Figure 4

Simultaneous tricolor AFM-LSCM images of C. albicans showing the localization of tubulin2-GFP (Tub2-GFP, green), histone protein B-RFP (Htb-RFP, purple) and onset of ROS (Cell-ROX-deep red, red) after 2,4-D exposure. (A–D) Show multicolored LSCM images overlaid onto the AFM QI image before (A,B) and after (C,D) 2,4-D exposure. Confocal images were taken at mid-cell, whereas AFM quantitative images were at the surface, making the two images look very different. (E–H) Show control images taken before the addition of 8 mM 2,4-D, indicating the absence of a ROS signal (H). (I–L) Are images collected 5 min after the addition of 2,4-D, clearly showing a ROS signal (L). (E and I) Show tricolour merged images with Tub2-GFP, Htb-RFP and Cell-ROX, F and J show only RFP, G and K show only GFP and H and L show only ROS.