Optical Clearing and Labeling for Light-sheet Fluorescence Microscopy in Large-scale Human Brain Imaging

Area of Science:

• Neuroscience
• Biotechnology
• Microscopy

Background:

• Processing postmortem human brains for high-resolution imaging is challenging due to size and complexity.
• Existing clearing techniques struggle with the intricate structure of the human brain.

Purpose of the Study:

• To present a novel protocol for efficient 3D reconstruction of human brain volumes.
• To enable high-resolution imaging of large brain sections.

Main Methods:

• The SWITCH - H2O2 - Antigen Retrieval - 2,2'-thiodiethanol (TDE) [SHORT] tissue transformation protocol was employed.
• Simultaneous processing of multiple thick brain sections for clearing and multi-labeling.
• Light-sheet fluorescence microscopy (LSFM) for sequential imaging of cleared slices.

Main Results:

• The SHORT protocol achieved rapid and homogeneous clearing and multi-labeling of thick brain slices.
• Neuronal subpopulations in both white and grey matter were identified.
• High-resolution, multi-channel imaging allowed for rapid 3D reconstruction of large volumetric areas.

Conclusions:

• The combined SHORT protocol and LSFM provide a high-throughput method for comprehensive human brain structural characterization.
• This approach significantly reduces the time required for high-resolution 3D brain reconstruction.
• Enables detailed analysis of cytoarchitecture in large brain volumes.