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. Author manuscript; available in PMC: 2026 Jun 29.
Published in final edited form as: Nat Methods. 2026 May 22;23(6):1184–1195. doi: 10.1038/s41592-026-03066-1

Fig. 2 ∣. Multimodal super-resolution imaging of subcellular dynamics.

Fig. 2 ∣

a, Nuclear imaging (SPY 505) and single-particle tracking with LLSM, visualizing molecular trajectories and diffusion dynamics of SOX2 (HaloTag-PA-JF646) in mouse embryonic stem (mES) cells at 50 Hz. Scale bar (left), 5 μm. Scale bar (top right), 2 μm. The probability density histogram of log10 diffusion coefficients is binned from −3 to +3 in 0.1-unit increments. b, Top: LLS-SIM xy MIP of hTERT-RPE1 cells expressing ER (StayGold-ER, gray) and Golgi (β4Gal-HaloTag9 labeled with JFX549, orange) markers, from Supplementary Video 6. Insets, corresponding xy and yz OTFs. Scale bar, 25 μm. Bottom, volume renderings from boxed regions above showing ER and Golgi organization. Scale bar, 2 μm. c, Comparison of widefield and 3D-SIM imaging in hTERT-RPE1 cells with mitochondria (COX8a-StayGold, cyan) and Golgi marker (orange). Scale bar, 25 μm. Insets, 3D-SIM OTFs. d, Timelapse 3D-SIM (Supplementary Video 7) captures mitochondrial and Golgi dynamics during cell division (white box from c). Scale bar, 5 μm. e, Correlative optical microscopy applying multiple imaging modalities (widefield, 3D-SIM, OI, LLS and LLS-SIM) to a dividing hTERT-RPE1 cell with labeled mitochondria (blue–green) and Golgi (orange), from Supplementary Video 8. Scale bar, 10 μm. f, Timelapse sequence of the correlative imaging shown in e. Scale bar, 10 μm.