Swept confocally aligned planar excitation (SCAPE) microscopy is a method for high-speed, cellular level imaging of living, intact tissues. SCAPE combines unique features of both light sheet microscopy and confocal scanning to image at over 20 volumes per second in intact, unmounted samples, requiring no movement of the objective lens or sample, providing low photobleaching and high signal to noise, and having relatively simple and inexpensive instrumentation. Under an ongoing BRAIN initiative U01 we have made dramatic improvements to SCAPE imaging performance, while fostering and supporting the adoption of SCAPE by a wide range of neuroscience collaborators working in diverse model systems. Through application-driven optimization, SCAPE has already contributed to several fundamental neuroscience discoveries. Our work has also demonstrated the feasibility of two-photon SCAPE, bringing the potential to image at very high volumetric imaging speeds deeper into the intact mammalian brain. Here we propose to continue this highly productive trajectory, expanding our research in three important directions 1) To continue innovation by implementing multiple innovative second-generation variants of SCAPE to deliver closed-loop imaging for small organism behavior and optogenetics, ultrahigh resolution imaging for C. elegans, meso-scale imaging for large-area, depth-resolved imaging in mammalian brain, near infrared implementations to leverage new red and NIR-shifted indications, and continued development of two different two-photon SCAPE implementations. 2) Continued and expanded development and support of new applications in close collaboration with leading neuroscientists including crawling larval Drosophila and adult behaving Drosophila brain, C. elegans, zebrafish brain, mouse olfactory epithelium, awake behaving mouse and marmoset brain. 3) Expanding dissemination by supporting an initial cohort of researchers to develop their own `Open-SCAPE' systems ? a version of SCAPE that we are optimizing for simple, well-documented assemble, maintenance and use. Establishing and supporting this first international community of SCAPE users will be an important step in optimizing our Open-SCAPE design. Recipients will also quickly be able to integrate SCAPE into their research, while generating a new knowledge base of SCAPE expertise, experimental paradigms and analysis methods that can be more broadly shared as the community grows. Combined, these efforts will enable expanded access to robust SCAPE platforms for neuroscience research, in parallel with ongoing next-generation innovation, optimizations and diversification of applications delivering vital new high-speed cellular imaging capabilities across the spectrum of neuroscience research.
This project aims to continue and extend development of swept confocally aligned planar excitation (SCAPE) microscopy, a technique that can image living samples in 3D at very high speeds. We will both develop next- generation versions of SCAPE and support collaborators to optimize SCAPE for their neuroscience experiments in addition to helping others to build their own SCAPE systems.