Analyte Detection Reagents Using Temporal Signal Signatures
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing multiplexing techniques in biology are limited by the number of available colors, typically restricted to 4 or 5, and methods like quantum dots or nanostrings face challenges in simultaneous detection of multiple analytes due to spatial resolution and sample requirements.
Innovation Solution
The use of detection reagents comprising probe reagents conjugated with nucleic acid labels that generate distinct signal signatures in a temporally-sequential manner, allowing for multiplexed detection and identification of analytes through a temporal series of optical imaging steps.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Device complexity
If traditional optical labeling with fluorophores or chromophores is used, then the detection method is simple and effective, but the number of detectable probes is limited to 4 or 5 colors
Solution Approach 1:
The detection system is segmented into multiple temporal stages, where each stage detects a subset of probes using a limited palette of fluorophores. By dividing the multiplexing task across time rather than space, the system achieves high probe multiplexing (100+ analytes) while maintaining simple optical detection at each stage.
Solution Approach 2:
The invention transitions from spatial multiplexing (using different colors simultaneously) to temporal multiplexing (using the same colors at different times). This dimensional change from spatial to temporal domain allows dramatically increased probe capacity without requiring additional fluorophores or complex imaging hardware.
2Quantity of substance
If quantum dots are used to increase the number of available colors, then the color range is expanded, but simultaneous detection of more than 6 colors is difficult to achieve
Solution Approach 1:
The system employs periodic action by cycling through multiple detection stages, where fluorophores are excited and detected in repeated temporal cycles. Each fluorophore is detected multiple times across different stages, allowing the same limited set of colors to encode information about many more probes through temporal patterning.
3Quantity of substance
If mixtures or ratio of fluorophores are used as new colors, then multiplexing is extended to hundreds of analytes, but the technology is limited to flow-cytometry based analyses due to label size
Solution Approach 1:
The invention extracts the temporal dimension from the detection process, separating the multiplexing function from the optical detection function. By taking out the temporal coding element and using it independently, the system achieves high multiplexing without requiring large microbead labels or flow-cytometry infrastructure, enabling application in fixed-cell imaging and other formats.
4Quantity of substance
If nanostrings are used for detection, then colorful barcodes are created, but very high-resolution imaging and special flow apparatus are required
Solution Approach 1:
The system uses temporal copying of fluorescence signals, where the same fluorophore signals are detected repeatedly across multiple stages and time points. This temporal copying creates a unique signature pattern for each probe, analogous to barcodes, but achieved through simple temporal measurement rather than complex spatial barcode imaging.
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
This approach significantly increases the number of detectable probes and analytes in a single assay, enabling high-throughput detection and identification of multiple targets without the limitations of traditional optical labeling.
Implementation Method 1
detecting in a temporally-sequential manner said plurality of the pre-determined subsequences of said detection reagents, wherein said detection of the subsequences each generates a signal signature corresponding to said subsequence
Data Source
Figure 1
Figure 2
Figure 3
AI summary
The inventions provided herein relate to detection reagents, compositions, methods, and kits comprising the detection reagents for use in detection, identification, and/or quantification of analytes in a sample. Such detection reagents and methods described herein allow multiplexing of many more labeled species in the same procedure than conventional methods, in which multiplexing is limited by the number of available and practically usable colors.