Isotropic imaging across spatial scales with axially swept light-sheet microscopy.

Area of Science:

• Life Sciences
• Microscopy
• Biotechnology

Background:

• Light-sheet fluorescence microscopy is popular for its high resolution and gentle illumination.
• Existing techniques may have limitations in field of view, resolution, or optical sectioning.

Purpose of the Study:

• To provide a detailed protocol for assembling and operating axially swept light-sheet microscopy (ASLM).
• To democratize ASLM by offering practical guidance, schematics, and software for diverse biological imaging needs.

Main Methods:

• Detailed assembly and operation instructions for ASLM variants.
• Provision of part lists, schematics, and control software.
• Guidance on tuning imaging parameters for various sample types.

Main Results:

• ASLM achieves an unparalleled combination of field of view, optical resolution, and optical sectioning.
• The protocol enables imaging of molecular detail, subcellular organization, and whole-organism anatomy.
• ASLM can be adapted for chemically expanded, living, or cleared biological samples.

Conclusions:

• This protocol serves as a comprehensive guide for adopting ASLM in biological imaging.
• It empowers researchers to build and utilize custom imaging technology for advanced applications.
• Building an ASLM is feasible within 1-2 months, depending on user experience.