Protocol for Imaging and Analysis of Mouse Tumor Models with CUBIC Tissue Clearing

Area of Science:

• Biotechnology
• Cancer Research
• Microscopy

Background:

• Tissue-clearing technologies have advanced significantly, particularly in neuroscience.
• CUBIC (Clear, Unobstructed Brain/Body Imaging with Clarity) has shown utility in cancer research.
• Previous work demonstrated CUBIC's potential for detecting cancer metastasis at single-cell resolution.

Purpose of the Study:

• To provide a detailed protocol for applying CUBIC tissue clearing in cancer research.
• To enable high-quality 3D imaging of tumors and metastasis at the whole organ/body level.
• To facilitate post-immunohistochemistry analysis following CUBIC clearing.

Main Methods:

• Detailed step-by-step protocol for CUBIC tissue clearing.
• Methodology for capturing high-resolution 3D images of cleared tissues.
• Procedure for performing immunohistochemistry on cleared samples.

Main Results:

• The CUBIC method allows for the detection of cancer metastasis with single-cell resolution.
• Analysis can be performed on whole mouse bodies or organs, providing a comprehensive view.
• High-quality 3D images reveal detailed tumor characteristics.

Conclusions:

• The CUBIC protocol is a valuable tool for detailed cancer metastasis and tumor analysis.
• This method supports single-cell resolution imaging in whole biological specimens.
• The protocol facilitates integrated tissue clearing, 3D imaging, and immunohistochemistry workflows.