Microfluidic In-Pen Assay Imaging for Precise Analyte Quantification
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Solution Overview
Problem
Existing technologies face challenges in accurately determining the quantity of an analyte produced by a micro-object confined within a chamber in a microfluidic device, 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 the chamber, which is sensitive to analyte concentration fluctuations and least sensitive to the position of biological micro-objects, allowing for precise quantification of analyte production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If imaging is performed across the entire chamber, then complete coverage is achieved, but sensitivity to positional fluctuations of biological micro-objects increases
Solution Approach 1:
The patent applies local quality by defining a specific area of interest within the chamber that has optimized properties for measurement. This area is positioned to minimize sensitivity to micro-object position while maximizing analyte concentration detection capability, thereby resolving the contradiction between complete coverage and positional sensitivity.
Solution Approach 2:
The imaging area is segmented into an area of interest and other regions. By focusing measurement resources on the optimized area of interest rather than the entire chamber, the system achieves precise analyte measurement while reducing the impact of positional variations of biological micro-objects.
2Measurement precision
If the area of interest is optimized for analyte concentration sensitivity, then measurement precision improves, but the area covers less of the chamber
Solution Approach 1:
The patent applies partial action by focusing imaging resources on a specific area of interest rather than attempting to image the entire chamber. This concentrated approach on the optimized region achieves superior measurement precision for analyte concentration, accepting that not the entire chamber is imaged with the same detail.
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 accurate and reliable determination of analyte quantity by minimizing positional sensitivity and enhancing measurement precision, facilitating clonal line development and secretion level assessment.
Implementation Method 1
allowing a portion of the plurality of soluble reporter molecules to diffuse into the chamber and bind to the analyte secreted therein
Implementation Method 2
each soluble reporter molecule includes a binding component configured to bind the secreted analyte
Data Source
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.