Fig. 4: SFSRM enables noninvasive super-resolution imaging in live cells at millisecond temporal resolution for thousands of frames.
From: Single-frame deep-learning super-resolution microscopy for intracellular dynamics imaging
a Representative WF and SFSRM images of endoplasmic reticulum in live Beas2B cells transfected with EGFP-Sec61β. b Time-lapse images of endoplasmic reticulum in live cells imaged with 15 W/cm2 intensity illumination for 5000 frames. First row: representative raw low-SNR images. Second row: the corresponding SFSRM reconstructions from the low-SNR images. Third row: Error maps of SFSRM reconstructions compared to the raw low-SNR images. The reconstruction errors are analyzed by SQUIRREL analysis, which gives an error map to indicate the possible local errors. Fourth row: disagreement maps measured by ensemble disagreement method, in which the regions with high disagreement score could be less trustworthy. c Representative WF and SFSRM images of mitochondria in live Beas2B cells transfected with Tom20-mcherry. d Comparison of the WF and corresponding SFSRM time-lapse images showing a mitochondrial “kiss-and-run” process. e Comparison of the mitochondrial segmentation results from the WF and SFSRM images. The target mitochondria in the segmentation results are marked in magenta while the other mitochondria are marked in white. f Comparison of the counted transient fusion and fission rates in the SFSRM and WF sequences. Boxplots are drawn from the 25th to 75th percentile with the horizontal bar at the median and the whiskers extending to the minima and maxima. Fusion and fission rates in 10 s were analyzed. g Comparison of measured mitochondrial area variation of the target mitochondrion in the SFSRM and WF sequences. h Representative WF and SFSRM images of microtubules in Beas2B cells expressing mEmerald-ensconsin. i Microtubule bundle instability caused by inconsonant fluctuations of microtubules. j Comparison of the WF and SFSRM images of the microtubules. The temporal-color-coded images, which are the maximum projected images of the time-lapse WF/SFSRM images with each frame rendered with different colors, indicate the microtubule transverse movement over time. k The transverse positions of a single microtubule over time recorded at 20 Hz. l Histogram of microtubule transverse displacement at a 50-ms interval. Scale bar, 2 μm (a–c, e, h), 500 nm (d, i, j).
