title: Enhancing image quality in cleared tissue with adaptive optics.
abstract: Our ability to see fine detail at depth in tissues is limited by scattering and other refractive characteristics of the tissue. For fixed tissue, we can limit scattering with a variety of clearing protocols. This allows us to see deeper but not necessarily clearer. Refractive aberrations caused by the bulk index of refraction of the tissue and its variations continue to limit our ability to see fine detail. Refractive aberrations are made up of spherical and other Zernike modes, which can be significant at depth. Spherical aberration that is common across the imaging field can be corrected using an objective correcting collar, although this can require manual intervention. Other aberrations may vary across the imaging field and can only be effectively corrected using adaptive optics. Adaptive optics can also correct other aberrations simultaneously with the spherical aberration, eliminating manual intervention and speeding imaging. We use an adaptive optics two-photon microscope to examine the impact of the spherical and higher order aberrations on imaging and contrast the effect of compensating only for spherical aberration against compensating for the first 22 Zernike aberrations in two tissue types. Increase in image intensity by 1.6× and reduction of root mean square error by 3× are demonstrated.
authors.affiliation: University of California Santa Cruz, W.M. Keck Center for Adaptive Optical Microscopy, Baskin Engineering, 1154 High Street, Santa Cruz, California 95064, United States.
authors.forename: Marc R
authors.initials: MR
authors.lastname: Reinig
authors.affiliation: University of California Santa Cruz, W.M. Keck Center for Adaptive Optical Microscopy, Baskin Engineering, 1154 High Street, Santa Cruz, California 95064, United States.
authors.forename: Samuel W
authors.initials: SW
authors.lastname: Novak
authors.affiliation: University of California Santa Cruz, W.M. Keck Center for Adaptive Optical Microscopy, Baskin Engineering, 1154 High Street, Santa Cruz, California 95064, United States.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Tao
authors.affiliation: University of California, California Nanosystems Institute, Advanced Light Microscopy/Spectroscopy Laboratory, 570 Westwood Plaza, Building 114, Los Angeles, California 90095, United States.
authors.forename: Laurent A
authors.initials: LA
authors.lastname: Bentolila
authors.affiliation: UCLA Brain Mapping Center, 660 Charles E. Young Drive South, Los Angeles, California 90095, United States.
authors.forename: Dustin G
authors.initials: DG
authors.lastname: Roberts
authors.affiliation: UCLA Neurology, 710 Westwood Plaza, PO Box 951769, 4256 Los Angeles, California 90095-1769, United States.
authors.forename: Allan
authors.initials: A
authors.lastname: MacKenzie-Graham
authors.affiliation: University of California, Neuroscience Research Institute, Microscopy Facility, 3087 Calle Rosales, Santa Barbara, California 93105, United States.
authors.forename: Sirie E
authors.initials: SE
authors.lastname: Godshalk
authors.affiliation: UCSB's Office of Technology and Industry Alliances, 342 Lagoon Road, Mail Code 2055, Santa Barbara, California 93106-2055, United States.
authors.forename: Mary A
authors.initials: MA
authors.lastname: Raven
authors.affiliation: Life Sciences Division, Berkeley Drosophila Transcription Network Project, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, California 94720, United States.
authors.forename: David W
authors.initials: DW
authors.lastname: Knowles
authors.affiliation: University of California Santa Cruz, W.M. Keck Center for Adaptive Optical Microscopy, Baskin Engineering, 1154 High Street, Santa Cruz, California 95064, United States.
authors.forename: Joel
authors.initials: J
authors.lastname: Kubby
mesh_terms: D000818:Animals; D001921:Brain / Q000000981:diagnostic imaging; D004867:Equipment Design; D007089:Image Enhancement / Q000379:methods*; D008164:Luminescent Proteins; D051379:Mice; D008822:Mice, Transgenic; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D016501:Neurites / Q000737:chemistry / Q000378:metabolism; D013116:Spinal Cord / Q000000981:diagnostic imaging
chemical_list: D008164:Luminescent Proteins