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

Area of Science:

• Biomedical Optics
• Microscopy
• Neuroscience Imaging

Background:

• Three-dimensional structured illumination microscopy (3D-SIM) enhances fluorescence imaging resolution and contrast but is limited in deep tissue imaging due to aberrations.
• Current 3D-SIM systems are typically inverted, restricting their use in live imaging applications requiring specimen manipulation.

Purpose of the Study:

• To develop a novel upright 3D-SIM system capable of deep tissue imaging.
• To overcome the limitations of sample-induced aberrations and improve contrast and resolution in thick biological samples.

Main Methods:

• Development of an upright 3D-SIM system (Deep3DSIM) incorporating adaptive optics for aberration correction.
• Implementation of remote focusing to enable volume imaging without moving the specimen or objective lens.
• Testing the system's performance on various biological samples, including the Drosophila brain.

Main Results:

• Deep3DSIM successfully corrects sample-induced aberrations, significantly improving image contrast and resolution.
• The system enables high-quality 3D-SIM imaging beyond 130 µm into the Drosophila brain, overcoming previous depth limitations.
• The upright configuration facilitates live imaging and specimen manipulation, expanding the applicability of 3D-SIM.

Conclusions:

• The developed Deep3DSIM system represents a significant advancement for deep tissue fluorescence imaging using 3D-SIM.
• Adaptive optics and remote focusing are crucial for overcoming aberrations and enabling high-performance 3D-SIM in challenging samples.
• This technology opens new avenues for studying complex biological structures in vivo.