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8
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21
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14
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}
],
"issues": [
{
"code": "stale_frames",
"evidence": "In recording 2 from frame 1439 to 2034 (rig clock 312.66s to 332.49s), the camera frame counter stopped incrementing (595 consecutive stale frames with frame counter diff = 0), indicating that the acquisition software re-read the camera buffer.",
"action_taken": "Identified the stale frame interval, invalidated the overlapping stimulation trials (indices 51 to 56) with trial code stale_frames, and excluded the interval from calcium event detection."
},
{
"code": "photobleaching",
"evidence": "Cellular baseline fluorescence faded noticeably across the session, dropping from approximately 1308 counts at the start of recording 1 to 1098 counts at the end of recording 5 (faded to approximately 83% of start, well below the 95% threshold defined in codes.md).",
"action_taken": "Tracked the median relative indicator level across cells throughout the session (which remained above the 50% trial invalidation threshold) and adapted baseline subtraction for calcium event detection."
}
],
"summary": "Analysis of dish 0412-C2 across five recordings (250 commanded stimulations, 30 segmented cells). Key hardware and data issues identified: (1) In recording 2, 595 consecutive stale frames occurred between rig clock 312.66s and 332.49s, invalidating trials 51-56. (2) Noticeable photobleaching occurred across the session, with baseline indicator intensity fading to ~83% of initial levels by recording 5. Out of 250 stimulation trials: 219 were valid, 21 were invalidated due to synchronous network bursts (40-80% of cells firing within 150 ms) occurring within [-0.5s, +0.2s] of pulse command, 6 were invalidated due to stale frames, and 4 trials (indices 93, 100, 108, 116) were invalidated due to aiming off-target (> 10 um from target soma center). Calcium event onsets were detected using neuropil-free soma component reconstruction and frame-difference peak analysis, and evoked responses were determined for all valid trials."
}