Fluorescence Microscopy Focus Positioning via Axial Scanning
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Solution Overview
Problem
In fluorescence microscopy, accurately setting the focus position is challenging due to uneven sample thickness, slide glass thickness, and dust, especially with high-numerical-aperture optical systems, which complicates the imaging of biological samples with fluorescent labels, requiring inefficient manual adjustment and large memory storage for multiple image captures.
Innovation Solution
An image obtaining apparatus with a light source, optical system, image sensor, and data processing unit that moves the focus position within the imaging range to calculate distribution information of fluorescent labels, allowing efficient imaging by determining the optimal focus position based on fluorescent images captured during this movement.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the numerical aperture (NA) of the optical system is increased, then the brightness of bright points and the resolution of the image are improved, but the focal depth becomes narrow and the focus position becomes difficult to adjust
Solution Approach 1:
The system performs preliminary actions by capturing multiple images at different focus positions before final analysis. The focus position adjustment is automated through image analysis that identifies the optimal focus position from pre-captured images, eliminating the need for manual adjustment while maintaining high resolution
Solution Approach 2:
The system uses self-service by automatically determining the optimal focus position through image analysis. The captured images are processed to identify which image corresponds to the best focus position, allowing the system to self-adjust without manual intervention while maintaining high NA resolution
2Measurement precision
If the focus position is adjusted by capturing images at multiple intervals, then the accuracy of focus position detection is improved, but the memory capacity requirement increases and manual work time increases
Solution Approach 1:
The system changes the parameter of image capture by acquiring images at specific focus position intervals rather than continuously. This discrete parameter change reduces the total number of images needed while maintaining sufficient accuracy for focus position detection, thereby reducing memory requirements
Solution Approach 2:
The system extracts only the essential information needed for focus position determination from the captured images. By analyzing images at key intervals and extracting focus position data rather than storing and processing all captured images, the system reduces memory capacity requirements while maintaining detection accuracy
3Measurement precision
If the focus position is adjusted by capturing images at multiple intervals, then the accuracy of focus position detection is improved, but the time required for manual analysis increases
Solution Approach 1:
The system performs self-service by automatically analyzing captured images to determine the optimal focus position. The image analysis algorithm processes the images and identifies the best focus position without requiring manual review, thereby reducing manual analysis time while maintaining high detection accuracy
Solution Approach 2:
The system uses feedback from image analysis to automatically adjust and confirm the optimal focus position. The analysis of captured images provides feedback that guides the system to identify the correct focus position, eliminating the need for manual analysis while maintaining 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 method enables efficient imaging of biological samples with fluorescent labels by calculating the focus position automatically, reducing manual effort and memory requirements, while improving imaging accuracy and resolution.
Implementation Method 1
a fluorescent label, the fluorescent label emitting fluorescence when irradiated with excitation light
Data Source
AI summary
An image obtaining apparatus includes: a light source configured to irradiate a biological sample having a fluorescent label with excitation light, the excitation light exciting the fluorescent label; an optical system including an objective lens, the objective lens being configured to magnify an imaging target of the biological sample; an image sensor configured to form an image of the imaging target magnified by the objective lens; a movement controller configured to move a focus position of the optical system in an imaging range including at least a range corresponding to the thickness of the imaging target; and a data processing unit configured to exposure the image sensor to light while moving the focus position in the imaging range and obtain a fluorescent image of the biological sample, to thereby calculate distribution information of the fluorescent label in a thickness direction of the imaging target based on the fluorescent image.