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Sample Preparation of Optically Cleared Liver Tissue to Identify Liver Macrophages Using 3D Microscopy
Otto Strauss1, Niklas K Björkström2
1Department of Medicine Huddinge, Center for Infectious Medicine, ANA Futura, Karolinska Institutet, Karolinska University Hospital Huddinge, Stockholm, Sweden. olstrauss@gmail.com.
Methods in Molecular Biology (Clifton, N.J.)
|July 2, 2020
Summary
Histology requires advanced imaging for liver macrophages. Clearing-enhanced 3D (Ce3D) and FUnGI methods enable 3D imaging of thick liver tissues for macrophage distribution analysis.
Area of Science:
- Histology
- Cell Biology
- Microscopy
Background:
- Histology studies tissue microanatomy and cell function within organs.
- Liver macrophages have complex shapes, necessitating thick tissue sections (>100 μm) for accurate distribution analysis.
- Standard tissue sections (5-10 μm) are insufficient for studying liver microanatomy and macrophage distribution.
Purpose of the Study:
- To describe methods for optically clearing thick liver tissue specimens.
- To enable three-dimensional (3D) imaging and analysis of macrophage distribution in the liver.
- To adapt clearing-enhanced 3D (Ce3D) and FUnGI techniques for liver tissue.
Main Methods:
- Optical clearing of liver tissue specimens thicker than 100 μm.
- Utilizing modified clearing-enhanced 3D (Ce3D) and FUnGI protocols.
- Employing antibody staining and refractive index matching to reduce light diffraction.
- Confocal microscopy for 3D imaging of cleared tissues.
Main Results:
- Successful optical clearing of liver tissue specimens exceeding 100 μm thickness.
- Demonstration of techniques to overcome light diffraction in opaque liver tissue.
- Enabled visualization of macrophage distribution in three dimensions within the liver microanatomy.
Conclusions:
- Ce3D and FUnGI are effective methods for optically clearing thick liver tissue.
- These techniques facilitate detailed 3D analysis of macrophage distribution in liver microanatomy.
- Advanced microscopy and clearing methods are crucial for studying complex tissue structures.
