Miniscope3D: optimized single-shot miniature 3D fluorescence microscopy

Area of Science:

• Biomedical Engineering
• Optical Microscopy
• Systems Biology

Background:

• Conventional miniature microscopes offer 2D imaging, limiting 3D biological studies.
• Existing 3D miniature microscopes are bulky, heavy, and have limited resolution.
• There is a need for compact, high-resolution 3D miniature microscopes.

Purpose of the Study:

• To develop a compact, lightweight miniature microscope capable of high-resolution 3D fluorescence imaging.
• To overcome the limitations of existing 2D and 3D miniature microscopy systems.
• To enable advanced biological research requiring volumetric imaging in constrained environments.

Main Methods:

• Replaced the tube lens of a 2D Miniscope with an optimized multifocal phase mask at the objective's aperture stop.
• Encoded 3D fluorescence intensity into a single 2D measurement, with 3D volume recovered via a sparsity-constrained inverse problem.
• Developed methods for phase mask design, fabrication, and an efficient forward model accounting for aberrations.

Main Results:

• Achieved a prototype microscope measuring 17 mm tall and weighing 2.5 grams.
• Obtained 2.76 μm lateral and 15 μm axial resolution across a 900 × 700 × 390 μm³ volume at 40 volumes/second.
• Demonstrated superior performance over existing systems, with 2x better resolution and 10x larger depth range.

Conclusions:

• The novel multifocal phase mask design enables compact, high-resolution single-shot 3D fluorescence imaging.
• This technology significantly advances miniature microscopy for applications like neural imaging and dynamic sample studies.
• The system offers a smaller, lighter, and more capable alternative for in-situ 3D imaging.