Multibeam two-photon microscopy for multispectral ultrahigh-throughput imaging

A multiplane two-photon microscope with non-descanned detection provides imaging speeds up to 200 MP/s with four spectral channels (800 MSpectra/s) with area imaging speeds of up to 30 mm2/s. This enables high volumetric throughput deep-tissue imaging at speeds approaching light-sheet imaging without sacrificing resilience to scattering and high axial resolution. Multiplane excitation is achieved by multifocal scanner-synchronous strip-scanning with tilted translation, while spherical lens arrays with custom SiPM array boards provide non-descanned detection that is both inexpensive and easy to spectrally multiplex.