Multiplex Digital Target Detection Using Low-Occupancy 3D Partitions

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Solution Overview

Problem

Existing systems for high dynamic range digital counting of targets are limited by partition number, format, high apparatus costs, statistical error correction, material costs, insufficient precision and accuracy, low speed performance, and inability to preserve samples for further analysis.

Innovation Solution

Systems and methods for high dynamic range digital detection and quantitation of targets using a high number of partitions with low occupancy, enabling accurate and precise counting across a wide dynamic range, utilizing 3D imaging techniques and generating emulsions with gel-like properties for stable droplet detection.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If traditional partition-based systems are used for digital PCR, then target detection is achieved, but the number of partitions is limited and apparatus costs are high

Engineering Contradiction:

Improvenumber of partitionsVSAvoidapparatus costs

Core Design Contradiction:

Quantity of substanceVSDevice complexity

Solution Approach 1:

The system segments the detection problem into two independent components: (1) digital partitioning of the sample into many small reaction units using microfluidic droplet generation, and (2) parallel optical detection of multiple targets across all partitions simultaneously using spectral imaging. This segmentation enables thousands of partitions to be created without proportionally increasing apparatus complexity, as the detection step processes all partitions in parallel rather than sequentially.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The optical detection system is designed with multi-functionality to detect multiple targets simultaneously across all partitions using spectral unmixing and fluorescence lifetime imaging. A single imaging system performs what would traditionally require multiple separate detection channels, reducing apparatus costs while enabling detection of many targets across thousands of partitions.

Inventive Principle:

Principle #6Universality (Multi-functionality)

2Reliability

If high occupancy regime is used for target detection, then detection sensitivity is improved, but statistical error correction is required reducing accuracy

Engineering Contradiction:

Improvedetection sensitivityVSAvoidcounting accuracy

Core Design Contradiction:

ReliabilityVSMeasurement precision

Solution Approach 1:

The system changes the occupancy parameter from high to low (average less than 0.5 targets per partition) to eliminate the need for statistical error correction. This parameter change is compensated by increasing the total number of partitions and using parallel detection across all partitions, maintaining detection sensitivity while achieving Poisson-distribution-based counting accuracy without complex correction factors.

Inventive Principle:

Principle #35Parameter changes

3Productivity

If traditional detection methods are used, then target counting is achieved, but speed performance is low and samples cannot be preserved

Engineering Contradiction:

Improvecounting speedVSAvoidsample preservation

Core Design Contradiction:

ProductivityVSDuration of action of stationary object

Solution Approach 1:

The system replaces traditional sequential mechanical or chemical detection methods with parallel optical detection using spectral imaging and fluorescence lifetime imaging. This substitution enables simultaneous detection of multiple targets across thousands of partitions in a single measurement, dramatically increasing counting speed while preserving the sample in a closed microfluidic system for potential reuse or further analysis.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

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

Achieves accurate and precise counting of targets across a dynamic range of up to eight logarithms with high speed and low error, allowing for efficient multiplexed detection and quantitation of targets with improved precision and reduced costs.

Implementation Method 1

obtaining a fluorescence emission spectrum for each partition

Methodology Applied

Scientific EffectFluorescence: Fluorescence

Implementation Method 2

obtaining a fluorescence lifetime image of the set of partitions

Methodology Applied

Scientific EffectFluorescence lifetime: Fluorescence

Data Source

PatentUS20260002210A1Detection and digital quantitation of multiple targets

Publication Date: 2026.01.01 COUNTABLE LABS INC

AI summary

The disclosure provides compositions, methods, and systems for implementation of high-performance molecular diagnostic assays involving color combinatorics, stimulus-responsive probes, tandem probes, conjugated polymer probes, and other mechanisms for increasing the number of targets that can be simultaneously detected in a digital assay. Multiplexed detection of targets is achieved in a rapid manner, with respect to sample partitioning and target detection using multiple color channels for detection.