Fig. 3: Dual-color volumetric imaging of live RBL cells.

a–d Dual-color volumetric imaging of granule motions in a live RBL-2H3 GFP-FasL cell, where the cell membrane is labeled with IgE-CF640R and granules contain GFP-FasL, at an imaging speed of ~ 0.6 volumes (80 × 15 × 40 µm³ in XYZ) per second for 80 volumes, for a total imaging time of ~2 min. a Maximum intensity projection views of the cell images at one-time point and overlay with representative trajectories of granule movement (orange lines). b Time series of the trajectories in a. c, d Histograms of estimated diffusion coefficients and velocities of all trajectories (n = 185) found in cell 1 and cell 2. e–g Dual-color volumetric imaging of live RBL-2H3 GFP-FasL cell, where the cell membrane is labeled with CellMask DeepRed and the granules contain GFP-FasL using an imaging speed of ~8.3 volumes/s for 80 volumes for a total time of 10 s. e Maximum intensity projection views of the cell images at one-time point and overlay with representative trajectories of granule movement (orange lines). f Time series of the trajectories in e. To indicate the current positions of the tracked particles, we used lighter orange lines represent past trajectories. g Histograms of estimated diffusion coefficients of all trajectories (n = 48) in the cell. h Cumulative probability of the estimated diffusion coefficients under normal (a–d) and stressed (e–g) imaging conditions. 400–500 trajectories with a diffusion coefficient >0.001 µm²/s from four cells under each condition are selected.