Real-time PCR Monitoring Apparatus Using Polarization Separation

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Solution Overview

Problem

Conventional real-time PCR monitoring apparatuses face challenges in separating excitation light and fluorescence without using dichroic beam splitters, leading to interference from reflected excitation light, especially when using multiple fluorescent probes, as they require different dichroic beam splitters and struggle to completely separate excitation and fluorescence due to brighter excitation light.

Innovation Solution

The apparatus employs a polarization converter to convert excitation light into polarized light, using a polarizing beam splitter and polarizers to separate excitation and fluorescence based on polarization properties, eliminating the need for dichroic beam splitters and minimizing interference by ensuring excitation and fluorescence have different polarization components.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If a dichroic beam splitter is used to separate excitation light and fluorescence, then the separation is effective, but the system requires multiple different dichroic beam splitters for different fluorescent probes, increasing device complexity and cost

Engineering Contradiction:
Improveseparation of excitation light and fluorescenceVSAvoidnumber of dichroic beam splitters
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent changes the optical parameter of the excitation light from non-polarized to polarized state using a polarization converter. This parameter change enables the use of a single polarizing beam splitter to separate excitation light and fluorescence for multiple different fluorescent probes, eliminating the need for multiple dichroic beam splitters and resolving the contradiction between separation effectiveness and device complexity

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The polarizing beam splitter is designed to universally handle multiple fluorescent probes with different excitation wavelengths. By converting excitation light to polarized light first, a single polarizing beam splitter can perform the separation function for various probes, making the system multi-functional and reducing the number of components needed

Inventive Principle:
Principle #6Universality (Multi-functionality)

2Measurement precision

If excitation light intensity is increased to improve signal strength, then fluorescence detection sensitivity improves, but reflected excitation light interference increases

Engineering Contradiction:
Improvefluorescence detection sensitivityVSAvoidreflected excitation light interference
Core Design Contradiction:
Measurement precisionVSObject-generated harmful factors

Solution Approach 1:

The patent changes the polarization state of excitation light using a polarization converter and polarizers. This enables strong excitation light to be used for sensitive fluorescence detection while the polarizing beam splitter separates the polarized excitation light from fluorescence, preventing reflected excitation light interference from reaching the detector

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The polarizing beam splitter acts as an intermediary device that separates excitation light and fluorescence based on their different polarization states. This mediator allows high-intensity excitation light to illuminate the sample effectively while blocking reflected excitation light from reaching the detector, thus resolving the contradiction between signal strength and interference

Inventive Principle:
Principle #24Intermediary (Mediator)

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 effectively separates excitation light and fluorescence, reducing interference and allowing for real-time monitoring without the need for multiple dichroic beam splitters, improving the efficiency and economic viability of the system by using a single polarizing beam splitter for various fluorescent probes.

Implementation Method 1

a polarization converter (102) for converting an excitation light generated from a light source (100), into an excitation light with the polarization component

Methodology Applied
Scientific EffectPolarization: Polarisation

Implementation Method 2

a polarizing beam splitter (108) for separating the polarized light with a specific frequency passed through first polarizer (111)

Methodology Applied
Scientific EffectPolarizing beam splitting: Polarisation

Implementation Method 3

a first band pass filter (105) for allowing the polarized excitation light with a specific frequency of the surface light transferred from the light tunnel (104) that is matched with excitation properties of a fluorescent probe, to pass therethrough

Methodology Applied
Scientific EffectOptical filtering: Filter (optical)

Implementation Method 4

a second band pass filter (112) for allowing a fluorescence with a specific frequency of the polarized fluorescence transferred from the second polarizer (107) that is matched with an emitting property of a fluorescent probe, to pass therethrough

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Data Source

PatentEP2160476B1Real-time PCR monitoring apparatus
Publication Date: 2015.10.14 BIONEER
  • EP2160476B1 patent drawingFigure 1
  • EP2160476B1 patent drawingFigure 2~3
  • EP2160476B1 patent drawingFigure 4

AI summary

The present invention relates to a real-time PCR monitoring apparatus for real-time monitoring production of reaction product produced during the reaction while performing nucleic acid amplification such as PCR for various kinds of trace samples. Specifically, the present invention relates to an apparatus for real-time monitoring biochemical reaction for efficiently dividing interference between an excitation light and a fluorescence, which includes a polarizer, a polarizing beam splitter, a polarization converter and so on.