Swept confocally-aligned planar excitation (SCAPE) microscopy for high-speed volumetric imaging of behaving organisms

Abstract

We report a new 3D microscopy technique that allows volumetric imaging of living samples at ultra-high speeds: Swept, confocally-aligned planar excitation (SCAPE) microscopy. While confocal and two-photon microscopy have revolutionized biomedical research, current implementations are costly, complex and limited in their ability to image 3D volumes at high speeds. Light-sheet microscopy techniques using two-objective, orthogonal illumination and detection require a highly constrained sample geometry, and either physical sample translation or complex synchronization of illumination and detection planes. In contrast, SCAPE microscopy acquires images using an angled, swept light-sheet in a single-objective, en-face geometry. Unique confocal descanning and image rotation optics map this moving plane onto a stationary high-speed camera, permitting completely translationless 3D imaging of intact samples at rates exceeding 20 volumes per second. We demonstrate SCAPE microscopy by imaging spontaneous neuronal firing in the intact brain of awake behaving mice, as well as freely moving transgenic Drosophila larvae.

The use of computational modeling to link sensory processing with behavior in Drosophila

“…We expect that methodological innovations -both experimental and theoretical -will further increase the utility of computational modeling in this model system. Imaging technology now permits recording from larger numbers of Drosophila neurons simultaneously (Bouchard et al 2015;Harris et al 2015;Lu et al 2016;Aimon et al 2016) or from subsets of neurons in freely behaving flies (Grover et al 2016). Computational models applied to such data will facilitate both interpreting population neural activity and connecting neural activity with behavior.…”

The real-time technicolour living brain

“…But by turning the sheet on an angle and using a single mirror to sweep it across the volume of interest, Hillman's group achieved a rate of 20 times per second. Hillman calls the technique swept confocally aligned planar excitation, or SCAPE, and her team has used it to visualize dozens of distinct firing patterns in the brains of awake mice 8 . The technology has been licensed to Leica Microsystems in Wetzlar, Germany.…”

In vivo volumetric imaging of biological dynamics in deep tissue via wavefront engineering

“…The parallel detection scheme of these methods benefits high-speed imaging. However, it only works well for transparent biological systems [7], such as cultured cells, C Elegans and zebrafish larvae.…”