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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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Extended Data Fig. 2 — A schematic shows the ray-traces in the collection light path for the Cousa objective. Fluorescence photons with wavelengths in the visible band (450 nm, 550 nm, and 650 nm) are emitted from the imaging plane within the 2 mm nominal FOV (zoom in view on the left), passing through the Cousa objective. These photons are further collected and guided by the optical relay, and reach at the cathode of the photomultiplier (PMT). The optical relay is constructed simply with two off-the-shelf singlet lenses from OptoSigma (011-2570-a55) and Thorlabs (LA1805-A). The zoom in view in the bottom-right shows that all of the emitted photons (rays) in the visible spectrum reach at the detector surface within a 3 mm wide spread.