Microscope Displacement Detection via Time-Division Fluorescence
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Solution Overview
Problem
Fiducial markers in super-resolution microscopy emit stronger fluorescence than the specimen, leading to reduced accuracy in detecting displacement and stage drift, which affects the precision of microscope observations.
Innovation Solution
A microscope device and control program that utilize a time division method to separately detect fluorescence from fiducial markers and the specimen, using distinct illumination wavelengths and intensities to minimize interference and enhance accuracy in displacement detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fiducial markers are used for displacement detection, then stage drift can be measured, but measurement precision deteriorates because fiducial markers emit stronger fluorescence than the specimen
Solution Approach 1:
The patent applies periodic action by alternately activating fiducial markers and specimen between different imaging periods. During the first period, fiducial markers are activated for displacement detection; during the second period, the specimen is activated for observation. This time-division multiplexing eliminates fluorescence interference while maintaining both displacement measurement and specimen observation capabilities.
Solution Approach 2:
The patent uses an intermediary approach by introducing a separate imaging period dedicated to fiducial marker detection. This intermediary period allows displacement measurement without the harmful fluorescence interference affecting the specimen observation period, effectively mediating between the need for accurate displacement detection and clean specimen imaging.
2Reliability
If activation light and excitation light conditions are optimized for specimen fluorescence, then specimen observation quality improves, but displacement detection accuracy deteriorates due to maximum detection value from fiducial markers
Solution Approach 1:
The patent implements periodic action by dividing imaging into distinct periods: one period optimized for fiducial marker detection with appropriate activation light conditions, and another period optimized for specimen observation with excitation light conditions. This periodic separation allows each parameter to be optimized independently without compromising the other.
Solution Approach 2:
The patent applies segmentation by separating the imaging process into distinct temporal segments or periods. Each segment is dedicated to a specific function (fiducial marker detection or specimen observation) with optimized lighting conditions, thereby eliminating the trade-off between specimen observation quality and displacement detection 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 allows for accurate detection of specimen behavior and stage displacement, improving the precision of microscope observations by reducing fluorescence interference from fiducial markers.
Implementation Method 1
The fluorescent substance becomes activate when irradiated with activation light and produces fluorescence or becomes inactivated when irradiated with excitation light in an activate state
Implementation Method 2
a detection value reaches maximum when a camera detects fluorescence emitted from fiducial markers
Data Source
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AI summary
[Problem] Provided is a microscope device that can accurately detect displacement of a microscope. [Solving means] The microscope device (1) includes an imager (6), an image processor (7), and a controller (42) . In a first period, the controller causes excitation light to be emitted and causes the imager to image a fluorescent image from an activated fluorescent substance in a plurality of frame periods. In a second period, the controller causes a fiducial marker to be irradiated with auxiliary light and causes the imager to image a fluorescent image from the fiducial marker, causes irradiation with the excitation light in the second period to stop or causes the intensity thereof to be reduced to be lower than that in the first period, and causes irradiation with the auxiliary light in the first period to stop or causes the intensity thereof to be reduced to be lower than that in the second period. The image processor uses an imaging result obtained in the second period to correct at least a part of an imaging result obtained in the first period and uses at least a part of the corrected imaging result to generate one image.