Efficient Tissue Clearing and Multi-Organ Volumetric Imaging Enable Quantitative Visualization of Sparse Immune Cell...

Area of Science:

• Cellular and Molecular Biology
• Biomedical Imaging
• Pathophysiology

Background:

• Understanding cellular spatial information within tissue microenvironments is crucial for elucidating complex pathophysiological processes.
• Current organ-specific tissue clearing and volumetric imaging protocols are often complex, costly, and labor-intensive.
• There is a need for simplified, broadly applicable methods for high-resolution 3D tissue analysis.

Purpose of the Study:

• To develop a simplified, cost-effective, and non-hazardous method for efficient tissue clearing and multi-organ volumetric imaging.
• To enable multiplexed immunolabeling and high-resolution imaging of cellular structures in cleared tissues.
• To demonstrate the utility of the developed method in analyzing cellular events in disease models.

Main Methods:

• Developed EMOVI (efficient tissue clearing and multi-organ volumetric imaging), a streamlined protocol for tissue clearing.
• Utilized multiplexed antibody-based immunolabeling for specific cellular and molecular detection.
• Employed widefield microscopy with real-time computational clearing for image acquisition.

Main Results:

• EMOVI achieved adequate tissue transparency while preserving cellular morphology and fluorochromes.
• The method successfully enabled the detection and quantification of scarce cell populations in pneumonitis models.
• EMOVI facilitated histo-cytometric analysis of MHC-II expressing cells in nephritic kidneys, revealing distinct spatial distributions.
• Real-time computational clearing with widefield microscopy proved effective for rapid imaging and rare event detection.

Conclusions:

• EMOVI offers a flexible, cost-effective, and accessible approach to tissue clearing and volumetric imaging.
• The technique supports multiplexed immunolabeling and analysis of immune cells in various organs and disease contexts.
• EMOVI has the potential to broaden the application of volumetric imaging in biological and medical research.