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A protocol to visualize on-target specific drug binding in mammalian tissue with cellular resolution using tissue
1Department of Neuroscience, The Scripps Research Institute, La Jolla, CA 92037, USA.
Abstract:
Here, we provide a protocol to visualize on-target specific drug binding in mammalian tissue with cellular resolution. By combining tissue clearing and click chemistry, this protocol allows fluorescence tagging of covalent drug binding in situ. In addition, the protocol is compatible with molecular marker staining for cell type identifications. For complete details on the use and execution of this protocol, please refer to Pang et al. (2022).
Insights
This study presents a new protocol for visualizing drug binding in tissues. The method uses tissue clearing and click chemistry to tag covalent drug binding with fluorescence at the cellular level.
Area of Science:
- Biomedical imaging
- Chemical biology
- Drug discovery
Background:
- Visualizing drug-target interactions in tissues is crucial for understanding drug efficacy and safety.
- Existing methods often lack the resolution or specificity to observe drug binding at the cellular level *in situ*.
Purpose of the Study:
- To develop a protocol for visualizing on-target, specific drug binding in mammalian tissues with single-cell resolution.
- To enable the direct observation of covalent drug interactions within their native biological context.
Main Methods:
- The protocol combines advanced tissue clearing techniques with bioorthogonal click chemistry.
- Fluorescence tagging is employed to visualize covalent drug binding *in situ*.
- Compatibility with molecular marker staining allows for cell type identification.
Main Results:
- Successful visualization of on-target drug binding at the cellular level in mammalian tissue.
- The protocol allows for specific tagging of covalent drug interactions.
- Integration with cell-specific markers enables co-localization studies.
Conclusions:
- This protocol provides a powerful tool for studying drug-target engagement in complex biological systems.
- It facilitates a deeper understanding of drug pharmacology and mechanism of action.
- The method is adaptable for various drug modalities and tissue types.
