Image Analysis Using Focal Structures for Focus Accuracy
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Solution Overview
Problem
Current image-based analysis methods for detecting microscopic objects, such as those in flow cytometry, are complex and time-consuming, especially when dealing with large numbers of samples, as they require a separate focusing step for each image, which can significantly increase the time required for analysis.
Innovation Solution
A method utilizing a sample holder with multiple focal structures and a line camera that captures composite digital images of multiple locations of interest, allowing for rapid continuous focusing without checking focus for each frame, followed by image processing to ensure accurate focus and detect out-of-focus images, thereby reducing analysis time.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a separate focusing step is performed for each image in current image-based analysis methods, then focus accuracy is improved, but analysis time increases significantly
Solution Approach 1:
The patent applies preliminary action by capturing multiple frames with different focus settings in advance, before the actual analysis is needed. The system pre-captures a focus adjustment sequence that includes multiple frames at different focus positions, storing them for later retrieval. This allows the analysis to use pre-prepared focused images without performing time-consuming focusing operations at the moment of analysis, thus resolving the contradiction between focus accuracy and analysis time.
Solution Approach 2:
The patent uses copying by creating multiple copies of the sample at different focus positions within the same imaging session. Instead of repeatedly focusing on the same sample, the system captures multiple frames representing different focus states, effectively copying the sample's appearance at various focal planes. This eliminates the need for repeated focusing operations while preserving focus accuracy information across multiple captured copies.
2Productivity
If rapid continuous focusing is used without checking focus for each frame, then analysis speed is improved, but risk of out-of-focus images increases
Solution Approach 1:
The patent implements feedback by automatically analyzing captured frames to determine which ones are in focus. The system includes a focus determination module that evaluates each captured frame, identifies whether it is in focus or out of focus, and selects only the in-focus frames for analysis. This feedback mechanism ensures that even when using rapid continuous focusing without checking each frame, the system reliably identifies and uses only the properly focused images, thus maintaining focus quality while preserving analysis speed.
Solution Approach 2:
The system applies self-service by automatically performing focus determination and frame selection without requiring manual intervention. The focus determination module autonomously evaluates captured frames, identifies in-focus images, and proceeds with analysis using only those frames. This self-service capability allows the system to maintain high analysis speed while ensuring focus quality through automated quality control, resolving the contradiction between productivity and reliability.
3Productivity
If multiple samples are analyzed simultaneously using composite digital image areas, then throughput is improved, but complexity of image processing increases
Solution Approach 1:
The patent applies segmentation by dividing the composite digital image area into multiple discrete regions, each corresponding to a separate sample. The image processing system automatically segments the large composite image into individual sample regions, allowing each sample to be analyzed independently. This segmentation approach enables simultaneous analysis of multiple samples (improving throughput) while managing processing complexity by breaking down the overall task into smaller, manageable segments that can be processed in parallel or sequentially.
Data Source
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AI summary
A method of image-based analysis of multiple samples comprises: using a sample holder 100 having multiple locations of interest 16, 18 and multiple focal structures 12 that are each associated one or more with the multiple locations of interest 16, 18, wherein the multiple samples are dispersed across the multiple locations of interest 16, 18; obtaining image areas of the multiple locations of interest 16, 18, thereby obtaining multiple digital image areas for use in analysis of the multiple samples with each of the image areas including at least one of the locations of interest 16, 18 and at least one of the focal structures 12; using an image processing algorithm to analyse each of the digital image areas and check if the focal structure 12 indicates that the image area is in clear focus; and providing an indication and/or taking remedial action if the image processing algorithm indicates that any digital image areas are out of focus.