Fluorescent Polymer Calibration Plate for Plate Reader Deviation

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Solution Overview

Problem

There is a need for a method to calibrate fluorescence plate readers to reduce deviations in read-out values across different instruments, as existing methods do not effectively account for variations in light sources over time and between different plate readers.

Innovation Solution

A fluorescent polymer is cast on the wells of a plate, which is transparent, dry, and solvent- and porogen-free, using a specific dye and polymerization process that includes a concentration gradient, allowing for calibration through linear regression analysis to standardize read-out values.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional fluorescent dyes are used for calibration, then the calibration process can be performed, but the dyes are significantly quenched in aqueous solution or are unstable during illumination

Engineering Contradiction:

Improvestability of calibration standardVSAvoidquenching and instability of fluorescent dye

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent changes the physical and chemical parameters of the fluorescent material by embedding dyes within a polymer matrix. This structural modification protects the fluorescent molecules from quenching in aqueous solutions and stabilizes them during illumination, directly resolving the reliability issue with conventional dyes

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent creates a composite material system consisting of fluorescent dyes embedded in a polymer matrix. This composite structure combines the fluorescent properties of the dye with the protective and stabilizing characteristics of the polymer, eliminating the harmful effects of quenching and instability

Inventive Principle:

Principle #40Composite materials

2Adaptability or versatility

If different plate readers are used for measurements, then more measurement options are available, but there are significant deviations in read-out values due to different factory specifications and light source variations

Engineering Contradiction:

Improveavailability of different plate readersVSAvoiddeviation in read-out values

Core Design Contradiction:

Adaptability or versatilityVSMeasurement precision

Solution Approach 1:

The patent develops a universal calibration plate with a polymer-embedded fluorescent standard that can be used across different plate reader models. This single standardized reference material compensates for variations in light sources and detector specifications, enabling consistent measurements across multiple instruments

Inventive Principle:

Principle #6Universality (Multi-functionality)

Solution Approach 2:

The patent introduces a standardized fluorescent intensity parameter based on the polymer-embedded dye, which serves as a reference across different instruments. By changing from instrument-specific measurements to standardized reference-based measurements, the system achieves consistency across diverse plate readers

Inventive Principle:

Principle #35Parameter changes

3Ease of operation

If liquid standards are used for calibration, then the calibration can be performed, but the standards may evaporate or change concentration over time

Engineering Contradiction:

Improvesimplicity of calibration processVSAvoidstability of calibration standard over time

Core Design Contradiction:

Ease of operationVSDuration of action of stationary object

Solution Approach 1:

The patent transitions the calibration standard from liquid phase to solid polymer matrix phase. This phase change eliminates evaporation and concentration changes over time, while the solid polymer-embedded standard remains easy to handle and implement in the calibration process

Inventive Principle:

Principle #36Phase transitions

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

The method significantly reduces the coefficient of variation in read-out values among different plate readers, achieving consistent results with a coefficient of variation less than 5%, preferably less than 4%, and more preferably less than 2%, by providing a standardized calibration curve.

Implementation Method 1

Fluorescent polymer is a polymer that displays fluorescent properties... The fluorescent dye can be used in a variety of analytical methods, i.e., fluorescent imaging and spectroscopy

Methodology Applied

Scientific EffectFluorescence: Fluorescence

Implementation Method 2

an ethylenically unsaturated monomer is copolymerized with a polynuclear aromatic hydrocarbon or a substituted aromatic derivative thereof, such that the resultant polymer is fluorescent

Methodology Applied

Scientific EffectPolymerization: Photopolymerisation

Data Source

PatentUS12099011B2Method for calibrating fluorescent plate readers

Publication Date: 2024.09.24 PLEXBIO

AI summary

The present invention relates to a fluorescent polymer cast on the surface of wells of a plate and a plate comprising the fluorescent polymer. The plate is used in a method for calibrating read-out values of plate readers. The method for calibrating the plate readers can effectively reduce the deviation of read-out values among different plate readers.