High-resolution in-depth imaging of optically cleared thick samples using an adaptive SPIM

Area of Science:

• Biomedical Optics
• Microscopy
• Biotechnology

Background:

• Light Sheet Fluorescence Microscopy (LSFM) enables deep tissue imaging but is limited by scattering and aberrations.
• Optical clearing methods homogenize refractive index (RI) to reduce scattering but introduce RI mismatches with imaging media.
• RI mismatches cause aberrations, degrading image quality in cleared biological samples.

Purpose of the Study:

• To develop an optical-chemical method to compensate for RI mismatches in optically cleared samples.
• To enable high-resolution, in-depth imaging of complex biological samples using LSFM.

Main Methods:

• An adaptive SPIM (Scanning Light Sheet Fluorescence Microscopy) system was employed.
• A water-based clearing protocol was utilized to homogenize sample RI.
• The combined method compensates for aberrations induced by RI mismatches.

Main Results:

• Successfully compensated for aberrations caused by RI mismatches in cleared samples.
• Achieved high-resolution, in-depth imaging of optically cleared complex thick samples.
• Demonstrated improved image quality compared to standard LSFM on cleared samples.

Conclusions:

• The developed optical-chemical method effectively corrects aberrations from RI mismatches.
• This approach enhances the capability of LSFM for imaging complex, cleared biological specimens.
• The technique holds promise for advanced in-depth imaging in biological research.