title: 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 transgenicDrosophilalarvae.
authors: Laboratory for Functional Optical Imaging, Departments of Biomedical Engineering and Radiology, Columbia University, New York, NY 10027.
authors: Matthew B
authors: MB
authors: Bouchard
authors: Laboratory for Functional Optical Imaging, Departments of Biomedical Engineering and Radiology, Columbia University, New York, NY 10027.
authors: Venkatakaushik
authors: V
authors: Voleti
authors: Mann Lab, Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032.
authors: César S
authors: CS
authors: Mendes
authors: Bruno Lab, Department of Neuroscience, Columbia University, New York, NY 10032.
authors: Clay
authors: C
authors: Lacefield
authors: Department of Physiology and Cellular Biophysics, Department of Neuroscience, College of Physicians and Surgeons, Columbia University Medical Center, New York, NY 10032.
authors: Wesley B
authors: WB
authors: Grueber
authors: Mann Lab, Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032.
authors: Richard S
authors: RS
authors: Mann
authors: Bruno Lab, Department of Neuroscience, Columbia University, New York, NY 10032.
authors: Randy M
authors: RM
authors: Bruno
authors: Laboratory for Functional Optical Imaging, Departments of Biomedical Engineering and Radiology, Columbia University, New York, NY 10027.
authors: Elizabeth M C
authors: EM
authors: Hillman