Fig. 4: Imaging of peroxisomes and ER in live cells at a 100 volumes per second (vps) using Alpha-LFM. | Nature Communications

Fig. 4: Imaging of peroxisomes and ER in live cells at a 100 volumes per second (vps) using Alpha-LFM.

From: Adaptive-learning physics-assisted light-field microscopy enables day-long and millisecond-scale super-resolution imaging of 3D subcellular dynamics

Fig. 4

a Volume rendering of peroxisomes (tagged with SKL-mApple) in a live U2OS cell, acquired with Alpha-LFM. b Time-lapse MIPs of the ROI indicated by orange boxes in (a). Arrows indicate the rapid motion of peroxisomes. c 3D tracking of peroxisomes imaged at 100 vps and those downsampled to 10 vps, with velocity encoded by color. d Velocity plots of the peroxisome highlighted by a box in (c). e Trajectory of the peroxisome highlighted by box in (c). f Comparison of trajectories tracked using the 100-vps and 10-vps results. g x-y MIP and x-z, y-z slices of the ER (tagged with Sec61β-EGFP) in a live COS-7 cell captured by Alpha-LFM. h Projection of skeletonized images over 1 s, with time encoded by color, visualizing ER dynamics. Scale bar, 5 μm. i Comparison of ER dynamics of the ROI (marked by white boxes in g, h) acquired at 100 vps versus 1 vps (downsampled). White arrows and circles highlight dynamics resolved at 100 vps but blurred at 1 vps. Scale bar, 2 μm. j, k Magnified time-lapse images of the ROI marked by the orange box in (g). White arrows in (j) indicate the stretching of an ER tubule within 40 ms, while orange arrows in (k) show the formation of a new tubule within 40 ms. Scale bar, 2 μm.

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