Figure 1.

Details of calibration of IRM. (a) Given here is an IRM setup schematic showing illumination and detection path. Zoomed-in view shows reflections at interface 1 (coverslip-medium, blue arrows) and 2 (medium-cell, green arrows), which interfere. (b) (Top) Shown here is an IRM image of a bead with a typical radial line in yellow. (Bottom) Shown here is an averaged intensity-versus-height profile (N = 10) with linear fit of the first branch in red. (c) (Top) Shown here is a plot of I_max, I_min and S/2 from profiles of same beads is imaged at different exposure times (n = 5 beads, four line scans per bead) for a particular day. (Bottom) Given here is ΔI/Δh versus exposure times for beads. (d) Shown here is a typical IRM image of a HeLa cell. White ROIs are used to calculate S/2 (cell). (e) Given here is a comparison of I_max, I_min and S/2 between beads and cells for different days (N = 20). Gray region covers y = x ± 0.1 x. (f) Shown here is the SD_(time) measured and the S/2 (cell) used for different days versus the conversion (ΔI/Δh) used. (g) Shown here are minima (red) and maxima (green) projections of an HeLa cell with FBRs overlaid in cyan. (h) Shown here, FBRs are overlaid on the corresponding IRM image. (i) Shown here are relative height maps at any given time-point of six FBRs in an HeLa cell. Scale bars represent 1 μm. Scale bars for (a–h) = 10 μm. See also Fig. S1; Supporting Discussion. To see this figure in color, go online.