Fig. 8.

Calcofluor staining of mutant and wild-type pollen tubes. Quartets were cultured in vitro at 16 °C for 12 h and then used for S4B staining. (A–H) Spinning disk confocal and bright field images of wild-type qrt1/qrt1 (A, B), csld4-3/+; qrt1/qrt1 (C, D), csld1-1/+; qrt1/qrt1 (E, F), and csld1-1/+; csld4-3/+; qrt1/qrt1 (G, H) pollen tubes stained with Calcofluor. Arrows indicate abnormal mutant pollen tubes. (I) and (J) Calcofluor staining of the tip region of a normal pollen tube, which cannot be shown in the same images above. (K) The fluorescence intensity of mutant pollen tubes was much weaker than that of control tubes. Images were taken at the same focal plane and the fluorescence intensity along the pollen tube wall was quantified by the grey value measured with ImageJ (Abramoff et al., 2004; http://rsbweb.nih.gov/ij/). The fluorescence intensity of the brightest pollen tube was set to 100% and the weakest was measured (n=20 for wild-type qrt1/qrt1, n=26 for csld1-1, n=24 for csld4-3, and n=18 for the csld1 csld4 double mutant). Bars: 20 μm.