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Techniques for visualizing the three-dimensional structure of biological tissue in the brain are particularly important in understanding the pathological processes of neurological diseases such as Parkinson’s disease and Alzheimer’s disease. However, current techniques that allow full visualization of brain structures are not widely used and are expensive.
A team from the Margaret KL Chan Research Center for Parkinsonism Management at the Chinese University of Hong Kong (CUHK) 3D immunostaining Technology – ThICK (Thermal Immunohistochemistry with Optimized Kinetics) staining technology. Generate stabilized antibodies for rapid labeling and imaging of molecules in living tissue and visualize their 3D structures in detail. The research results are It was published As the cover story for the latest issue of the international scientific journal Nature Methods.
Widely used to visualize the structure of living tissue, immunostaining uses antibodies to target biomolecules within the tissue to reveal tissue structure.
Dr. Lai Haimin, assistant professor of psychiatry at CUHK School of Medicine and principal investigator at the Margaret KL Chang Parkinsonism Management and Research Center, pointed out that antibodies are usually sensitive to heat and chemicals. Denaturant.
When used in immunostaining, these properties limit the choice of temperature and denaturant as the antibody can become unstable and lose functionality. The inability of antibodies to reach deep into tissues hinders their application to high-throughput 3D structural studies.
The team has developed a chemical technique that can transform antibodies commonly used in research, making them much more resistant to heat and denaturants. A thermostable antibody called SPEAR (Synergistically Protected Polyepoxide Cross-Linked Fab Conjugated Antibody Reagent) can be used to achieve immunolabeling of biomolecules deep within intact tissue.
Therefore, the use of SPEAR with ThICK staining can help overcome the limitations of deep 3D immunostaining and optimize visualization of brain structures.
Dr. Zachary Lau, an investigator at the Margaret KL Chan Research Center for the Management of Parkinsonism, said ThICK staining achieves whole-organ immunolabeling more quickly compared to other advanced techniques. 1 to 3 days is a very short time.
The research team used SPEAR to implement the ThICK staining technique, and the results showed that immunolabeling of the entire mouse brain could be achieved within 72 hours. When applied to human brain tissue, one-third the antibody can achieve almost four-fold greater penetration.
The research team said the new method is compatible with the wide range of antibodies available, as well as different classes of tissue preservation and removal methods. Therefore, it can be easily processed and implemented in most laboratories. This new technique has particular application in the study of neurodegenerative diseases.
The team hopes their research can accelerate the path to new tools for diagnosing and treating disease.
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