Protocol for characterization of spatiotemporal network dynamics in cortical and hippocampal assembloids.
The 7 matches
- [1] § Troubleshooting › Potential solution ↔ analyze_calcium_imaging.m, lines 9 to 19 · score 0.76 · temporal smoothing window, max_lag, analyze calcium imaging, frame_period, correlated · Report a mismatch
- [2] § Step-by-step method details › Network analysis ↔ analyze_calcium_imaging.m, lines 197 to 230 · score 0.72 · spectral coherence, analyze calcium imaging, global synchrony, firing rates, burstiness, connectivity · Report a mismatch
- [3] § Step-by-step method details › Network analysis ↔ analyze_calcium_imaging.m, lines 117 to 162 · score 0.69 · functional connectivity matrix, cross correlation, max_lag, ROIs · Report a mismatch
- [4] § Step-by-step method details › Network analysis ↔ example_visualization.m, lines 202 to 231 · score 0.69 · spectral coherence, global synchrony, firing rates, metrics, Network, burstiness · Report a mismatch
- [5] § Step-by-step method details › Network analysis ↔ analyze_calcium_imaging.m, lines 117 to 162 · score 0.65 · R_pvalue, spike train shuffling, n_shuffles, connection · Report a mismatch
- [6] § Step-by-step method details › Network analysis ↔ analyze_calcium_imaging.m, lines 9 to 19 · score 0.53 · max_lag, frame_period, temporal · Report a mismatch
- [7] § Step-by-step method details › Network analysis ↔ analyze_calcium_imaging.m, lines 71 to 105 · score 0.53 · inter spike interval, burstiness, ROI · Report a mismatch
Paper
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The authors' code
MATLAB · 238 lines · 9.2 KB · CC-BY-4.0 · 6 matches
- %% Calcium Imaging Analysis Script
- % This script analyzes processed calcium imaging data from CaImAn/ezcalcium
- % It calculates burstiness, firing rates, spike probabilities, functional connectivity, and synchronization metrics
- %% Clear workspace
- clear;
- clc;
- %% User-defined parameters (modify these as needed)
- Nsamples = 1000; % Number of samples for spike train generation
- frame_period = 0.3875; % Frame period in seconds per frame
- max_lag = 13; % Maximum lag for cross-correlation in frames (~5 seconds at default frame rate)
- % Temporal smoothing parameters
- smooth_window = 1; % Moving average window size for P_spike (1 = no smoothing, >1 = apply smoothing)
- % Statistical significance parameters
- n_shuffles = 1000; % Number of shuffles for significance testing
- significance_alpha = 0.10; % Significance level (e.g., 0.10 for 90% confidence)
- %% Select files to process
- fprintf('\nSelect processed files to analyze...\n');
- [filenames, folderpath] = uigetfile('*.mat', ...
- 'Select .mat files containing refined ROIs to analyze', ...
- 'MultiSelect', 'on');
- if isequal(filenames, 0)
- error('No files selected. Exiting.');
- end
- % Handle single vs multiple file selection
- if ischar(filenames)
- filenames = {filenames}; % Convert to cell array for consistency
- end
- filelist = fullfile(folderpath, filenames);
- fprintf('Selected %d files for processing.\n\n', numel(filelist));
- %% Process each file
- for fi = 1:numel(filelist)
- fil = filelist{fi};
- fprintf('Processing file %d/%d: %s\n', fi, numel(filelist), fil);
- %% Load data
- % Load main variables
- lf = load(fil, 'ROI_center_refined', 'F_raw_refined', ...
- 'F_inferred_refined', 'Z_mod_refined');
- params = struct();
- try
- lf_params = load(fil, 'options');
- params = lf_params.options;
- fprintf(' Loaded existing parameters from file.\n');
- catch
- fprintf(' No parameters found in file. Using defaults.\n');
- end
- params.Nsamples = Nsamples;
- [n_rois, n_frames] = size(lf.Z_mod_refined);
- total_time = n_frames * frame_period;
- fprintf(' Number of ROIs: %d\n', n_rois);
- fprintf(' Number of frames: %d\n', n_frames);
- fprintf(' Total time: %.2f seconds\n', total_time);
- %% Initialize output variables
- burstiness = zeros(n_rois, 1);
- firing_rate_hz = zeros(n_rois, 1);
- P_spike = zeros(n_rois, n_frames);
- %% Process each ROI
- fprintf(' Analyzing ROI spike statistics...\n');
- for i = 1:n_rois
- sinfo = cont_ca_sampler(lf.Z_mod_refined(i, :), params);
- ntrials = size(sinfo.ss, 1);
- all_isi = [];
- total_spikes = 0;
- spike_counts = zeros(1, n_frames);
- for trial = 1:ntrials
- spike_times = sinfo.ss{trial};
- spike_counts = spike_counts + histcounts(spike_times, 0:n_frames);
- if numel(spike_times) > 1
- isi = diff(spike_times) * frame_period;
- all_isi = [all_isi; isi(:)];
- end
- total_spikes = total_spikes + numel(spike_times);
- end
- % Burstiness: CV of inter-spike intervals
- if ~isempty(all_isi) && std(all_isi) > 0
- burstiness(i) = std(all_isi) / mean(all_isi);
- else
- burstiness(i) = 0;
- end
- firing_rate_hz(i) = total_spikes / (ntrials * total_time);
- P_spike(i, :) = spike_counts / ntrials;
- end
- fprintf(' Burstiness range: [%.3f, %.3f]\n', min(burstiness), max(burstiness));
- fprintf(' Firing rate range: [%.3f, %.3f] Hz\n', min(firing_rate_hz), max(firing_rate_hz));
- %% Apply temporal smoothing to P_spike
- if smooth_window > 1
- fprintf(' Applying temporal smoothing (window size = %d frames)...\n', smooth_window);
- P_spike_smoothed = zeros(size(P_spike));
- for i = 1:n_rois
- P_spike_smoothed(i, :) = movmean(P_spike(i, :), smooth_window);
- end
- P_spike = P_spike_smoothed;
- end
- %% Calculate functional connectivity using cross-correlation
- fprintf(' Computing functional connectivity matrix...\n');
- R = zeros(n_rois, n_rois);
- R_pvalue = ones(n_rois, n_rois);
- R_significant = false(n_rois, n_rois);
- fprintf(' Computing significance thresholds via spike train shuffling (%d shuffles)...\n', n_shuffles);
- for i = 1:n_rois
- for j = i:n_rois
- if i == j
- R(i, j) = 1.0;
- R_pvalue(i, j) = 0;
- R_significant(i, j) = true;
- else
- [xcorr_vals, ~] = xcorr(P_spike(i, :), P_spike(j, :), max_lag, 'coeff');
- % Store signed max correlation (preserves excitatory/inhibitory)
- [~, max_idx] = max(abs(xcorr_vals));
- actual_corr = xcorr_vals(max_idx);
- R(i, j) = actual_corr;
- R(j, i) = actual_corr;
- % Null distribution via circular shift shuffling
- null_corr = zeros(n_shuffles, 1);
- for shuffle = 1:n_shuffles
- random_shift = randi([1, n_frames - 1]);
- P_shuffled = circshift(P_spike(j, :), random_shift);
- xcorr_jitter = xcorr(P_spike(i, :), P_shuffled, max_lag, 'coeff');
- [~, max_idx_jitter] = max(abs(xcorr_jitter));
- null_corr(shuffle) = xcorr_jitter(max_idx_jitter);
- end
- % Two-tailed test
- p_val = sum(abs(null_corr) >= abs(actual_corr)) / n_shuffles;
- R_pvalue(i, j) = p_val;
- R_pvalue(j, i) = p_val;
- is_significant = p_val < significance_alpha;
- R_significant(i, j) = is_significant;
- R_significant(j, i) = is_significant;
- end
- end
- end
- %% Calculate synchronization metrics
- upper_triangle_indices = triu(true(n_rois), 1);
- unique_correlations = R(upper_triangle_indices);
- n_significant = sum(R_significant(upper_triangle_indices));
- percent_significant = 100 * n_significant / numel(unique_correlations);
- % Global population synchrony (z-score normalized, all neurons)
- fprintf(' Computing global population synchrony...\n');
- P_spike_norm = zscore(P_spike, 0, 2);
- global_synchrony_timeseries = 1 ./ (std(P_spike_norm, 0, 1) + eps);
- global_synchrony_index = mean(global_synchrony_timeseries);
- % Spectral coherence (frequency-domain, all pairs)
- fprintf(' Computing spectral coherence...\n');
- coherence_matrix = zeros(n_rois, n_rois);
- for i = 1:n_rois
- coherence_matrix(i, i) = 1.0;
- for j = i+1:n_rois
- [Cxy, ~] = mscohere(P_spike(i, :), P_spike(j, :), [], [], [], 1/frame_period);
- avg_coherence = mean(Cxy);
- coherence_matrix(i, j) = avg_coherence;
- coherence_matrix(j, i) = avg_coherence;
- end
- end
- spectral_coherence_index = mean(coherence_matrix(upper_triangle_indices));
- fprintf(' Global synchrony index: %.4f\n', global_synchrony_index);
- fprintf(' Spectral coherence index: %.4f\n', spectral_coherence_index);
- fprintf(' Significant connections: %d/%d (%.1f%%)\n', ...
- n_significant, numel(unique_correlations), percent_significant);
- fprintf(' Functional connectivity range: [%.3f, %.3f]\n', ...
- min(unique_correlations), max(unique_correlations));
- %% Save results
- fprintf(' Saving results...\n');
- original_data = load(fil);
- original_data.burstiness = burstiness;
- original_data.firing_rate_hz = firing_rate_hz;
- original_data.P_spike = P_spike;
- original_data.R = R;
- original_data.R_pvalue = R_pvalue;
- original_data.R_significant = R_significant;
- original_data.coherence_matrix = coherence_matrix;
- original_data.global_synchrony_timeseries = global_synchrony_timeseries;
- original_data.global_synchrony_index = global_synchrony_index;
- original_data.spectral_coherence_index = spectral_coherence_index;
- original_data.params = params;
- original_data.analysis_metadata = struct(...
- 'frame_period', frame_period, ...
- 'max_lag', max_lag, ...
- 'Nsamples', Nsamples, ...
- 'smooth_window', smooth_window, ...
- 'n_shuffles', n_shuffles, ...
- 'significance_alpha', significance_alpha, ...
- 'n_rois', n_rois, ...
- 'n_frames', n_frames, ...
- 'total_time', total_time, ...
- 'n_significant_connections', n_significant, ...
- 'percent_significant_connections', percent_significant, ...
- 'analysis_date', datetime('now'));
- save(fil, '-struct', 'original_data');
- fprintf(' File updated successfully.\n\n');
- end
- %% Summary
- fprintf('========================================\n');
- fprintf('Analysis complete!\n');
- fprintf('Processed %d files.\n', numel(filelist));
- fprintf('========================================\n');
analyze_calcium_imaging.m, under CC-BY-4.0 · at the source
Overview
- Department of Neurology, University of California, Los Angeles, David Geffen School of Medicine, Los Angeles, CA 90095, USA
- Department of Neurobiology, University of California, Los Angeles, Los Angeles, CA 90095, USA
- Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA 90095, USA
- Intellectual Development and Disabilities Research Center, University of California, Los Angeles, Los Angeles, CA 90095, USA
- Department of Anatomy and Neurobiology, School of Medicine, University of California, Irvine, Irvine, CA, USA
- Sue & Bill Gross Stem Cell Research Center, School of Medicine, University of California, Irvine, Irvine, CA, USA
Abstract
This protocol generates neural assembloids from human induced pluripotent stem cells (hiPSCs) to model human brain circuitry. It details the differentiation of excitatory-predominant hippocampal (Hc) and cortical (Cx) organoids and their fusion with inhibitory interneuron-predominant ganglionic eminence (GE) organoids. We then detail procedures for maintaining assembloids to promote interneuron migration and network integration, followed by functional assessment using two-photon calcium imaging to measure neuronal activity.
For complete details on the use and execution of this protocol, please refer to McCrimmon et al.1
Reproduced under the paper's license (CC BY), from the paper cited above.
Repository
Its files are read in the Code ↔ Paper reader above, with 7 matches between paragraphs and lines of code.
Zenodo 18763819
Availability: 1 check, the latest on 30 September 2026: the link answers (HTTP 200)
- 30 September 2026: the link answers (HTTP 200)
2 files
- analyze_calcium_imaging.
m , MATLAB, 238 lines, 6 matches - example_visualization.m, MATLAB, 231 lines, 1 match
The paper's code and data availability statement is in the Data section.
Tracing map
Proposed by the machine: these links were found in the paper and verified at the source, without human review. The map will receive a Zenodo DOI once one of the paper's authors has validated it with their ORCID.
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- 7 matches between paragraphs of the paper and lines of the code (method lexical-v1);
- neither the text of the paper nor the code itself.
Its JSON (tracing-map.json) is deposited on Zenodo with its DOI once the map is validated.
Data
Datasets cited
- github.com/
samarasinghelab/ , at github.com; found in the text, “Processing using EZcalcium in MATLAB”star_protoc
Data and code availability
The accession number for the original MATLAB code (analyze_calcium_imaging
Reproduced under the paper's license (CC BY), from the paper cited above.
Versions
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Version 1, 30 September 2026: the first record
Recorded: type, language, journal, volume, issue, pages, dates, 6 authors, 3 keywords, 9 MeSH terms, 5 funders, 13 references, 5 RRIDs.
Cite
This paper
Guzman, A. R., McCrimmon, C. M., Toker, D., Novitch, B. G., Watanabe, M., & Samarasinghe, R. A. (2026). Protocol for characterization of spatiotemporal network dynamics in cortical and hippocampal assembloids. STAR protocols, 7(2), 104463. https://
BibTeX
@article{guzman2026proto
author = {Guzman, Asia R and McCrimmon, Colin M and Toker, Daniel and Novitch, Bennett G and Watanabe, Momoko and Samarasinghe, Ranmal A},
title = {{Protocol for characterization of spatiotemporal network dynamics in cortical and hippocampal assembloids}},
journal = {STAR protocols},
year = {2026},
month = mar,
volume = {7},
number = {2},
pages = {104463},
publisher = {Elsevier},
issn = {2666-1667},
doi = {10.1016/
url = {https://
pmid = {41904955},
pmcid = {PMC13054417}
}
RIS
TY - JOUR
AU - Guzman, Asia R
AU - McCrimmon, Colin M
AU - Toker, Daniel
AU - Novitch, Bennett G
AU - Watanabe, Momoko
AU - Samarasinghe, Ranmal A
TI - Protocol for characterization of spatiotemporal network dynamics in cortical and hippocampal assembloids
T2 - STAR protocols
J2 - STAR Protoc
PY - 2026
DA - 2026/
VL - 7
IS - 2
SP - 104463
SN - 2666-1667
PB - Elsevier
DO - 10.1016/
UR - https://
LA - en
ER -
CSL-JSON
{
"id": "10.1016/
"type": "article-journal",
"title": "Protocol for characterization of spatiotemporal network dynamics in cortical and hippocampal assembloids",
"container-title": "STAR protocols",
"author": [
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"family": "Guzman",
"given": "Asia R"
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{
"family": "McCrimmon",
"given": "Colin M"
},
{
"family": "Toker",
"given": "Daniel"
},
{
"family": "Novitch",
"given": "Bennett G"
},
{
"family": "Watanabe",
"given": "Momoko"
},
{
"family": "Samarasinghe",
"given": "Ranmal A"
}
],
"container-title-short":
"volume": "7",
"issue": "2",
"page": "104463",
"DOI": "10.1016/
"PMID": "41904955",
"PMCID": "PMC13054417",
"ISSN": "2666-1667",
"publisher": "Elsevier",
"URL": "https://
"language": "en",
"issued": {
"date-parts": [
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2026,
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]
}
}
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