Fluorescence Image Processing for Luminescence Interference
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Solution Overview
Problem
Existing measuring apparatuses struggle to acquire low-noise fluorescence and luminescence images, especially when a luminescent substance with high luminescence levels is used, as the luminescence level can overwhelm the fluorescence signal.
Innovation Solution
A measuring apparatus and method that involves capturing a first image without excitation light to isolate luminescence and a second image with excitation light to capture fluorescence, then processing these images to remove the luminescence component, using an illumination optical system with a shutter and image processing to generate a low-noise fluorescence image.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a luminescent substance with high luminescence level is used, then the luminescence signal is enhanced, but the luminescence component overwhelms the fluorescence signal making it difficult to acquire low-noise fluorescence images
Solution Approach 1:
The patent segments the image acquisition process into three distinct steps: (1) acquiring a first image without excitation light to capture only luminescence, (2) acquiring a second image with excitation light to capture both fluorescence and luminescence, and (3) processing these images to separate the fluorescence component. This segmentation allows the harmful luminescence component to be isolated and removed, resolving the contradiction between enhanced luminescence signal and fluorescence image quality.
Solution Approach 2:
The patent extracts the luminescence component from the combined fluorescence and luminescence image by subtracting the first image (containing only luminescence) from the second image (containing both signals). This extraction process removes the harmful luminescence interference while preserving the desired fluorescence signal, enabling acquisition of low-noise fluorescence images even when high-luminescence substances are used.
2Measurement precision
If excitation light is continuously radiated to capture fluorescence, then fluorescence signal is acquired, but luminescence from the specimen cannot be separately measured
Solution Approach 1:
The patent employs periodic action by alternating the excitation light on and off during image acquisition. The excitation light is turned off during the first image acquisition to capture only luminescence, then turned on during the second image acquisition to capture both fluorescence and luminescence. This periodic switching enables separate measurement of luminescence and fluorescence signals, resolving the contradiction between fluorescence detection and luminescence signal separation.
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 the acquisition of low-noise fluorescence and luminescence images by effectively separating and removing the luminescence component from the fluorescence image, even when a luminescent substance with high luminescence levels is present.
Implementation Method 1
an illumination optical system that radiates excitation light coming from an excitation light source onto a specimen
Implementation Method 2
both fluorescence images and luminescence images can be acquired
Implementation Method 3
an image acquisition portion that acquires an image by measuring light generated at the specimen
Implementation Method 4
an image processing portion that, based on a first image, which is acquired by the image acquisition portion without radiating the excitation light, and a second image, which is acquired by the image acquisition portion while radiating the excitation light, generates a fluorescence image by removing a luminescence component from the second image
Data Source
AI summary
A low-noise fluorescence image and a low-noise luminescence image are acquired even if a luminescent substance having a high luminescence level is used. Provided is a measuring apparatus including an illumination optical system that radiates excitation light coming from an excitation light source onto a specimen A, an image acquisition portion that acquires an image by measuring light generated at the specimen, and an image processing portion that, based on a first image, which is acquired by the image acquisition portion without radiating the excitation light, and a second image, which is acquired by the image acquisition portion while radiating the excitation light, generates a fluorescence image by removing a luminescence component from the second image.