Fig. 4: Monitoring change with the TA-GANLive.
a, Step-by-step imaging-assistance pipeline using the TA-GANLive in the live-cell acquisition loop. b, Live-cell imaging of dendritic F-actin before (initial), during (frames 1–15) and after (final) application of a stimulation solution (0 Mg2+/Gly/2.4 mM Ca2+). Shown are the confocal (red, top row), synthetic (purple, middle row) and real (orange, middle row) STED images when acquired, and corresponding segmentation masks for F-actin fibres (magenta, bottom row). The series was chosen as a representative example from a total of 72 series. Colour bars: raw photon counts. c, The DC at each time point measured between the current synthetic image and the last acquired reference STED image for the sequence shown in b. Dark grey points indicate that the last acquired real STED (used as reference) is from a previous time step and light grey points connected with a vertical dashed line indicate that a new STED is acquired at this time step, and the DC is recomputed with this new reference. d, Proportion of dendritic F-actin fibres at each time point segmented by the U-NetLive on either the real STED (orange) or the synthetic STED (purple) images. When a real STED acquisition is triggered, the proportion of fibres in both images is compared (dotted line). Initial and final reference STED images (empty orange circles) are acquired at each round. e, The DC is computed for the F-actin fibre segmentation on control sequences of two consecutive real STED images (time points t and t + 1)). The segmentation of the STEDt image is used as reference and the DC is computed with the segmentation mask on the STEDt+1 image. When a real STED image acquisition would not have been triggered by the threshold-based approach, the DC between the segmentation masks of the two real STED is higher. n = 60 control sequences of two consecutive confocal–STED pairs. Violin plots show the minimum, maximum and mean. Statistical analysis: two-sided Mann–Whitney U test62 for the null hypothesis that the two distributions are the same (***P = 0.0004). Scale bars, 1 μm.
