Two-photon line excitation array detection (2p-LEAD) microscopy for monitoring in vivo neural activity

Capturing the fastest biological events in the mammalian brain, such as hemodynamic flow and neuronal signal propagation, requires high temporal resolution and imaging through scattering biological tissue. To fully resolve these functional dynamics, we present an improved version of 2-photon Line Excitation Array Detection (2p-LEAD) microscopy. Through shaping the 1035 nm excitation beam into a line, 2p-LEAD can operate at 4000 FPS, using a linear array of PMTs to collect the resulting fluorescence. With the replacement of the scanning mirror for an acousto-optic deflector, 2p-LEAD holds the potential of achieving kilohertz volumetric imaging over the visual cortex of the mice brain.