Super-resolution upgrade for deep tissue imaging featuring simple implementation
Abstract Deep tissue imaging with high contrast close to or even below the optical resolution limit is still challenging due to optical aberrations and scattering introduced by dense biological samples. This results in high complexity and cost of microscopes that can facilitate such challenges. Here...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-06-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-60744-y |
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| _version_ | 1849432594421121024 |
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| author | Patrick Byers Thomas Kellerer Miaomiao Li Zhifen Chen Thomas Huser Thomas Hellerer |
| author_facet | Patrick Byers Thomas Kellerer Miaomiao Li Zhifen Chen Thomas Huser Thomas Hellerer |
| author_sort | Patrick Byers |
| collection | DOAJ |
| description | Abstract Deep tissue imaging with high contrast close to or even below the optical resolution limit is still challenging due to optical aberrations and scattering introduced by dense biological samples. This results in high complexity and cost of microscopes that can facilitate such challenges. Here, we demonstrate a cost-effective and simple to implement method to turn most two-photon laser-scanning microscopes into a super-resolution microscope for deep tissue imaging. We realize this by adding inexpensive optical devices, namely a cylindrical lens, a field rotator, and a sCMOS camera to these systems. By combining two-photon excitation with patterned line-scanning and subsequent image reconstruction, we achieve imaging of sub-cellular structures in Pinus radiata, mouse heart muscle and zebrafish. In addition, the penetration depth of super-resolved imaging in highly scattering tissue is considerably extended by using the camera’s lightsheet shutter mode. The flexibility of our method allows the examination of a variety of thick samples with a variety of fluorescent markers and microscope objective lenses. Thus, with a cost-efficient modification of a multi-photon microscope, an up to twofold resolution enhancement is demonstrated down to at least 70μm deep in tissue. |
| format | Article |
| id | doaj-art-db80ad288eaa4d40a83ef42df134143b |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-db80ad288eaa4d40a83ef42df134143b2025-08-20T03:27:18ZengNature PortfolioNature Communications2041-17232025-06-0116111210.1038/s41467-025-60744-ySuper-resolution upgrade for deep tissue imaging featuring simple implementationPatrick Byers0Thomas Kellerer1Miaomiao Li2Zhifen Chen3Thomas Huser4Thomas Hellerer5Multiphoton Imaging Lab, Munich University of Applied SciencesMultiphoton Imaging Lab, Munich University of Applied SciencesDepartment of Cardiology, German Heart Center, TUM University Hospital, TUM School of Medicine and Health, Technical University MunichDepartment of Cardiology, German Heart Center, TUM University Hospital, TUM School of Medicine and Health, Technical University MunichBiomolecular Photonics, Department of Physics, Bielefeld UniversityMultiphoton Imaging Lab, Munich University of Applied SciencesAbstract Deep tissue imaging with high contrast close to or even below the optical resolution limit is still challenging due to optical aberrations and scattering introduced by dense biological samples. This results in high complexity and cost of microscopes that can facilitate such challenges. Here, we demonstrate a cost-effective and simple to implement method to turn most two-photon laser-scanning microscopes into a super-resolution microscope for deep tissue imaging. We realize this by adding inexpensive optical devices, namely a cylindrical lens, a field rotator, and a sCMOS camera to these systems. By combining two-photon excitation with patterned line-scanning and subsequent image reconstruction, we achieve imaging of sub-cellular structures in Pinus radiata, mouse heart muscle and zebrafish. In addition, the penetration depth of super-resolved imaging in highly scattering tissue is considerably extended by using the camera’s lightsheet shutter mode. The flexibility of our method allows the examination of a variety of thick samples with a variety of fluorescent markers and microscope objective lenses. Thus, with a cost-efficient modification of a multi-photon microscope, an up to twofold resolution enhancement is demonstrated down to at least 70μm deep in tissue.https://doi.org/10.1038/s41467-025-60744-y |
| spellingShingle | Patrick Byers Thomas Kellerer Miaomiao Li Zhifen Chen Thomas Huser Thomas Hellerer Super-resolution upgrade for deep tissue imaging featuring simple implementation Nature Communications |
| title | Super-resolution upgrade for deep tissue imaging featuring simple implementation |
| title_full | Super-resolution upgrade for deep tissue imaging featuring simple implementation |
| title_fullStr | Super-resolution upgrade for deep tissue imaging featuring simple implementation |
| title_full_unstemmed | Super-resolution upgrade for deep tissue imaging featuring simple implementation |
| title_short | Super-resolution upgrade for deep tissue imaging featuring simple implementation |
| title_sort | super resolution upgrade for deep tissue imaging featuring simple implementation |
| url | https://doi.org/10.1038/s41467-025-60744-y |
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