Relationship between simultaneously recorded spiking activity and fluorescence signal in GCaMP6 transgenic mice

Fluorescent calcium indicators are often used to investigate neural dynamics, but the relationship between fluorescence and action potentials (APs) remains unclear. Most APs can be detected when the soma almost fills the microscope's field of view, but calcium indicators are used to image populations of neurons, necessitating a large field of view, generating fewer photons per neuron, and compromising AP detection. Here, we characterized the AP-fluorescence transfer function in vivo for 48 layer 2/3 pyramidal neurons in primary visual cortex, with simultaneous calcium imaging and cell-attached recordings from transgenic mice expressing GCaMP6s or GCaMP6f. While most APs were detected under optimal conditions, under conditions typical of population imaging studies, only a minority of 1 AP and 2 AP events were detected (often <10% and similar to 20-30%, respectively), emphasizing the limits of AP detection under more realistic imaging conditions.

Automated summary

Fluorescent calcium indicators are commonly used to study neural dynamics, but their relationship with action potentials remains unclear. While most APs can be detected under optimal conditions, limitations arise when imaging populations of neurons, raising challenges for accurate AP detection.