Large-scale expanded sample imaging with tiling lattice lightsheet microscopy.

Area of Science:

• Biophysics
• Microscopy
• Cell Biology

Background:

• Observing biological nanostructures is crucial for understanding cellular functions.
• Expansion microscopy (ExM) allows super-resolution visualization of biological samples using conventional light microscopes.
• Physical expansion in ExM necessitates advanced deep and precise 3D imaging techniques.

Purpose of the Study:

• To explain the principles of tiling lattice lightsheet microscopy (tLLSM).
• To discuss the applications of tLLSM in conjunction with ExM.
• To highlight tLLSM's capability for high-precision 3D imaging of expanded biological samples.

Main Methods:

• Utilizes lattice lightsheet microscopy (LLSM) for planar illumination confined within high numerical aperture objectives.
• Incorporates optical tiling to expand the field of view (FoV) without mechanical sample or objective movement.
• Combines LLSM with optical tiling to create tLLSM for enhanced 3D imaging.

Main Results:

• LLSM provides deep and precise 3D imaging capabilities suitable for ExM.
• Optical tiling within LLSM effectively increases the field of view for comprehensive imaging.
• tLLSM integrates these features for advanced visualization of expanded biological samples.

Conclusions:

• tLLSM is a powerful technique for high-precision 3D imaging of samples prepared with ExM.
• This method addresses the increased imaging demands posed by physically expanded biological specimens.
• tLLSM facilitates a deeper understanding of biological nanostructures and their functions.