Single-scan volumetric imaging throughout thick tissue specimens by one-touch installable light-needle creating...

Area of Science:

• Neuroscience
• Biomedical Imaging
• Cell Biology

Background:

• Dynamic changes in biological tissues necessitate 3D imaging with single-cell resolution.
• Existing volumetric imaging technologies often involve complex setups and require specialized expertise, posing a high barrier for biologists.
• There is a need for accessible and user-friendly tools for high-resolution 3D biological imaging.

Purpose of the Study:

• To develop an easy-to-use device for enhancing conventional two-photon microscopy systems.
• To enable simultaneous excitation and imaging of fluorophores throughout thick biological specimens.
• To facilitate high-resolution 3D visualization and monitoring of biological network activities.

Main Methods:

• Development of a novel light-needle creating device compatible with standard filter cube installation in fluorescent microscopes.
• Utilized the device with a conventional two-photon microscopy system for single-scan volumetric imaging.
• Applied the system to visualize 3D structures of YFP-expressing brain slices and detect calcium activity in mouse cortical slices.

Main Results:

• The light-needle device enabled simultaneous excitation of fluorophores across specimens over 200 μm thick with a single scan.
• Successfully visualized the 3D structure of transparent YFP-expressing brain slices.
• Detected calcium activities in acute mouse cortical slices (approx. 250 μm thick) with a temporal resolution of 7.5 Hz in neuronal populations.

Conclusions:

• The developed light-needle device significantly lowers the threshold for performing volumetric imaging in biological research.
• This technology offers a simplified approach for simultaneous 3D imaging of large biological tissues using conventional microscopy.
• The system demonstrates potential for advancing the study of dynamic cellular activities in complex biological networks.