PUBLICATION

Reflective multi-immersion microscope objectives inspired by the Schmidt telescope

Authors
Voigt, F.F., Reuss, A.M., Naert, T., Hildebrand, S., Schaettin, M., Hotz, A.L., Whitehead, L., Bahl, A., Neuhauss, S.C.F., Roebroeck, A., Stoeckli, E.T., Lienkamp, S.S., Aguzzi, A., Helmchen, F.
ID
ZDB-PUB-230331-51
Date
2023
Source
Nature Biotechnology   42(1): 65-71 (Journal)
Registered Authors
Hotz, Adriana, Neuhauss, Stephan
Keywords
none
MeSH Terms
  • Animals
  • Immersion
  • Microscopy/methods
  • Telescopes*
  • Zebrafish
PubMed
36997681 Full text @ Nat Biotechnol.
Abstract
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.
Genes / Markers
Figures
Show all Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping