Sequential Biological Sample Analysis for Multiplexed Target Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for detecting multiple targets in a biological sample are limited by sensitivity, accuracy, and multiplexing capabilities, often allowing detection of only a few targets at a time due to limitations in fluorescence-based detection systems, necessitating additional samples for comprehensive analysis.
Innovation Solution
The method involves contacting a biological sample with multiple target-binding probes to form target-bound probes, observing initial signals, modifying them, and generating secondary signals through sequential detection using signal-generating probes, allowing for the detection of a plurality of targets in a single sample using a device with a sample handling, reagent dispensing, and signal detection system.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescence-based detection systems are used to detect multiple targets, then detection sensitivity is improved, but the number of targets that can be accurately detected is limited to only a few at a time due to signal overlap
Solution Approach 1:
The detection process is divided into multiple sequential stages. First, multiple target-binding probes are applied simultaneously to bind to different targets. Then, signal-generating probes are applied in sequential rounds, with each round detecting a specific subset of targets. After each detection round, the observed signals are modified (e.g., through photobleaching or chemical quenching) before the next round begins. This segmentation allows many more targets to be detected than would be possible in a single simultaneous measurement, resolving the contradiction between detection sensitivity and multiplexing capability.
Solution Approach 2:
The detection system employs periodic action by conducting multiple detection rounds sequentially. Each round involves applying signal-generating probes, detecting signals, modifying the signals, and then proceeding to the next round. This periodic cycle of detection-modification-detection enables the system to accumulate information about many different targets over time, overcoming the limitation of detecting only a few targets simultaneously while maintaining high detection sensitivity through repeated measurements.
2Adaptability or versatility
If multiple biological samples are analyzed to detect all relevant targets, then comprehensive target detection is achieved, but the ability to accurately determine relative characteristics of targets is limited
Solution Approach 1:
The invention merges multiple detection capabilities into a single biological sample through sequential detection rounds. By applying different signal-generating probes in successive rounds to the same sample, the system simultaneously detects multiple targets and preserves their spatial relationships and relative concentrations. This merging approach eliminates the need to analyze separate samples, allowing comprehensive target detection while maintaining accurate determination of relative characteristics within the single sample context.
Solution Approach 2:
The method creates multiple detection 'copies' through sequential rounds of signal generation and detection. Each round generates a set of signals corresponding to specific targets, and these signal sets are accumulated and integrated. This copying process allows the system to extract information about multiple targets from the same sample multiple times, achieving comprehensive detection while preserving the ability to compare relative characteristics across all detected targets.
3Adaptability or versatility
If additional biological samples are used to detect more targets, then target coverage is improved, but sample availability becomes a limiting factor
Solution Approach 1:
The detection system maintains continuity of useful action by performing multiple detection rounds on the same biological sample without requiring additional samples. Each round of signal generation and detection continues to extract information from the original sample, accumulating comprehensive target coverage over time. This continuous utilization of the single sample overcomes the limitation of sample availability, allowing detection of many more targets than would be possible with a single simultaneous measurement while preserving the limited sample material.
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 enables accurate detection of multiple targets in a single biological sample, overcoming limitations in sensitivity and multiplexing, and allowing for detailed analysis of spatial distribution and concentration, even with limited sample amounts.
Implementation Method 1
contacting the biological sample with a plurality of target-binding probes to form a plurality of target-bound probes
Implementation Method 2
generating a first set of signals from a first set of the plurality of target-bound probes
Data Source
AI summary
Methods for detecting a plurality of targets in a biological sample are provided. The method comprises contacting the biological sample with a plurality of target-binding probes simultaneously to form a plurality of target-bound probes and observing the signals from the target-bound probes sequentially. An associated kit and device for detection of the plurality of targets are also provided.