Efficient and cost-effective 3D cellular imaging by sub-voxel-resolving light-sheet add-on microscopy.

Area of Science:

• Biomedical imaging
• Microscopy techniques
• Optical engineering

Background:

• Light-sheet fluorescence microscopy (LSFM) offers high-speed, low phototoxicity live imaging.
• Commercial LSFM systems are often large and expensive, limiting accessibility.
• Conventional wide-field microscopes lack the resolution and speed for advanced 3D live imaging.

Purpose of the Study:

• To develop a cost-effective method for enhancing conventional microscopes for advanced light-sheet imaging.
• To achieve resolution-enhanced, fast 3D live imaging capabilities using an add-on device.
• To enable sub-voxel-resolving (SVR) imaging from existing wide-field microscope infrastructure.

Main Methods:

• Developed a miniature add-on device creating a horizontal laser light-sheet for optical sectioning.
• Implemented an off-axis scanning strategy combined with a sub-voxel-resolving (SVR) algorithm.
• Utilized sub-voxel spatial shifts for reconstructing high-resolution 3D image volumes.

Main Results:

• Achieved a twofold increase in imaging resolution compared to conventional methods.
• Successfully imaged muscle activity in C. elegans, zebrafish embryo heartbeats, and cleared mouse brains at high spatiotemporal resolution.
• Demonstrated the SLAM method's capability for fast 3D live imaging with super-resolved voxel capability.

Conclusions:

• The Sub-Voxel-Resolving (SVR) Light-sheet Add-on Microscopy (SLAM) method provides an efficient and affordable upgrade for conventional microscopes.
• SLAM enables fast, resolution-enhanced 3D live imaging, broadening access to advanced microscopy techniques.
• This technology facilitates detailed observation of biological processes in various model organisms and tissues.