Microscope Pattern Illumination Calibration for Coordinate Drift
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Solution Overview
Problem
Existing microscope-based systems face challenges in accurately illuminating patterns on samples due to long-term drift and misalignment between subsystems, leading to inaccuracies in pattern illumination and image processing, particularly in high-content applications requiring repeated processes across multiple regions.
Innovation Solution
An image-guided calibration method that adjusts the pattern illumination subsystem using correction factors derived from comparing actual and computed coordinates, ensuring accurate alignment and reducing the frequency of manual calibration by implementing a non-transitory computing device with instructions to adjust the position of light projection based on measured differences.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If manual calibration is performed frequently to maintain alignment accuracy, then pattern illumination precision is improved, but system downtime and operational complexity increase
Solution Approach 1:
The system performs automatic calibration using a calibration sample and image processing algorithms, eliminating the need for manual intervention. The calibration process is self-executing, where the system automatically detects pattern deviations and adjusts illumination coordinates without requiring operator involvement, thus resolving the contradiction between maintaining precision and minimizing downtime.
Solution Approach 2:
The system performs calibration automatically at predetermined intervals or before processing samples, proactively maintaining alignment accuracy before deviations affect actual sample analysis. This preliminary calibration action prevents precision degradation without requiring frequent manual corrections during operation.
2Measurement precision
If manual calibration is performed frequently to maintain subsystem alignment, then pattern illumination precision is improved, but operational efficiency deteriorates
Solution Approach 1:
The automatic calibration system operates autonomously without requiring operator intervention, performing calibration tasks in the background or during automated sequences. This eliminates the time operators would spend on manual calibration adjustments, thereby maintaining precision while preserving operational efficiency and sample processing throughput.
3Ease of operation
If subsystem alignment is not calibrated, then operational simplicity is maintained, but pattern illumination accuracy deteriorates
Solution Approach 1:
The system automatically performs calibration without requiring user intervention or knowledge of calibration procedures. Users simply operate the system normally, and the automatic calibration process handles alignment maintenance in the background, preserving operational simplicity while ensuring illumination accuracy through automated coordinate correction.
Solution Approach 2:
The system uses image processing to detect actual pattern positions, compares them with expected positions, and generates correction factors that are applied to subsequent illumination operations. This closed-loop feedback mechanism maintains accuracy automatically without requiring user awareness or intervention, balancing simplicity and precision.
4Loss of time
If automatic calibration is implemented, then calibration frequency is reduced, but system complexity increases
Solution Approach 1:
The system uses the existing imaging subsystem and processing capabilities to perform calibration functions, rather than adding dedicated calibration hardware. The same camera and image processing algorithms used for sample analysis are repurposed for calibration, achieving automatic calibration with minimal additional system complexity.
Solution Approach 2:
A calibration sample with known geometric patterns serves as an intermediary reference object that enables the system to self-calibrate. This calibration sample acts as a mediator between the illumination subsystem and imaging subsystem, allowing automatic coordinate alignment without complex direct measurement mechanisms.
Data Source
AI summary
A microscope based system for image-guided microscopic illumination is provided. The system may include a microscope, a stage, an imaging subsystem adapted to obtain an image of a sample on the stage, a processing subsystem adapted to identify regions of interest in the sample from images obtained by the imaging subsystem, and a pattern illumination subsystem adapted to illuminate the regions of interest based on coordinates derived from the images by the processing subsystem. Methods of calibrating the microscope based system may include projecting light from the pattern illumination subsystem onto the sample in the illumination pattern based on computed coordinates of the desired pattern, obtaining an image of the illumination pattern from the sample with the imaging subsystem, measuring differences between actual coordinates of the illumination pattern in the image and the computed coordinates, ‘and generating correction factors based on the measured differences to calibrate the system automatically.