title: SPED Light Sheet Microscopy: Fast Mapping of Biological System Structure and Function.
abstract: The goal of understanding living nervous systems has driven interest in high-speed and large field-of-view volumetric imaging at cellular resolution. Light sheet microscopy approaches have emerged for cellular-resolution functional brain imaging in small organisms such as larval zebrafish, but remain fundamentally limited in speed. Here, we have developed SPED light sheet microscopy, which combines large volumetric field-of-view via an extended depth of field with the optical sectioning of light sheet microscopy, thereby eliminating the need to physically scan detection objectives for volumetric imaging. SPED enables scanning of thousands of volumes-per-second, limited only by camera acquisition rate, through the harnessing of optical mechanisms that normally result in unwanted spherical aberrations. We demonstrate capabilities of SPED microscopy by performing fast sub-cellular resolution imaging of CLARITY mouse brains and cellular-resolution volumetric Ca(2+) imaging of entire zebrafish nervous systems. Together, SPED light sheet methods enable high-speed cellular-resolution volumetric mapping of biological system structure and function.
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; CNC Program, Stanford University, Stanford, CA 94305, USA.
authors.forename: Raju
authors.initials: R
authors.lastname: Tomer
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; CNC Program, Stanford University, Stanford, CA 94305, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Lovett-Barron
authors.affiliation: CNC Program, Stanford University, Stanford, CA 94305, USA; Department of Electrical Engineering, Stanford University, Stanford, CA 94305, USA.
authors.forename: Isaac
authors.initials: I
authors.lastname: Kauvar
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; CNC Program, Stanford University, Stanford, CA 94305, USA.
authors.forename: Aaron
authors.initials: A
authors.lastname: Andalman
authors.affiliation: CNC Program, Stanford University, Stanford, CA 94305, USA; Department of Chemical and Systems Biology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Vanessa M
authors.initials: VM
authors.lastname: Burns
authors.affiliation: CNC Program, Stanford University, Stanford, CA 94305, USA.
authors.forename: Sethuraman
authors.initials: S
authors.lastname: Sankaran
authors.affiliation: CNC Program, Stanford University, Stanford, CA 94305, USA.
authors.forename: Logan
authors.initials: L
authors.lastname: Grosenick
authors.affiliation: Department of Computer Science, Stanford University, Stanford, CA 94305, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Broxton
authors.affiliation: CNC Program, Stanford University, Stanford, CA 94305, USA; Department of Electrical Engineering, Stanford University, Stanford, CA 94305, USA.
authors.forename: Samuel
authors.initials: S
authors.lastname: Yang
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; CNC Program, Stanford University, Stanford, CA 94305, USA; Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA; Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA 94305, USA. Electronic address: deissero@stanford.edu.
authors.forename: Karl
authors.initials: K
authors.lastname: Deisseroth
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology; D007091:Image Processing, Computer-Assisted / Q000379:methods; D007814:Larva / Q000166:cytology; D051379:Mice; D008853:Microscopy / Q000379:methods*; D009420:Nervous System / Q000166:cytology*; D016501:Neurites / Q000648:ultrastructure; D015027:Zebrafish / Q000254:growth & development