Journal: bioRxiv
Article Title: Spike inference from calcium imaging data acquired with GCaMP8 indicators
doi: 10.1101/2025.03.03.641129
Figure Lengend Snippet: To evaluate the transfer function of various CASCADE models and their nonlinearity, a synthetic ground truth was generated. The ground truth spike patterns consisted of experimentally recorded action potentials of excitatory neurons from refs. , , , (n = 237 neurons with variable recording duration, each represented by a gray line; 102,093 action potentials, 32.7 hours of recordings) to ensure naturalistic spike patterns. These experimentally obtained spike patterns were convolved with a double exponential kernel (rise time 5 ms, decay time 500 ms, peak amplitude 40% ΔF/F), with Gaussian noise added to reach a standardized noise level of “8”. Spike inference algorithms were applied to this entire linear synthetic dataset. From the inferred spike rates and the synthetic ground truth, transfer functions as in were retrieved, allowing to judge the linearity of the models trained on specific datasets. The Default CASCADE model exhibited the highest nonlinearity (black average transfer curve) compared to the weighted linear fit of the inferred spike rates (red dashed line); the sublinearity of the average transfer function at high firing rates reflects the supralinearity of the training data. CASCADE trained with GCaMP8 ground truth yielded a relatively linear transfer function, with mild but clear signs of saturation for models fine-tuned for GCaMP8f and GCaMP8s. The transfer function obtained from a model trained with GCaMP8m (GC8m-tuned CASCADE) data was the most linear. Since the models reflect the nonlinearities of their training data, these analyses indicate that GCaMP8, and in particular GCaMP8m, are distinctly more linear calcium indicators than for example GCaMP6 (which is the basis of most of the training data for Default CASCADE) across the firing rate regime that is typically covered by experimentally obtained ground truth recordings. The bottom right panel shows quantified deviations from linearity as introduced in for the different CASCADE models. Quantifications were pooled across multiple simulations with variable setting of the kernel parameters to emulate GCaMP8f, GCaMP8m, GCaMP8s, GCaMP7f and GCaMP6 with the kernel rise times (2, 3, 4, 11 and 50 ms, respectively), decay times (45, 80, 190, 100 and 300 ms) and peak amplitudes (55, 71, 81, 70 and 25 % ΔF/F). Deviations from linearity were reduced compared to Default CASCADE by 30% (GC8-trained), 19% (GC8f-trained), 60% (GC8m-trained) and 48% (GC8s-trained). Therefore, the GC8m-trained model exhibited the lowest deviation from linearity, suggesting the highest linearity of the GC8m indicator used for its training. All comparisons p ≪ 10 −10 , Wilcoxon signed-rank test, compared across n = 1422 instances of simulated neurons.
Article Snippet: A modified Gaussian mixture model (GMM) in MATLAB was then used to perform a quantal analysis from the histogram to infer the unitary amplitude.
Techniques: Generated