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The Cousa objective: a long-working distance air objective for multiphoton imaging in vivo.
Yu, Che-Hang; Yu, Yiyi; Adsit, Liam M; Chang, Jeremy T; Barchini, Jad; Moberly, Andrew H; Benisty, Hadas; Kim, Jinkyung; Young, Brent K; Heng, Kathleen; Farinella, Deano M; Leikvoll, Austin; Pavan, Rishaab; Vistein, Rachel; Nanfito, Brandon R; Hildebrand, David G C; Otero-Coronel, Santiago; Vaziri, Alipasha; Goldberg, Jeffrey L; Ricci, Anthony J; Fitzpatrick, David; Cardin, Jessica A; Higley, Michael J; Smith, Gordon B; Kara, Prakash; Nielsen, Kristina J; Smith, Ikuko T; Smith, Spencer LaVere.
Affiliation
  • Yu CH; Department of Electrical and Computer Engineering, University of California Santa Barbara, Santa Barbara, CA, USA. chehangyu@ucsb.edu.
  • Yu Y; Department of Electrical and Computer Engineering, University of California Santa Barbara, Santa Barbara, CA, USA.
  • Adsit LM; Department of Molecular, Cellular, and Developmental Biology, University of California Santa Barbara, Santa Barbara, CA, USA.
  • Chang JT; Max Planck Florida Institute for Neuroscience, Jupiter, FL, USA.
  • Barchini J; Max Planck Florida Institute for Neuroscience, Jupiter, FL, USA.
  • Moberly AH; Department of Neuroscience, Yale University, New Haven, CT, USA.
  • Benisty H; Department of Neuroscience, Yale University, New Haven, CT, USA.
  • Kim J; Department of Otolaryngology, Washington University School of Medicine, St. Louis, MO, USA.
  • Young BK; Spencer Center for Vision Research, Byers Eye Institute, School of Medicine, Stanford University, Palo Alto, CA, USA.
  • Heng K; Spencer Center for Vision Research, Byers Eye Institute, School of Medicine, Stanford University, Palo Alto, CA, USA.
  • Farinella DM; Neurosciences Interdepartmental Program, Stanford University, Stanford, CA, USA.
  • Leikvoll A; Department of Neuroscience, University of Minnesota, Minneapolis, MN, USA.
  • Pavan R; Department of Neuroscience, University of Minnesota, Minneapolis, MN, USA.
  • Vistein R; Department of Biomedical Engineering, University of Minnesota, Minneapolis, MN, USA.
  • Nanfito BR; Department of Molecular and Comparative Pathobiology, and Zanvyl Krieger Mind/Brain Institute, Johns Hopkins University, Baltimore, MD, USA.
  • Hildebrand DGC; Solomon H. Snyder Department of Neuroscience, and Zanvyl Krieger Mind/Brain Institute, Johns Hopkins University, Baltimore, MD, USA.
  • Otero-Coronel S; Laboratory of Neural Systems, The Rockefeller University, New York, NY, USA.
  • Vaziri A; Laboratory of Neural Systems, The Rockefeller University, New York, NY, USA.
  • Goldberg JL; Laboratory of Neurotechnology and Biophysics, The Rockefeller University, New York, NY, USA.
  • Ricci AJ; Kavli Neural Systems Institute, The Rockefeller University, New York, NY, USA.
  • Fitzpatrick D; Laboratory of Neurotechnology and Biophysics, The Rockefeller University, New York, NY, USA.
  • Cardin JA; Kavli Neural Systems Institute, The Rockefeller University, New York, NY, USA.
  • Higley MJ; Spencer Center for Vision Research, Byers Eye Institute, School of Medicine, Stanford University, Palo Alto, CA, USA.
  • Smith GB; Department of Otolaryngology, Stanford University School of Medicine, Stanford University, Stanford, CA, USA.
  • Kara P; Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford University, Stanford, CA, USA.
  • Nielsen KJ; Max Planck Florida Institute for Neuroscience, Jupiter, FL, USA.
  • Smith IT; Department of Neuroscience, Yale University, New Haven, CT, USA.
  • Smith SL; Department of Neuroscience, Yale University, New Haven, CT, USA.
Nat Methods ; 21(1): 132-141, 2024 Jan.
Article in En | MEDLINE | ID: mdl-38129618
ABSTRACT
Multiphoton microscopy can resolve fluorescent structures and dynamics deep in scattering tissue and has transformed neural imaging, but applying this technique in vivo can be limited by the mechanical and optical constraints of conventional objectives. Short working distance objectives can collide with compact surgical windows or other instrumentation and preclude imaging. Here we present an ultra-long working distance (20 mm) air objective called the Cousa objective. It is optimized for performance across multiphoton imaging wavelengths, offers a more than 4 mm2 field of view with submicrometer lateral resolution and is compatible with commonly used multiphoton imaging systems. A novel mechanical design, wider than typical microscope objectives, enabled this combination of specifications. We share the full optical prescription, and report performance including in vivo two-photon and three-photon imaging in an array of species and preparations, including nonhuman primates. The Cousa objective can enable a range of experiments in neuroscience and beyond.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Microscopy, Fluorescence, Multiphoton / Coloring Agents Limits: Animals Language: En Journal: Nat Methods / Nat. methods / Nature methods Journal subject: TECNICAS E PROCEDIMENTOS DE LABORATORIO Year: 2024 Document type: Article Affiliation country: Estados Unidos Country of publication: Estados Unidos
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Microscopy, Fluorescence, Multiphoton / Coloring Agents Limits: Animals Language: En Journal: Nat Methods / Nat. methods / Nature methods Journal subject: TECNICAS E PROCEDIMENTOS DE LABORATORIO Year: 2024 Document type: Article Affiliation country: Estados Unidos Country of publication: Estados Unidos