Experiment Session Notes

• Mouse ID: 801381
• Date: 2025-05-29
• Experimenter: @Seyedolmohadesin @rcpeene @jeromelecoq Kaspar
• Rig / Setup ID: SLAP2
• Stimulus version: https://github.com/AllenNeuralDynamics/openscope-community-predictive-processing/blob/3338a178cba99f34894657dad7eaaf872bdbe896/code/stimulus-control/src/Standard_oddball_slap2.bonsai
• Protocol followed: Glutamate (Green fluorophore) + Calcium (Rcamp3 red fluorophore) imaging, 50 min experiment

• Notes & Issues:

  • Similar experiment as last time. This time we record behavior videos (eye, face) with HARP timestamps aligned with the other events. Body camera was turned off as we are having file closing issues for all 3 cameras at the same time. We will fix later on.
  • SLAP2 was ran in CONTINUOUS mode, this means we now have a continuous recording, no more SLAP trials.

  • Screen is now Gamma calibrated. It was NOT before.

  • Baseline: 2405 -> NbBaselineGrating * (1 * (0.343+0.343)) -> 27.49 min

  • Mismatch: 40 -> NbMismatchPerCondition * 4 * (0.343+0.343)) -> 1.829 min of oddballs
  • Mismatch during orientation tuning: 40 -> 14.63 min
  • Receptive field: 10 -> NbReceptiveFieldRepeats * ((0.25) * 149) -> 6.2 min of RF

  Total Expected duration = 50.16 min Oddball occurs on average every 11s

  • Head fixation started at 14:34

Mouse is labelled with GluSNF4 and Rcamp3 both with viruses injection. Injection is rather dense. with both color present. We will image GluSNF4 and RCamp3 both with two photon. Expression is rather dense, causing a little background signal.

- Experiment started at about 15:42

Maedeh circuled the soma, a few proximal dendrites and an apical dendrites. So this experiment should yield calcium (red) + synaptic inputs (green).

We had a false start of about a min. The stimulus started but SLAP2 somehow stopped recording after 30 s. We reinitialized the system and restarted. Then bonsai had an issue with cameras so we immediately restarted the experiment again. On the third trial, we had some motion artifacts initially. Maedeh fine tuned the online motion correction algorithm initially. Mice was moving its mouth very quickly at the onset of the experiment probably causing those issues. The experiment stabilitized a few minutes in. PLane with soma tend to have a little more motion artifacts from time to time. Apical plane is more stable overall.

MAJOR ISSUE: Slap2 stopped acquiring data at about 4:18PM. Maedeh pushed it forward manually at 4:22PM So this experiment has a whole in the middle. Bonsai was continuing recording.

- Apical dendrites is imaged in DMD1, Soma in DMD2, depth 155 um for soma. 
- Experiment went smoothly. We ended on time at 4:34

We experimented with the online motion correction after the session ended. We understood that the issue was the presence of the soma in the set of ROI confusing the motion correction algorithm. Turning it off greatly improve the online correction. We will have to follow that procedure later on : Turn off motion correction on somatic ROI and focus on dendrites to estimate X,Y,Z motion.

Checking the eye tracking data seems to show some kind of floater inside the eye. It is not entirely clear what this is. Could be a tiny cataract.

5:20 Stacks was acquired. We had to reestart it as the software crashed mid-way.