Microscope Optical Alignment Using Multi-Region Contrast Focus
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Solution Overview
Problem
Existing microscope systems often fail to maintain high image quality across the entire field of view due to tilting between the sample plane and the focal plane, especially with large numerical aperture objectives, and this degradation is exacerbated by shipping, handling, and improper sample loading.
Innovation Solution
A method for image-based detection of optical alignment that involves dividing the field of view into sub-regions, determining contrast distribution for each sub-region, and generating angular discrepancy based on in-focus positions to adjust optical alignment and improve focus across the entire field of view.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If a single figure of merit is derived from the entire subject or a portion of the field of view for auto-focusing, then the focusing process is simplified and faster, but image quality degradation occurs at edges or corners of the field of view due to tilting between the sample plane and the focal plane
Solution Approach 1:
The field of view is divided into multiple sub-regions (e.g., central region and peripheral regions), and a separate figure of merit is calculated for each sub-region. This allows independent optimization of focus for different areas, enabling the system to detect and correct angular discrepancies that cause edge degradation while maintaining efficient automated focusing.
2Measurement precision
If direct methods for auto-focusing measure image contrast and high-frequency components, then the focusing accuracy is improved, but the system becomes more sensitive to angular discrepancies and tilting between sample plane and focal plane
Solution Approach 1:
By segmenting the field of view into multiple sub-regions and calculating figures of merit for each, the system maintains high focusing accuracy through direct measurement methods while becoming robust to angular discrepancies. The multi-region approach allows detection of tilting patterns, enabling the system to compensate for alignment disturbances rather than being sensitive to them.
3Reliability
If indirect methods for auto-focusing use hardware configurations such as high-precision motorized mechanical stage with position encoder, then the focusing process is more stable, but the device complexity increases
Solution Approach 1:
The patent replaces complex mechanical positioning systems with an image-based computational approach. Instead of relying on high-precision motorized stages and position encoders, the system uses contrast analysis of image data from multiple sub-regions to determine focus quality and detect angular discrepancies, achieving stable focusing with simpler hardware.
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 enhances imaging quality and throughput by accurately correcting angular discrepancies between the sample and focal planes, ensuring sharp focus throughout the entire field of view.
Implementation Method 1
determining a contrast distribution for a plurality of divided sub-regions for the field of view based on the plurality of contrast samples; and, generating an amount of angular discrepancy based on an in-focus position derived from the contrast distribution
Data Source
AI summary
Methods and systems are provided for a microscope system. In one example, a method for image-based detection of optical alignment for the microscope system comprises acquiring a plurality of contrast samples of a field of view of a subject, determining a contrast distribution for a plurality of divided sub-regions for the field of view based on the plurality of contrast samples, and generating an amount of angular discrepancy based on an in-focus position derived from the contrast distribution for each of the plurality of divided sub-regions. In one example, the method further comprises adjusting the microscope system based on the amount of angular discrepancy.