Adapting upright light sheet fluorescence microscopy for imaging at air-liquid interface
Abstract Light sheet fluorescence microscopy (LSFM) is increasingly appreciated as the gold standard for gentle, volumetric imaging with fast acquisition speeds and/or long imaging durations. However, the often-constrained sample space of these microscopes has precluded a specific class of biological specimens from being studied with these tools: those requiring an air-liquid interface (ALI). Here, we present a device for robust imaging at ALI on an upright light sheet microscope with dipping objectives. We demonstrate the system using three relevant use-cases: ex vivo embryonic mouse salivary glands, human epidermal equivalent cultures, and in vivo adult Drosophila melanogaster brains. While the device presented is engineered for one specific light sheet microscope design, it provides a blueprint for easy adaptation to other systems. In doing so, it can potentially spur the use of LSFM for model systems that have so far been unable to take advantage of this powerful technology.
Authors
- T. Tsuchiya (ORCID: https://orcid.org/0000-0003-0096-4275)
- Sara N. Stahley (ORCID: https://orcid.org/0000-0002-2316-8354)
- Owen Puls (ORCID: https://orcid.org/0000-0002-5097-3420)
- Andrew P. Kowalczyk (ORCID: https://orcid.org/0000-0001-6438-7863)
- Navaneetha Krishnan Bharathan (ORCID: https://orcid.org/0000-0001-8340-5561)
- Shaohe Wang (ORCID: https://orcid.org/0000-0001-9522-2359)
- Micaela Rodríguez-Caron (ORCID: https://orcid.org/0009-0007-2011-7002)
- Francisco Tassara
- Shunze Yao
- William Giang
- Juan I. Ispizua (ORCID: https://orcid.org/0000-0002-4183-5392)
- Chad M. Hobson (ORCID: https://orcid.org/0000-0003-3033-5728)
- Jesse S. Aaron (ORCID: https://orcid.org/0000-0002-7543-9590)
- E. Angelo Morales
- Rachel M. Lee
- Teng-Leong Chew
- Kenji Izumi
- Ellen Quarles
- M. Fernanda Ceriani
Institutions
- Pennsylvania State University (US)
- Tokai University (JP)
- Howard Hughes Medical Institute (US)
- Janelia Research Campus (US)
- University of Washington (US)
- Peking University (CN)
- Fundación Instituto Leloir (AR)
Publication Details
- Journal
- npj Imaging
- Published
- 2026-07-22
- DOI
- https://doi.org/10.1038/s44303-026-00187-2
- Primary Topic
- Advanced Fluorescence Microscopy Techniques
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Howard Hughes Medical Institute
- University of Pennsylvania
- Pennsylvania State University
- Children's Miracle Network Hospitals
- Consejo Nacional de Investigaciones Científicas y Técnicas
- Agencia Nacional de Promoción de la Investigación, el Desarrollo Tecnológico y la Innovación
- National Institutes of Health
- Penn State College of Medicine
- Medical Research Council