Digital Assay Compartment Optimization for Concentration Precision
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Solution Overview
Problem
Current analytical assays for determining analyte particle concentrations face challenges in achieving high precision and dynamic range without requiring external calibration, often necessitating large numbers of compartments and complex equipment.
Innovation Solution
A method that divides analyte particles into a reduced number of compartments, assigning digital and analogue values based on signal measurements to estimate concentration, allowing for adjustable precision and dynamic range without external calibration, using a sample holder with a predetermined number of compartments optimized for the required precision and dynamic range.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If digital assays partition the sample into a large number of compartments to achieve high precision and wide dynamic range, then measurement precision and dynamic range are improved, but device complexity and the number of compartments required increase significantly
Solution Approach 1:
The invention partitions the sample into a smaller number of compartments compared to conventional digital assays. By optimizing the compartment number and volume, the patent achieves high measurement precision without requiring the large number of compartments (e.g., 200,000 or 600,000) needed in prior art digital PCR procedures.
Solution Approach 2:
The patent varies the volume of compartments as a key parameter to improve both precision and dynamic range. By using compartments of different volumes or optimizing the volume distribution, the assay can accurately measure concentrations across a wide dynamic range (spanning 1 million or more) while using fewer compartments than conventional approaches.
2Productivity
If analogue PCR techniques are used for concentration assessment, then the assay is quick and robust, but external calibration is required frequently and accuracy cannot be assessed at the point of measurement
Solution Approach 1:
The digital assay is self-calibrating and does not require external calibration standards. By partitioning the sample and using statistical analysis of the digital signals from multiple compartments, the assay independently determines the analyte concentration without relying on external reference samples or frequent calibration procedures.
3Adaptability or versatility
If the dynamic range is increased to span large concentration variations, then the assay becomes more versatile, but the number of compartments required increases proportionally
Solution Approach 1:
The patent uses variable compartment volumes as a key parameter to achieve wide dynamic range with fewer compartments. By optimizing the volume distribution and using statistical methods to analyze signals from compartments of different sizes, the assay can measure concentrations spanning 1 million or more fold variations without requiring proportionally large numbers of compartments.
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 and precise determination of analyte particle concentrations with reduced compartment numbers, eliminating the need for external calibration and simplifying the assay process, while maintaining a wide dynamic range.
Implementation Method 1
the signal generally increases in time in a reproducible manner, so that it is possible to define a threshold value of the intensity of fluorescence from the inspected volume
Data Source
AI summary
The present invention relates to a method for determining an estimate of a concentration of analyte particles E(C) as well as an apparatus for use in the inventive method, uses of the inventive method or the inventive apparatus, a sample holder and a kit for use in the inventive method.