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26
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24
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14,
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13
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18
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8,
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0,
18
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{
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{
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29
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8
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3
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{
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21
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{
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14
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{
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0
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],
"issues": [
{
"code": "stale_frames",
"evidence": "Recording 2: 595 consecutive saved frames (rig clock 312.66-332.49 s) all carried camera frame counter 5222 and were identical (frame mean and every per-cell value constant), i.e. the camera delivered no new frames and the acquisition software re-read its buffer.",
"action_taken": "The frozen stretch was flagged; stimulations whose -0.25 s/+0.4 s window touched it (indices 51-56) were marked invalid with stale_frames, and no events were called from the frozen frames."
},
{
"code": "photobleaching",
"evidence": "Median over cells of the soma (ROI) signal fell steadily from 1.00 at the start of recording 1 to 0.84 by the end of recording 5 (0.96, 0.90, 0.86, 0.84 at the ends of recordings 1-5), well below the 95% mark.",
"action_taken": "Bleaching was tracked and used as the running baseline for the dF/F; it never reached the below-half level, so no trial was invalidated for it."
}
],
"summary": "Five recordings of dish 0412-C2 (30 segmented cells) with 250 logged single-cell red stimulations. Events were extracted per cell by matched-filtering the neuropil-subtracted weighted-ROI dF/F with a per-cell calcium kernel, refining onsets with a deconvolution step and grouping spikes closer than 0.3 s; about 1086 events were found. Only a subset of cells expresses the opsin: some targets never responded, others responded almost every time, and a typical stimulation recruited 0-3 cells. 25 network bursts (12-25 of 30 cells firing within 150 ms) occurred, most starting 30-100 ms after a stimulation; 21 stimulations fell inside a burst window and were marked invalid. Recording 2 lost 19.8 s of data (595 re-read frames, 312.7-332.5 s), invalidating stimulations 51-56. Four stimulations were aimed 10-21 um off their target (indices 93, 100, 108, 116). The indicator bleached to 84% of its starting level over the session. No dropped frames, room light or camera-link loss were seen."
}