The Cousa objective: a long working distance air objective for multiphoton imaging in vivo

Kavli Affiliate: Michael Higley, Alipasha Vaziri, Kristina Nielsen

| Authors: Che-Hang Yu, Yiyi Yu, Liam M. Adsit, Jeremy T. Chang, Jad Barchini, Andrew H. Moberly, Hadas Benisty, Jinkyung Kim, Brent K Young, Kathleen Heng, Deano M. Farinella, Austin Leikvoll, Rishaab Pavan, Rachel Vistein, Brandon R. Nanfito, David G. C. Hildebrand, Santiago Otero-Coronel, Alipasha Vaziri, Jeffery L. Goldberg, Anthony J. Ricci, David Fitzpatrick, Jessica A Cardin, Michael Higley, Gordon B. Smith, Prakash Kara, Kristina J. Nielsen, Ikuko T. Smith and Spencer LaVere Smith

| Summary:

Multiphoton microscopy can resolve fluorescent structures and dynamics deep in scattering tissue, but applying this technique in vivo can be limited by short working distance water-immersion objectives. 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 > 4 mm^2 field-of-view with submicron lateral resolution, and is compatible with commonly used multiphoton imaging systems. We share the full optical prescription, and report performance including in vivo 2-photon and 3-photon imaging in a range of species and preparations.

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