Fluorescent Microscope Focus Scanning for Bright Point Detection
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Solution Overview
Problem
High numerical aperture in optical systems for fluorescent microscopes improves brightness and resolution but narrows focal depth, leading to blurred images of bright points distant from the focal plane, making accurate counting inefficient due to mixed brightness and size levels.
Innovation Solution
The focus position is moved in multiple preset scanning ranges with shorter lengths than the bright-point detection range, and images are taken and analyzed for high-frequency components to accurately count bright points in each cell nucleus.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the numerical aperture (NA) of the optical system is increased to improve brightness and resolution, then the brightness and resolution of bright points are improved, but the focal depth becomes narrow causing images of bright points distant from the focal plane to be blurred
Solution Approach 1:
The patent divides the detection process into multiple focal plane measurements. The image obtaining apparatus sequentially captures images at different focal planes (first focal plane, second focal plane, etc.) and processes them separately. This segmentation allows each measurement to be optimized for its specific focal depth while maintaining overall detection accuracy across the entire sample volume.
Solution Approach 2:
The patent transitions from two-dimensional image capture to three-dimensional volumetric detection by adding the focal depth dimension. By capturing images at multiple focal planes and synthesizing them, the system creates a comprehensive three-dimensional view of bright point distribution, enabling accurate counting regardless of the bright points' positions along the optical axis.
2Measurement precision
If images are taken at multiple focal planes to ensure all bright points are in focus, then detection accuracy is improved, but memory storage capacity and processing time are increased
Solution Approach 1:
The patent extracts only the essential information from multiple focal plane images. Instead of storing all raw image data, the system processes images at different focal planes to identify and count bright points, then stores only the extracted detection results. This extraction approach maintains high detection accuracy while significantly reducing memory storage requirements.
Solution Approach 2:
The patent discards redundant information from intermediate processing steps and recovers only the necessary detection data. By processing images sequentially and discarding temporary processing data while preserving essential bright point detection results, the system achieves accurate counting with minimal storage requirements.
3Productivity
If the focal depth is extended to capture all bright points in one image, then processing efficiency is improved, but resolution and brightness of individual bright points deteriorate
Solution Approach 1:
The patent segments the detection task into multiple focused measurements at different focal planes, with each measurement optimized for high resolution and brightness. By processing these segmented measurements sequentially and combining the results, the system achieves both high measurement precision and efficient overall detection without requiring extended focal depth.
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 enhances the accuracy of detecting fluorescent labels by obtaining clear images of bright points with uniform blurring, reducing the need for extensive memory storage and multiple data references, thus improving the efficiency of bright-point detection.
Implementation Method 1
a light source configured to irradiate a biological sample having a fluorescent label with an excitation light, the excitation light exciting the fluorescent label
Data Source
AI summary
An image obtaining apparatus includes: a light source configured to irradiate a biological sample having a fluorescent label with an excitation light, the excitation light exciting the fluorescent label; a focus moving unit configured to move a focus position of an optical system in the thickness direction of the biological sample; and a data processing unit configured to expose an image sensor to light while moving the focus position of the optical system in each of a plurality of preset scanning ranges to thereby obtain fluorescent images of the biological sample, each of the plurality of scanning ranges having a predetermined scanning length, the predetermined scanning length being smaller than the length of a bright-point detection range of the biological sample in the thickness direction, the center positions of the scanning ranges being different from each other in the thickness direction.