Microscope Shading Correction Using Peripheral Image Stitching
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Solution Overview
Problem
Existing microscope systems face challenges with cumbersome calibration processes, requiring temporary removal of samples or dedicated calibration samples, which can be damaged or contaminated, and struggle with generating uniform fluorescence samples for effective shading correction.
Innovation Solution
A microscope system that captures a reference viewing field image and multiple peripheral viewing field images by moving the sample relative to the optical system, calculates correction gains for each pixel based on these images, and applies shading correction using a correction gain calculation section and shading correction section, eliminating the need for sample removal or dedicated calibration samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a sample is temporarily removed to obtain calibration image data, then shading correction can be performed, but the operation becomes cumbersome and time-consuming
Solution Approach 1:
The system uses the sample itself to generate calibration image data by capturing peripheral viewing field images that contain the sample's own fluorescent signal. This eliminates the need to remove the sample for calibration, as the sample serves its own calibration purpose through its inherent fluorescence properties.
Solution Approach 2:
The system performs preliminary capture of peripheral viewing field images that include the sample's fluorescent signal before final image stitching. These preliminary images are used to calculate correction gains that compensate for shading, ensuring correction is built into the image processing pipeline before final composition.
2Measurement precision
If a dedicated calibration sample is used, then shading correction can be performed, but sample management becomes cumbersome due to damage and dust contamination risks
Solution Approach 1:
The system eliminates dedicated calibration samples by using the actual sample's fluorescent signal to generate calibration data. The sample serves itself for calibration purposes, eliminating the need for separate calibration artifacts that are susceptible to damage and contamination.
Solution Approach 2:
The system creates a virtual calibration reference by capturing peripheral images of the actual sample and using its fluorescent signal characteristics as the basis for correction gains, rather than relying on physical calibration samples that can be damaged or contaminated.
3Measurement precision
If a uniform fluorescence sample is generated for calibration, then accurate shading correction can be achieved, but generating such a sample is difficult and time-consuming
Solution Approach 1:
The system uses the actual sample's fluorescent signal as captured in peripheral viewing field images to generate correction gains, eliminating the need to manually create or verify uniform fluorescence calibration samples. The sample's own signal characteristics are directly utilized for correction purposes.
Solution Approach 2:
The system performs preliminary capture of peripheral images containing the sample's fluorescent signal and automatically processes these to generate correction gains, eliminating the manual time-consuming process of creating and verifying uniform fluorescence calibration samples while maintaining correction accuracy.
Data Source
AI summary
A microscope system comprises: a stage carrying a sample; an optical system forming an sample image; a driver driving at least the optical system or stage to relatively moves the sample and optical system; an imaging section capturing a reference viewing field image as an image of a predetermined viewing field range of the sample and peripheral viewing field images each being an image of a peripheral viewing field range containing a predetermined region in the predetermined viewing field range and different from the predetermined viewing field range, by the driver moving the relative position of the sample; a correction gain calculator calculating a correction gain of each pixel of the reference viewing field image based on the reference viewing field image and peripheral viewing field image; and a corrector performing shading correction on the reference viewing field image based on the calculated correction gain.