Distinct Optical Paths for Fluorescence Detection
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Solution Overview
Problem
Existing fluorescence detection systems use the same optical path for excitation and detection, resulting in reduced sensitivity due to beam splitter losses, limiting the ability to detect DNA amplification in qPCR processes efficiently.
Innovation Solution
A system with distinct optical paths for excitation and detection, where excitation light is directed through a central illumination tube and fluorescence is collected by optics surrounding the tube, eliminating the need for beam splitters and enhancing sensitivity by minimizing scattered light and losses.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Device complexity
If beam splitters are used to direct excitation light and collect fluorescence in the same optical path, then the system structure is simplified, but light loss increases and detection sensitivity decreases
Solution Approach 1:
The optical path is divided into two separate paths: an excitation light path and a fluorescence collection path. The excitation path delivers light through the sample well bottom, while the collection path gathers fluorescence from the sample, eliminating the need for beam splitters and reducing light loss in both paths.
Solution Approach 2:
The sample well bottom acts as an intermediary element that both transmits excitation light and allows fluorescence collection. This mediator enables the separation of excitation and detection paths while maintaining functional integration through the well structure.
2Adaptability or versatility
If beam splitters are used to separate excitation and detection paths, then the system can distinguish multiple fluorogenic probes, but only about one-quarter of the signal is measured due to transmission losses
Solution Approach 1:
The optical system is segmented into distinct excitation and collection paths, with the collection path optimized to capture maximum fluorescence signal. Multiple fluorogenic probes are distinguished through spectral filtering in the collection path rather than beam splitting, preserving signal intensity while maintaining multi-probe capability.
3Device complexity
If the same optical path is used for excitation and detection, then the system design is simpler, but scattered light from the excitation path interferes with fluorescence detection
Solution Approach 1:
The optical paths are segmented into separate excitation and collection channels. The excitation path is optimized for delivering light through the well bottom, while the collection path is positioned and filtered to capture only fluorescence, eliminating scattered light interference from the excitation path.
Solution Approach 2:
Scattered light from the excitation path is extracted and eliminated from the detection path. The collection path is designed to collect only fluorescence emitted at angles different from the excitation path, and optical filters are used to remove any remaining scattered excitation light.
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 increases the sensitivity and quality of fluorescence detection, allowing for earlier DNA detection after fewer thermal cycles, and is compatible with qPCR instruments, reducing the complexity and size of the detection system.
Implementation Method 1
a light source that emits an excitation light into an illumination tube
Implementation Method 2
fluorescence excitation and detection
Implementation Method 3
a plurality of collection optics located around an aperture in the illumination tube for collecting fluorescence
Implementation Method 4
a detector for determining the amount of fluorescence
Data Source
AI summary
A system and method for fluorescence excitation and detection having distinct optical paths is disclosed. A system for detecting fluorescence comprises a light source that emits an excitation light into an illumination tube; a plurality of collection optics located around an aperture in the illumination tube for collecting fluorescence; and a detector for determining the amount of fluorescence. A method for detecting fluorescence comprises emitting an excitation light from a light source into an illumination tube; directing the excitation light to an excitation filter; illuminating a sample with the excitation light to generate an emission light; and detecting the optical characteristics of the emission light using a plurality of collection optics located around the illumination tube.