Fig. 3: High-fidelity voltage imaging enhancement via FAST.
From: Real-time self-supervised denoising for high-speed fluorescence neural imaging
a Workflow for simulating two-photon voltage imaging data. Ground truth (GT) signals with varying spike widths (2 ms, 4 ms, 6 ms, or 8 ms), were combined with mixed Poisson-Gaussian noise to generate network inputs. b Statistical analysis of denoising performance across spike widths. Pearson correlation coefficients were calculated between each neuron’s denoised signals (five methods) and the GT signal. Data are presented as violin plots (n = 10 neurons). Statistical significance was determined using two-sided two-way ANOVA followed by Tukey’s multiple comparisons test. Significance levels are indicated as follows: NS (P ≥ 0.05); ** (P < 0.01, decrease); *** (P < 0.001, decrease); ## (P < 0.01, increase); ### (P < 0.001, increase). c Representative ΔF/F traces from a single simulated neuron, comparing GT, noisy, and denoised signals across the four spike widths. d Raw and FAST-denoised images with in vivo simultaneous voltage imaging and electrophysiology. Left: Raw and FAST-denoised images of a QuasAr6a-expressing neuron in mouse cortex (L2/3) at 1000 Hz (data from ref. 53). Right: Electrophysiology trace (top) and ΔF/F traces from 1000 pixels within the neuron ROI (annotated using the ROI Manager toolbox in Fiji54) for noisy (middle) and FAST-denoised (bottom) data. Red box is magnified in (e). Scale bar, 10 μm. Representative frames are shown in the figure, and similar results were obtained across n = 4982 frames in all experiments. e Enlarged view from (d). Overlaid raw and FAST-denoised ΔF/F traces from three representative pixels are aligned with the electrophysiology trace. Red triangles mark spike peaks. f Pearson correlation between the n = 100 single-pixel ΔF/F traces and the electrophysiology recording in (d). FAST significantly improves correlation (***P < 0.001, unpaired two-tailed t-test). g Voltage imaging of zebrafish spinal cord neurons using light sheet microscopy. Raw and FAST-denoised images of zArchon1-expressing neurons imaged at 1000 Hz (data from ref. 16.). ROIs for three representative cells are manually annotated and magnified on the right. Scale bars, 20 μm (left), 5 μm (right). h ΔF/F traces for the three neurons from g, comparing raw and FAST-denoised signals. Red box is magnified in (i). i Enlarged view from (h), with raw and FAST-denoised traces overlaid for direct comparison.
