Widefield and two-photon calcium imaging

We make a craniotomy so it begins about 0.25-0.5 mm anterior to lambda (the craniotomy is 3 mm diameter). This should capture A1/AAF, which begin about 1.5 mm anterior to lambda. The craniotomy is also centered on the temporal ridge (medial-lateral axis).

There are no stereotactic coordinates for primary auditory cortex. This location needs to be determined physiologically, which we do here by playing tones of different frequencies/intensities during widefield epifluorescence calcium imaging and calculating the frequency tuning across the space of our craniotomy.

For further details, please see:

• Romero, et al. Cerebral Cortex 2020. https://doi.org/10.1093/cercor/bhz190
• Narayanan, et al. Cerebral Cortex 2023. https://doi.org/10.1093/cercor/bhac275