High-speed three-dimensional random access scanning with a linear SLM

Abstract

High-speed volumetric imaging is crucial for observing fast and distributed processes such as neuronal activity. Multiphoton microscopy helps to mitigate scattering effects inside tissue, but the standard raster scanning approach limits achievable volume rates. Random access point scanning can lead to a considerable speed-up by sampling only pre-selected locations, but existing techniques based on acousto-optic deflectors are still limited to a point rate of up to similar to 50 kHz. This limits the number of parallel targets at the high acquisition rates necessary, for example, in voltage imaging or imaging of fast synaptic events. Here, we introduce SPARCLS, a method for 3D random access point scanning at up to 340 kHz using a single 1D phase modulator. We show the potential of this method by imaging synaptic events with fluorescent glutamate sensors in mammalian organotypic slices as well as in zebrafish larvae.