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Fig. S4 Confocal imaging does not notably inhibit fungal growth, filamentation, or virulence. (A–C) CAF2-dTomato C. albicans were microinjected into the hindbrain ventricle of AB zebrafish and fish were screened for inoculum. One group of fish was imaged from 0 to 9 hpi, while the other was imaged only at 9 hpi. Imaging performed with 543 nm He-Ne laser at 88% power, using a 40× 0.75NA objective, with slices at 1.5 μm spacing. Pixel dwell time was 4 μs, each image dimension was 1024×1024, and total number of slices ranged from 32–40, with total active imaging time of 5–7 minutes per stack and 45–63 minutes for each fish total over the course of the experiment. (A) Image panels of a fish imaged from 0 to 9 hpi with florid fungal filamentation. Scale bar = 10 μm. (B) The number of C. albicans cells per fish was quantified at 9 hpi for each imaging group (N = 3) with no statistical significance (p>0.05). (C) The number of C. albicans filaments per fish were quantified at 9 hpi for each imaging group (n = 3) with no statistical significance (p>0.05). Error bars represent standard error of the mean. These results are representative of a large number of experiments that have been performed but were not specifically quantified. (D) Comparison of mortality in photoswitching experiments. Fish were infected and imaged as described for Fig. 1. Photoswitching was carried out as described in the Materials and Methods. Mortality was assayed at 24 hpi for photoswitched Tg(mpeg1:GAL4/UAS:Kaede) fish (N = 12) and non-photoswitched Tg(mpeg1:GAL4/UAS:Kaede) fish (N = 8). This is compared to typical 24 hpi mortality (35%) seen in this infection model for control fish in experiments described in Fig. 5 and Fig. 6 (n = 8 independent experiments).