Whole-mouse immunolabeling at cellular resolution for comprehensive 3D atlases

Area of Science:

• Biological imaging
• Systems biology
• Immunohistochemistry

Background:

• Mapping complex biological systems and tracking disease progression at high resolution across the entire mammalian body is challenging.
• Existing methods lack the resolution or scope for comprehensive whole-body analysis.

Purpose of the Study:

• To present wildDISCO, a comprehensive protocol for whole-body immunolabeling, optical clearing, and imaging of mice at cellular resolution.
• To enable deep antibody penetration and detailed mapping of biological systems and disease models.

Main Methods:

• Developed wildDISCO (immunolabeling of wild-type mice and DISCO clearing) protocol.
• Optimized tissue permeabilization using cyclodextrin for enhanced antibody penetration.
• Detailed procedures for sample preparation, decolorization, decalcification, immunostaining, clearing, and 3D imaging.

Main Results:

• Achieved cellular resolution imaging across the entire mouse body.
• Enabled comprehensive mapping of neuronal, vascular, lymphatic, and immune systems.
• Facilitated systemic studies in disease models, including cancer and microbiome-host interactions.

Conclusions:

• wildDISCO is a broadly applicable platform for generating whole-body cellular atlases.
• The protocol enables systems-level investigations of health and disease.
• Requires standard immunohistochemistry facilities; initial training may be beneficial.