A multiwell plate approach to increase the sample throughput during tissue clearing

Area of Science:

• Biomedical Diagnostics
• Pathology
• Cellular Biology

Background:

• Traditional 2D pathology limits data quality.
• Current 3D tissue clearing workflows are complex and time-consuming.
• Increased throughput is needed for advanced biomedical diagnostics.

Purpose of the Study:

• To develop a high-throughput, parallel processing method for 3D tissue clearing and analysis.
• To reduce hands-on time and improve efficiency in 3D pathology workflows.
• To adapt existing clearing protocols for multi-sample compatibility.

Main Methods:

• A novel workflow combining a tissue clearing device with a six-well multiwell plate for parallel processing.
• Integration of passive clearing methods like Clear, Unobstructed Brain/Body Imaging Cocktails and Computational (CUBIC) analysis.
• Streamlined gel embedding, high-speed light-sheet microscopy, and cloud-based data analysis within the multiwell plate format.

Main Results:

• Achieved parallel tissue clearing, immunostaining, imaging, and analysis, increasing 3D pathology throughput.
• Reduced overall hands-on time per sample despite a slight increase in individual sample processing time.
• Demonstrated successful whole-organ analysis, completing clearing, imaging, and data analysis within 10 days for some samples.

Conclusions:

• The developed six-well plate-based parallel processing method significantly enhances the throughput of 3D pathology.
• This approach offers a practical and efficient platform for cellular biology and pathology research.
• The streamlined workflow facilitates advanced biomedical diagnostics and data acquisition.