Fast, multiplane line-scan confocal microscopy using axially distributed slits

The inherent constraints on resolution, speed and field of view have hindered the development of high-speed, three-dimensional microscopy techniques over large scales. Here, we present a multiplane line-scan imaging strategy, which uses a series of axially distributed reflecting slits to probe different depths within a sample volume. Our technique enables the simultaneous imaging of an optically sectioned image stack with a single camera at frame rates of hundreds of hertz, without the need for axial scanning. We demonstrate the applicability of our system to monitor fast dynamics in biological samples by performing calcium imaging of neuronal activity in mouse brains and voltage imaging of cardiomyocytes in cardiac samples. (c) 2021 Optical Society of America under the terms of the OSA Open Access Publishing Agreement

Automated summary

This study presents a multiplane line-scan imaging strategy using reflecting slits to probe different depths, enabling simultaneous imaging of optically sectioned image stack with a single camera at frame rates of hundreds of hertz. The system is demonstrated to be applicable for monitoring fast dynamics in biological samples.