Deep3DSIM: super-resolution imaging of thick tissue using 3D structured illumination with adaptive optics

Journal article

Three-dimensional structured illumination microscopy (3D-SIM) doubles the resolution of fluorescence imaging in all directions and enables optical sectioning with increased image contrast. However, 3D-SIM has not been widely applied to imaging deep in thick tissues due to its sensitivity to sample-induced aberrations, making the method difficult to apply beyond 10 µm in depth. Furthermore, 3D-SIM has not been available in an upright configuration, limiting its use for live imaging while manipulating the specimen, for example, with electrophysiology. Here, we have overcome these barriers by developing a novel upright 3D-SIM system (termed Deep3DSIM) that incorporates adaptive optics for aberration correction and remote focusing, reducing artefacts, improving contrast, restoring resolution, and eliminating the need to move the specimen or the objective lens in volume imaging. These advantages are equally applicable to inverted 3D-SIM systems. We demonstrate high-quality 3D-SIM imaging in various samples, including imaging more than 130 µm into the Drosophila brain.

Authors: Wang, J; Stoychev, D; Phillips, MA; Pinto, DMS; Parton, RM

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: eLife Sciences Publications
• Journal: eLife
• Volume: 14
• Article number: e102144
• Place of publication: England
• Publication date: 2025-10-28
• Acceptance date: 2025-09-04
• DOI: 10.7554/eLife.102144
• EISSN: 2050-084X
• Language: English