysis of reflectance intensity. The Mann-Whitney U test was used to test for differences between two groups. The red line in the box corresponds to the median value. The differences between wild type and cloche^m39 (clo^m39) (P=0.0209) or cloche^s5 (clo^s5) (P=0.0143) are significant.