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Reflective multi-immersion microscope objectives inspired by the Schmidt telescope

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2024

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Voigt, Fabian F.
Reuss, Anna Maria
Naert, Thomas
Hildebrand, Sven
Schaettin, Martina
Hotz, Adriana L.
Whitehead, Lachlan
Neuhauss, Stephan C. F.
Roebroeck, Alard
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Nature Biotechnology. Springer. 2024, 42(1), pp. 65-71. ISSN 1087-0156. eISSN 1546-1696. Available under: doi: 10.1038/s41587-023-01717-8

Zusammenfassung

Imaging large, cleared samples requires microscope objectives that combine a large field of view (FOV) with a long working distance (WD) and a high numerical aperture (NA). Ideally, such objectives should be compatible with a wide range of immersion media, which is challenging to achieve with conventional lens-based objective designs. Here we introduce the multi-immersion ‘Schmidt objective’ consisting of a spherical mirror and an aspherical correction plate as a solution to this problem. We demonstrate that a multi-photon variant of the Schmidt objective is compatible with all homogeneous immersion media and achieves an NA of 1.08 at a refractive index of 1.56, 1.1-mm FOV and 11-mm WD. We highlight its versatility by imaging cleared samples in various media ranging from air and water to benzyl alcohol/benzyl benzoate, dibenzyl ether and ethyl cinnamate and by imaging of neuronal activity in larval zebrafish in vivo. In principle, the concept can be extended to any imaging modality, including wide-field, confocal and light-sheet microscopy.

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570 Biowissenschaften, Biologie

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ISO 690VOIGT, Fabian F., Anna Maria REUSS, Thomas NAERT, Sven HILDEBRAND, Martina SCHAETTIN, Adriana L. HOTZ, Lachlan WHITEHEAD, Armin BAHL, Stephan C. F. NEUHAUSS, Alard ROEBROECK, 2024. Reflective multi-immersion microscope objectives inspired by the Schmidt telescope. In: Nature Biotechnology. Springer. 2024, 42(1), pp. 65-71. ISSN 1087-0156. eISSN 1546-1696. Available under: doi: 10.1038/s41587-023-01717-8
BibTex
@article{Voigt2024Refle-67638,
  year={2024},
  doi={10.1038/s41587-023-01717-8},
  title={Reflective multi-immersion microscope objectives inspired by the Schmidt telescope},
  number={1},
  volume={42},
  issn={1087-0156},
  journal={Nature Biotechnology},
  pages={65--71},
  author={Voigt, Fabian F. and Reuss, Anna Maria and Naert, Thomas and Hildebrand, Sven and Schaettin, Martina and Hotz, Adriana L. and Whitehead, Lachlan and Bahl, Armin and Neuhauss, Stephan C. F. and Roebroeck, Alard}
}
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mesoSPIM datasets were acquired using mesospim-control 1.7.2, a custom open-source image acquisition software. Code for PSF analysis is available upon reasonable request.
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