Fluorescence Polarization Measuring Method for Wide Concentration Range
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Solution Overview
Problem
Fluorescence polarization methods face limitations in achieving both high sensitivity and a wide measuring range for concentration detection, as the measurable concentration range is restricted due to anisotropy saturation at high concentrations, and adjusting reagent amounts compromises sensitivity in the low-concentration region.
Innovation Solution
A measuring method involving sequential dispensing and measurement of fluorescent reagent, where the concentration of the measurement object is calculated based on anisotropy values using both initial and additional dispensing sequences, allowing for extended measuring range by adjusting reagent amounts to manage anisotropy saturation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the reagent amount is adjusted for high-sensitivity detection in the low-concentration region, then measurement sensitivity is improved, but the measurable concentration range is limited and anisotropy saturation occurs at high concentrations
Solution Approach 1:
The measurement process is divided into multiple sequences, each with different reagent dispensing amounts. The system segments the concentration measurement task into low-concentration detection (using higher reagent amounts for sensitivity) and high-concentration detection (using lower reagent amounts to avoid saturation), allowing both regions to be measured effectively
Solution Approach 2:
The reagent dispensing amount is made dynamic rather than fixed. The system automatically adjusts the reagent dispensing amount based on the measured anisotropy value and calibration data, selecting appropriate dispensing amounts from a set of predetermined values to optimize measurement for the current concentration range
2Adaptability or versatility
If the reagent amount is increased to extend the measurable concentration range to high concentrations, then the measuring range is expanded, but measurement sensitivity in the low-concentration region decreases
Solution Approach 1:
The system segments the concentration measurement task into multiple ranges, using different reagent dispensing amounts for different concentration regions. Multiple calibration curves are established corresponding to different dispensing amounts, allowing the system to select the appropriate calibration curve for the current measurement range
Solution Approach 2:
The reagent dispensing amount parameter is changed across multiple sequences to adapt to different concentration ranges. By varying this parameter, the system optimizes the measurement conditions for each specific concentration region, maintaining both sensitivity and range
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 measurement of concentration across a wide range, from low to high concentrations, by utilizing additional reagent dispensing to manage anisotropy values and maintain sensitivity, thereby overcoming the limitations of the initial method.
Implementation Method 1
The fluorescence polarization irradiates linearly polarized excitation light on a mixture (reaction solution) of a test sample containing an inspection item (object or antibody to be measured or a measurement object) and a fluorescent reagent, measures the fluorescent intensity emitted from the reaction solution through the polarization division
Implementation Method 2
measures the fluorescent intensity emitted from the reaction solution through the polarization division, and evaluates the polarization degree (polarization anisotropy or anisotropy). This anisotropy value is very sensitive to the rotational movement of the measurement object
Data Source
AI summary
A measuring method includes a first sequence for acquiring a first result about the anisotropy by performing a dispensing step and a measuring step and for measuring concentration of the measurement object from the first result based on a relationship between the anisotropy and a dispensing amount of the fluorescent reagent dispensed in the dispensing step, and a second sequence for acquiring a second result about the anisotropy by performing the dispensing step and the measuring step one or more times after the first sequence and for measuring the concentration of the measurement object from the second result based on the relationship between the anisotropy measured in the measuring step and the dispensing amount of the fluorescent reagent dispensed in the dispensing step.