Photocleavable Mass-Tag Probes for Multiplex MSI Biomarker Mapping
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Solution Overview
Problem
Current methods for multiplexed tissue imaging, such as fluorescence microscopy, are limited in their ability to simultaneously detect multiple biomarkers due to spectral overlap, while mass spectrometric imaging (MSI) lacks the capability to target specific intact molecules like proteins and nucleic acids, restricting its multiplexing and multi-omics capabilities.
Innovation Solution
Development of photocleavable mass-tags (PC-MTs) attached to probes like antibodies and nucleic acids, enabling high-plex MSI through a fast and efficient photocleavable linker, allowing for simultaneous detection of multiple biomarkers and combining untargeted MSI with targeted macromolecule analysis on a single specimen.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescence microscopy is used for multiplexed tissue imaging, then biomarker detection capability is improved, but spectral overlap limits the number of simultaneously detectable biomarkers to 3-5
Solution Approach 1:
The patent replaces optical detection (fluorescence microscopy) with mass spectrometric detection. Instead of using fluorophores that emit light at different wavelengths, the invention uses mass tags with distinct mass-to-charge ratios that are detected by MSI. This substitution eliminates spectral overlap limitations and enables simultaneous detection of many more biomarkers (multiplexing capacity >8) while maintaining high detection precision through mass spectral resolution.
2Adaptability or versatility
If mass spectrometric imaging is used for tissue analysis, then untargeted molecular profiling capability is improved, but inability to target specific intact molecules like proteins and nucleic acids restricts its multiplexing and multi-omics capabilities
Solution Approach 1:
The patent introduces mass tags as intermediary molecules that bridge targeted and untargeted MSI approaches. These mass tags are attached to specific antibodies or nucleic acid probes that target intact molecules of interest, while the mass tags themselves enable detection by MSI. This intermediary approach allows the method to maintain the untargeted profiling capability of MSI while adding specific targeting capability for proteins, nucleic acids, and other macromolecules, thereby enabling both high multiplexing and multi-omics analysis.
3Ease of operation
If conventional IHC or ISH methods are used with fluorophore or chromogenic agents, then visualization of spatial distribution of biomolecules is achieved, but simultaneous determination of 5 or more biomarkers is limited
Solution Approach 1:
The patent substitutes fluorophore/chromogenic labeling with mass tag labeling on antibodies and nucleic acid probes. Instead of visualizing biomarkers through optical microscopy with limited color channels, the invention uses mass spectrometric imaging to detect multiple mass tags simultaneously based on their unique mass-to-charge ratios. This substitution maintains ease of operation through standard MSI protocols while dramatically increasing multiplexing capacity to detect 5 or more biomarkers simultaneously on a single tissue section.
4Adaptability or versatility
If cycling strategies with iterative staining and photobleaching are used to increase multiplexing, then number of detectable biomarkers is improved, but process complexity and labor requirements increase
Solution Approach 1:
The patent applies preliminary action by attaching multiple distinct mass tags to different antibodies or nucleic acid probes before applying them to the tissue section. Unlike cycling strategies that require iterative staining and photobleaching steps, this approach allows all targeted biomarkers to be visualized simultaneously in a single staining step followed by MSI analysis. The mass tags are pre-configured with unique mass signatures that enable multiplexed detection without requiring repeated processing cycles, thereby reducing process complexity and labor requirements while maintaining high multiplexing capability.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables highly multiplexed and multi-omic tissue imaging, overcoming limitations of existing methods by allowing simultaneous detection of a wide range of biomarkers in various tissues, including brain, tonsil, and breast cancer samples, with robust sensitivity and correlation with conventional fluorescence imaging.
Implementation Method 1
photocleavable linker...fast and efficient photocleavable linker
Data Source
AI summary
The field of this invention relates to immunohistochemistry (IHC) and in situ hybridization (ISH) for the targeted detection and mapping of biomolecules (e.g., proteins and miRNAs) in tissues or cells for example, for research use and for clinical use such by pathologists (e.g., biomarker analyses of a resected tumor or tumor biopsy). In particular, the use of mass spectrometric imaging (MSI) as a mode to detect and map the biomolecules in tissues or cells for example. More specifically, the field of this invention relates to photocleavable mass-tag reagents which are attached to probes such as antibodies and nucleic acids and used to achieve multiplex immunohistochemistry and in situ hybridization, with MSI as the mode of detection/readout. Probe types other than antibodies and nucleic acids are also covered in the field of invention, including but not limited to carbohydrate-binding proteins (e.g., lectins), receptors and ligands. Finally, the field of the invention also encompasses multi-omic MSI procedures, where MSI of photocleavable mass-tag probes is combined with other modes of MSI, such as direct label-free MSI of endogenous biomolecules from the biospecimen (e.g., tissue), whereby said biomolecules can be intact or digested (e.g., chemically digested or by enzyme).