Fluorescence Imaging Background Noise Reduction
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Solution Overview
Problem
Current fluorescence imaging systems face challenges in completely eliminating background noise, which affects the accuracy of target property measurements due to reflected and scattered excitation light, auto-fluorescence, and non-optical sources like dark signals, limiting their sensitivity and reliability.
Innovation Solution
The implementation of differential scanning methods with coaxial and angular illumination configurations, utilizing dichroic filters and array sensors to separate and subtract background noise from the target emission signal, allowing for accurate measurement of fluorescence signals by leveraging differences in optical properties and patterns between target and background light.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If optical filtering is used to separate emission light from excitation light, then the separation of emission and excitation light is improved, but reflected and scattered excitation light still reaches the sensor causing background noise
Solution Approach 1:
The patent extracts and removes the background noise component from the detected signal through differential scanning. By acquiring two images (one with excitation light and one without) and subtracting them, the system isolates and eliminates the background noise from reflected and scattered excitation light, leaving only the true emission signal.
Solution Approach 2:
The patent performs preliminary measurement of the background noise by acquiring an image without excitation light before or during the emission signal acquisition. This preliminary action allows the system to characterize and subsequently remove the background noise from the final measurement.
2Reliability
If multiple background noise sources are addressed with specific techniques, then the sensitivity for that specific background type is improved, but a general solution for all background noise types is lacking
Solution Approach 1:
The patent provides a universal solution that addresses all types of background noise (reflected excitation light, auto-fluorescence, dark signals) through a single differential scanning approach. The method works regardless of the noise source by simply comparing images with and without excitation light, making it adaptable to various fluorescence imaging configurations.
Solution Approach 2:
The patent converts the harmful background noise into a measurable quantity by capturing it as a separate image component. By treating the background noise as a distinct signal that can be measured and subtracted, the system transforms the problem from an unwanted interference into a manageable parameter that enhances overall measurement accuracy.
3Object-affected harmful factors
If hardware techniques are used to reduce background noise, then the root-cause of the problem is addressed, but the complexity of the system increases
Solution Approach 1:
The patent introduces a computational intermediary (software processing) that performs the background noise removal through image subtraction. Rather than adding complex hardware components, the system uses software as an intermediary to achieve background noise reduction, maintaining hardware simplicity while improving measurement accuracy.
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 reduces or eliminates background noise, enhancing the sensitivity and accuracy of fluorescence imaging by isolating the target signal from background interference, resulting in improved measurement precision and reduced noise levels.
Implementation Method 1
the target to be imaged is illuminated by an optical signal having a first spectral content (excitation light), and a portion of such a signal is absorbed by at least part of the target and is re-emitted as an optical signal of a second spectral content (emission light)
Implementation Method 2
One common method for separating the emission light from reflected and/or scattered excitation light is optical filtering
Data Source
Figure 1A~2B
Figure 3A~4B
Figure 5
AI summary
Systems and methods for producing background-reduced fluorescence imaging signals. An illumination system provides illumination light from an illumination source to a targeted area on the sample platform. Collection optics are configured to project light emanating from the sample platform onto the sensor adapted to detect light and having an array of sensing locations. In typical operation, light from the targeted area is projected onto a first portion of the sensor including a first plurality of the sensing locations and light from proximal to the targeted area on the platform is projected onto a second portion of the sensor including a second plurality of the sensing locations. A second signal detected by the second portion of the sensor is subtracted from a first signal detected by the first portion of the sensor to produce a background-reduced signal, e.g., a signal with reduced background related noise.