Photocleavable Mass Tags for Targeted Multiplex MSI in Tissues
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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 ability to target specific intact molecules like proteins and nucleic acids, restricting its multiplexing capacity and multi-omics capabilities.
Innovation Solution
Development of photocleavable mass-tags (PC-MTs) attached to probes like antibodies and nucleic acids, allowing for high-plex MSI by facilitating the labeling of targeted macromolecules and combining with untargeted MSI for multi-omic tissue imaging on a single specimen.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
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 relying on optical properties that suffer from spectral overlap, the invention uses mass-to-charge ratio measurements to distinguish between different biomarkers, enabling detection of 10+ biomarkers simultaneously without spectral interference.
Solution Approach 2:
The invention changes the detection parameter from optical wavelength (where spectral overlap occurs) to mass-to-charge ratio (where each biomarker has a unique signature). This parameter transformation allows for high-plex detection by exploiting the higher dimensionality and resolution of mass spectrometry compared to optical spectroscopy.
2Quantity of substance
If mass spectrometric imaging is used for tissue imaging, then multiplexing capacity is improved, but ability to target specific intact molecules like proteins and nucleic acids deteriorates
Solution Approach 1:
The patent introduces photocleavable mass tags as intermediary molecules that bridge the gap between intact biomarkers and mass spectrometric detection. These tags are attached to antibodies or nucleic acid probes that specifically bind to target molecules, and are subsequently cleaved by light to release detectable mass signatures, enabling both specific targeting and high-plex detection.
Solution Approach 2:
The detection system is segmented into two functional components: (1) intact biomarker-probe complexes that maintain spatial information and specificity, and (2) cleaved mass tags that provide detectable mass signatures. This segmentation allows the system to preserve the advantages of both intact molecule detection and mass spectrometry-based multiplexing.
3Quantity of substance
If photocleavable mass-tags are attached to probes for high-plex MSI, then multiplexing capability is improved, but complexity of probe labeling process increases
Solution Approach 1:
The patent develops universal photocleavable mass tag platforms that can be attached to different types of probes (antibodies, nucleic acid probes, lectins) through standardized chemistry. This universal approach simplifies the labeling process by providing a single set of reagents and protocols that work across multiple probe types and target molecules, reducing overall system complexity despite enabling high-plex detection.
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 achieving robust sensitivity and allowing direct correlation with conventional fluorescence imaging, and performing both label-free untargeted small molecule and targeted macromolecular biomarker detection on the same tissue section.
Implementation Method 1
illumination with a UV light source (e.g., 365 nm) for 5-30 minutes to photocleave the mass-tags
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).