Supplementary Figure 15: Motion blur in traditional TIRF-SIM.
From: Single-shot super-resolution total internal reflection fluorescence microscopy
EGFP-Rab11 particles in U2OS cells were imaged at 37 °C on a commercial TIRF SIM system (9 diffraction-limited images acquired and processed to reconstruct a single super-resolution image) at a frame rate of 12.9 Hz. a) Single raw frame from diffraction-limited series (time point 62 of 200). b) Corresponding TIRF SIM reconstruction. c, d) Higher magnification images of the yellow dashed rectangular regions in a, b. See also Supplementary Video 17. Motion blur broadens particle extent significantly (compare particles highlighted with red arrows, apparent particle width given in red text), presumably due to particle motion during the 9-frame raw image acquisition period (Supplementary Video 18). Scale bars in a, b 5 μm; in c, d 500 nm. Experiments were repeated at least 3 times with similar results; representative data are shown.
