Fluorescence Microscope Imaging Modes for Cross-Excitation Control
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Solution Overview
Problem
Existing fluorescence microscope systems face challenges in imaging multiple fluorophores simultaneously due to cross-excitation, bleaching, and non-linear effects, which degrade image quality and require time-consuming filter and light source changes.
Innovation Solution
A fluorescence microscope system with a control unit that determines the optimal imaging mode (concurrent or sequential) based on fluorophore characteristics and system parameters, allowing simultaneous or subsequent imaging of fluorophores to minimize cross-excitation and enhance image quality through spectral unmixing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If multiple fluorophores are imaged simultaneously using multiple excitation light sources and detectors, then imaging speed is improved, but cross-excitation among fluorophores and non-linear effects like bleaching occur which degrade image quality
Solution Approach 1:
The patent segments the imaging process by dividing multiple fluorophores into different groups that are imaged sequentially rather than simultaneously. The control unit manages separate acquisition cycles for different fluorophore groups, eliminating cross-excitation while maintaining efficient imaging throughput through automated switching between groups.
Solution Approach 2:
The patent implements periodic action by using alternating illumination cycles where different excitation light sources are activated in alternating periods. The control unit switches between excitation sources and detectors in a periodic manner, allowing each fluorophore group to be excited only during its designated time window, thus preventing cross-excitation while maintaining high imaging speed.
2Reliability
If filter and light source changes are made to image different fluorophores individually, then image quality is improved, but a lot of time passes between acquisition of different images
Solution Approach 1:
The patent applies preliminary action by pre-configuring multiple excitation light sources and detectors to match different fluorophore groups before imaging begins. The control unit has already prepared the appropriate illumination and detection settings for each fluorophore group, so during acquisition it only needs to switch between pre-configured channels rather than adjusting filters and light sources in real-time, significantly reducing acquisition time while maintaining optimal image quality for each fluorophore.
3Productivity
If high intensity illumination is used to excite multiple fluorophores simultaneously, then imaging speed is improved, but non-linear effects like bleaching occur which degrade image quality
Solution Approach 1:
The patent segments the illumination intensity application by directing high intensity light only to the specific fluorophore group being imaged during each time window, rather than illuminating all fluorophores simultaneously. The control unit activates only the relevant excitation light source for the current fluorophore group, maintaining high imaging speed through focused illumination while preventing bleaching by avoiding unnecessary exposure of other fluorophores.
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 system ensures high-quality imaging by preventing cross-excitation and bleaching, enabling efficient acquisition of images without requiring specialized user knowledge.
Implementation Method 1
an optical detection system configured to generate images of the sample based on fluorescence light emitted by the excited at least two different fluorophores
Data Source
AI summary
A fluorescence microscope system for imaging a sample having at least two different fluorophores includes an illumination system configured to emit illumination light for exciting the fluorophores; an optical detection system configured to generate images of the sample based on fluorescence light emitted by the excited fluorophores; and a control unit configured to determine whether to image the sample in a concurrent imaging mode or in a sequential imaging mode, based on at least one characteristic of each of the fluorophores and based on at least one parameter of the optical detection system and/or the illumination system. In the concurrent imaging mode, the fluorophores are imaged simultaneously, and in the sequential imaging mode, the fluorophores are divided into a first group and at least one second group, and fluorophores of the first group and the at least one second group are imaged subsequently.