Spherical aberration correction for air objectives in light sheet microscopy

This technology is a hybrid solid-liquid optical system that corrects spherical aberration in low-cost air objectives, enabling high-resolution light sheet imaging of large cleared and living biological samples.

Unmet Need: Affordable subcellular-resolution 3D imaging of large samples

Light sheet microscopy is the method of choice for volumetric imaging of cleared and living samples, but achieving subcellular resolution depends on specialized immersion objectives. These objectives cost tens of thousands of dollars and offer only millimeters of working distance, limiting imaging depth and flexibility across sample types and immersion media. Inexpensive air objectives provide the long working distances needed for large specimens, but refraction at the air-liquid boundary causes severe spherical aberration that degrades lateral and axial resolution. As a result, routine high-throughput, high-resolution imaging of large intact specimens remains out of reach for most laboratories.

The Technology: Immersion-grade resolution from inexpensive long-working-distance air objectives

This technology pairs a solid optical element with a refractive index-matched liquid to pre-compensate the aberrations that arise when an air objective images into an immersion medium. A curved off-the-shelf lens serves as the sample chamber window and is positioned so that light rays cross the air-liquid boundary nearly perpendicular, suppressing spherical aberration. Matching the refractive index of the lens material to the chamber liquid further raises the objective’s effective numerical aperture. The approach applies to both illumination and detection paths, is objective-agnostic, and integrates with existing light sheet platforms without specialized optics or adaptive correction hardware.

The corrective optics and imaging systems have been prototyped and validated with cleared tissue and large intact biological specimens, achieving submicron resolution with standard air objectives.

Applications:

• High-resolution 3D imaging of cleared tissue
• High-content imaging for drug discovery and phenotypic screening
• 3D histopathology and digital pathology
• Research tool for neuroscience, developmental biology, and organoid analysis
• Manufacturing of corrective optical components and add-on modules for existing light sheet platforms

Advantages:

• Achieves submicron resolution with inexpensive long-working-distance air objectives
• Avoids the high cost and short working distance of immersion objectives
• Compatible with multiple immersion media and clearing protocols
• Objective-agnostic design integrates with existing light sheet platforms
• Uses simple off-the-shelf optical components, avoiding complex adaptive correction hardware

Lead Inventor:

Raju Tomer, Ph.D.

Patent Information:

Patent pending

Related Publications:

• Gong C, Affatato P, Fay M, Guttikonda SR, O’Connor NJ, Noble E, Heal M, Haydock B, Mapa R, De La Cruz ED, Gattoni G, Kowalko JE, Tosches MA, Gerfen CR, Hen R, Makinson CD, Hibshoosh H, Glaser JR, Tomer R. “Hybrid solid−liquid optics enable scalable, high-resolution light-sheet microscopy across diverse immersion media” Nat Biotechnol. 2026 Jun.

• Chen Y, Chauhan S, Gong C, Dayton H, Xu C, De La Cruz ED, Tsai YW, Datta MS, Rosoklija GB, Dwork AJ, Mann JJ, Boldrini M, Leong KW, Dietrich LEP, Tomer R. “Low-cost and scalable projected light-sheet microscopy for the high-resolution imaging of cleared tissue and living samples” Nat Biomed Eng. 2024 Sep; 8(9): 1109-1123.

• Migliori B, Datta MS, Dupre C, Apak MC, Asano S, Gao R, Boyden ES, Hermanson O, Yuste R, Tomer R. “Light sheet theta microscopy for rapid high-resolution imaging of large biological samples” BMC Biol. 2018 May 29; 16(1): 57.

Tech Ventures Reference:

• IR CU25135, CU25425, and CU26183

• Licensing Contact: Kristin Neuman