Axially swept open-top light-sheet microscopy for densely labeled clinical specimens

Area of Science:

• Biomedical Imaging
• Microscopy Techniques
• Pathology

Background:

• Open-top light-sheet (OTLS) microscopy allows rapid 3D imaging of large, optically cleared specimens.
• This facilitates nondestructive 3D pathology, offering advantages over traditional histology like comprehensive sampling and 3D structure visualization.
• Clinical specimens are typically stained with broad-spectrum dyes, necessitating high-contrast imaging.

Purpose of the Study:

• To develop an OTLS microscope capable of high-resolution 3D imaging over a large field of view.
• To overcome the trade-off between optical sectioning and field of view in existing OTLS systems.
• To enable high-contrast 3D imaging of densely labeled clinical specimens.

Main Methods:

• Implementation of a voice-coil-based axial sweeping mechanism to improve optical sectioning.
• Utilizing a non-orthogonal dual-objective (NODO) architecture.
• Achieving a 10-mm working distance with reduced sensitivity to refractive index mismatches.

Main Results:

• The developed OTLS microscope achieves 2 µm axial resolution.
• Imaging is performed over a 750 × 375 µm field of view.
• High-contrast 3D imaging of clinical specimens was demonstrated.

Conclusions:

• The novel OTLS microscope design effectively addresses limitations in optical sectioning and field of view.
• This advancement supports high-contrast, nondestructive 3D pathology for clinical specimens.
• The NODO architecture provides a robust platform for advanced biomedical imaging.