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. 2023 Dec 21;21(1):132–141. doi: 10.1038/s41592-023-02098-1

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a, In behaving marmosets, the Cousa objective facilitates imaging by replacing water immersion with air and providing ample open space around the cranial window. b, In ferrets, it can be challenging to access the neurons of interest. A short WD objective (Nikon ×16/0.8 NA, 3 mm WD in water) cannot even access the neurons of interest due to the short WD and collisions with the walls of the cranial window. The Cousa provides a large FOV and activity measurement, another long WD air objective provides only a smaller FOV (at a different angle) and weaker signals. Imaging depth is roughly 300 µm. c, In ferrets, with another type of imaging window, the Cousa provides a large FOV for measuring orientation tuning across orientation columns. d, Similarly, in tree shrew, the Cousa provides a large FOV for resolving individual neurons across orientation domains. e, In a challenging preparation, the Cousa enabled two-photon imaging through the lens and entire eye to the retina of an intact porcine eye, including retinal ganglion cell bodies (arrows) and single axon fibers (triangles).