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

Yu, C.-H.; Yu, Y.; Adsit, L. M.; Chang, J. T.; Barchini, J.; Moberly, A. H.; Benisty, H.; Kim, J.; Ricci, A. J.; Fitzpatrick, D.; Cardin, J. A.; Higley, M.; Smith, G. B.; Nielsen, K. J.; Smith, I. T.; Smith, S. L.

doi:10.1101/2022.11.06.515343

bioRxiv · 10.1101/2022.11.06.515343

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

Yu, C.-H.·Yu, Y.·Adsit, L. M.·Chang, J. T.·Barchini, J.·Moberly, A. H.·Benisty, H.·Kim, J.·Ricci, A. J.·Fitzpatrick, D.·Cardin, J. A.·Higley, M.·Smith, G. B.·Nielsen, K. J.·Smith, I. T.·Smith, S. L.

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Abstract

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 mm2 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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Yu, C.-H., Yu, Y., Adsit, L. M., Chang, J. T., Barchini, J., Moberly, A. H., Benisty, H., Kim, J., Ricci, A. J., Fitzpatrick, D., Cardin, J. A., Higley, M., Smith, G. B., Nielsen, K. J., Smith, I. T., Smith, S. L.. 2022-11-06. The Cousa objective: a long working distance air objective for multiphoton imaging in vivo. https://doi.org/10.1101/2022.11.06.515343

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