Microfluidic In-Pen Assays Using Reporter Diffusion Imaging

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Solution Overview

Problem

Existing technologies face challenges in accurately determining the quantity of analytes produced by micro-objects confined within microfluidic chambers, particularly due to sensitivity to the position of biological micro-objects and fluctuations in analyte concentration.

Innovation Solution

An imaging system and method that includes an image acquisition unit, an image processing unit, and a scoring engine to define an area of interest within microfluidic chambers, analyzing image areas sensitive to analyte concentration fluctuations and least sensitive to micro-object position, using soluble reporter molecules with detectable labels to quantify analyte secretion.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional imaging methods are used to measure analyte concentration in microfluidic chambers, then the measurement process is simple, but the measurement precision is poor due to sensitivity to micro-object position and analyte concentration fluctuations

Engineering Contradiction:

Improveanalyte concentration measurement precisionVSAvoidimaging system complexity

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The chamber is divided into multiple image areas, each associated with different micro-objects. By segmenting the measurement space and analyzing analyte concentration fluctuations in each segment independently, the system achieves more precise measurements while accounting for local variations in micro-object position and analyte distribution.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The system transitions from measuring absolute analyte concentration to measuring analyte concentration fluctuations over time. This temporal dimension provides additional information that enables more precise quantification of analyte production by micro-objects, overcoming limitations of spatial measurement alone.

Inventive Principle:

Principle #17Another dimension (Dimensionality change)

2Measurement precision

If the entire chamber area is used for imaging, then the field of view is maximized, but the measurement accuracy decreases due to interference from micro-object position variations

Engineering Contradiction:

Improveanalyte concentration measurement accuracyVSAvoidimaging area

Core Design Contradiction:

Measurement precisionVSArea of stationary object

Solution Approach 1:

Different image areas within the chamber are assigned different weights or levels of importance based on their sensitivity to analyte concentration fluctuations and insensitivity to micro-object position. By focusing analysis on locally optimized areas rather than treating the entire chamber uniformly, the system achieves higher measurement accuracy.

Inventive Principle:

Principle #3Local quality

Solution Approach 2:

The chamber imaging area is segmented into multiple image areas, each optimized for specific measurement purposes. This segmentation allows the system to exclude or de-emphasize areas where micro-objects are located, thereby improving overall measurement accuracy while maintaining comprehensive field of view.

Inventive Principle:

Principle #1Segmentation

3Productivity

If rapid imaging is performed to capture analyte concentration changes, then the productivity of analysis is improved, but the measurement precision may be compromised due to reduced integration time

Engineering Contradiction:

Improveanalysis throughputVSAvoidanalyte concentration measurement precision

Core Design Contradiction:

ProductivityVSMeasurement precision

Solution Approach 1:

The system performs periodic imaging at multiple time points to capture analyte concentration fluctuations. By analyzing the temporal pattern of these periodic measurements, the system can distinguish between signal variations caused by analyte production and those caused by noise or position drift, thereby maintaining precision while enabling rapid analysis.

Inventive Principle:

Principle #19Periodic action

Solution Approach 2:

The system establishes baseline measurements of analyte concentration at initial time points before significant changes occur. These preliminary measurements serve as reference values that enable more precise quantification of subsequent analyte production, allowing rapid imaging without sacrificing measurement accuracy.

Inventive Principle:

Principle #10Preliminary action

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 precise quantification of analyte production by micro-objects, facilitating clonal line development and secretion level assessment, with improved accuracy and sensitivity.

Implementation Method 1

a second fluidic medium into the flow region, wherein the second fluidic medium comprises a plurality of soluble reporter molecules

Methodology Applied

Scientific EffectDiffusion: Diffusion

Implementation Method 2

bind the secreted analyte and a detectable label

Methodology Applied

Scientific EffectBinding: Absorption (physical)

Implementation Method 3

detectable label

Methodology Applied

Scientific EffectLight detection: Light

Data Source

PatentUS20260056208A1Methods, Systems and Kits for In-Pen Assays

Publication Date: 2026.02.26 BRUKER SPATIAL BIOLOGY INC

AI summary

Methods, systems and kits are described herein for detecting the results of an assay. In particular, the methods, systems and devices of the present disclosure rely on a difference between the diffusion rates of a reporter molecule and an analyte of interest in order to quantify an amount of analyte in a microfluidic device. The analyte may be a secreted product of a biological micro-object.