Efficient 3D light-sheet imaging of very large-scale optically cleared human brain and prostate tissue samples

Area of Science:

• Biomedical Imaging
• Microscopy Techniques
• Histopathology

Background:

• 3D imaging of human tissues at cellular resolution and large fields of view (FOV) is crucial for research and clinical diagnostics.
• Existing methods often face limitations in scale, resolution, or speed when imaging large tissue volumes.

Purpose of the Study:

• To demonstrate the feasibility of light-sheet microscopy for imaging large-scale (cm³), cleared human tissue samples.
• To present a novel prototype, the cleared-tissue dual view Selective Plane Illumination Microscope (ct-dSPIM), for high-resolution 3D tissue imaging.
• To evaluate the imaging quality and potential clinical applications of the ct-dSPIM system.

Main Methods:

• Development of a cleared-tissue dual view Selective Plane Illumination Microscope (ct-dSPIM) prototype.
• Application of the FFPE-MASH protocol for tissue clearing.
• Utilized mosaic scanning techniques at varying resolutions (Mosaic 16, 4, and 0.5) for imaging formalin-fixed human brain and prostate cancer samples.

Main Results:

• Successfully imaged ~5 cm³ human brain and ~7 cm³ FFPE prostate cancer samples with cellular resolution.
• Demonstrated fast 3D acquisition speeds, ranging from ~1.7 h/cm³ (16.4 µm resolution) to ~15.8 h/cm³ (0.5 µm resolution).
• Visualized cortical layers and neurons in human brain tissue and confirmed suitability for Gleason score grading in prostate cancer samples.

Conclusions:

• ct-dSPIM is a powerful technique for quantitative 3D assessment of large-scale MASH-prepared human tissue samples.
• The system offers high-resolution imaging capabilities suitable for both fundamental biological investigations and clinical diagnostics.
• This technology holds considerable future potential for advancing histopathology and personalized medicine.