β cell death and CTL motility after antigen depletion.
(A–C) Events of β cell death with loss of GFP as a readout measure (red arrows). A corresponds to Supplemental Video 14. B is derived from the same sequence used in Figure 5C. Imaging volume V and number of unique CTL tracks are indicated. Dimensions: A, w/h = 0.58/d = 6/z = 21; B, w/h = 1/d = 6/z = 22; C, w/h = 1.08/d = 6/z = 10; t = 30 s for all sequences. (D) Representative control islet from naive animal imaged at maximum laser intensity; w/h = 0.51/d = 5/z = 22. (E) Association of arrested CTL (CFP, pseudocolored magenta; white arrows) with dying β cells (GFP, green; red arrows). E corresponds to Supplemental Videos 15 and 16; w/h = 0.86/d = 5/z = 22. (F–I) Representative images from full-blown diabetic mice and corresponding tracking results. Imaging volume V and number of unique CTL tracks are indicated. Supplemental Video 17 shows an enlarged region of F, while G corresponds to Supplemental Video 18, which includes vascular staining. Representative of 3 individual experiments. Dimensions: F, w/h = 1.52/d = 5/z = 20; G, w/h = 1.52/d = 5/z = 18. Yellow arrows in F and G indicate remaining β cells. P values were obtained with Mann-Whitney analysis. Horizontal bars are means and associated error bars represent the SEM. Scale bars: 100 μm (B, C, E–G); 50 μm (A and D).