Immersion Objective Offset Calibration Without Immersion Liquid
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Solution Overview
Problem
Existing methods for ascertaining parfocality offset values in microscopy are complex, time-consuming, and prone to errors, especially when dealing with immersion objectives, which require the use of immersion liquids and result in increased calibration time and inaccuracies.
Innovation Solution
A method that allows for the determination of offset values for immersion objectives without the need for immersion liquids, using image representations from a z-stack to analyze contrast and resolution, and applying these values for parfocality and parcentricity corrections.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If immersion liquids are used to calibrate immersion objectives, then measurement precision is improved, but loss of time increases significantly
Solution Approach 1:
The patent extracts the immersion liquid from the calibration process entirely. Instead of requiring immersion liquid to be applied to the sample and objective, the method uses image representations taken without immersion liquid to determine offset values. This eliminates the time-consuming steps of applying and removing immersion liquid while maintaining calibration accuracy through alternative measurement approaches using z-stack analysis and contrast/resolution evaluation.
Solution Approach 2:
The patent creates a virtual model of the focal positions and offset values through image analysis. By analyzing image representations from z-stacks and evaluating contrast and resolution, the system generates offset values that replicate the information normally obtained through immersion-based calibration, thereby eliminating the need for physical immersion liquid application.
2Manufacturing precision
If manual correction methods are used to adjust objective positions, then manufacturing precision is improved, but ease of operation deteriorates
Solution Approach 1:
The system performs self-calibration by automatically determining offset values through image analysis. The microscope system itself, through its imaging capabilities and computational algorithms, identifies the focal positions and calculates the necessary correction values without requiring manual intervention or specialist knowledge from the user.
Solution Approach 2:
The patent replaces manual mechanical adjustment with an automated computational system. Instead of users physically adjusting objective positions or installing washers, the system uses image processing algorithms to analyze z-stacks, evaluate contrast and resolution, calculate offset values, and apply corrections automatically through the motorized focus drive.
3Measurement precision
If complex calibration procedures are implemented, then measurement precision is improved, but device complexity increases
Solution Approach 1:
The patent creates a universal calibration method that works for all objectives in the system, including both immersion and non-immersion objectives. The same image analysis workflow and offset determination algorithm can be applied across different objective types, eliminating the need for separate calibration procedures for each objective and simplifying the overall system complexity.
Solution Approach 2:
The system performs a complete calibration of all objectives using a standardized image analysis approach, rather than requiring selective or incremental calibration. By applying the same thorough analysis method to each objective, the system ensures precision without requiring users to navigate complex decision trees about which objectives need calibration or what specific procedures to follow for each case.
4Measurement precision
If immersion objectives are calibrated with immersion liquid, then measurement precision is improved, but productivity decreases due to sample removal and reapplication
Solution Approach 1:
The method extracts the requirement for immersion liquid from the calibration workflow. By determining offset values using image representations taken without immersion liquid, the system eliminates the steps of removing samples, applying immersion liquid, waiting for it to set, and then removing it again. This maintains focal position accuracy while allowing samples to remain in place throughout the calibration process.
Solution Approach 2:
The calibration process maintains continuous workflow without interruption. Samples remain mounted on the stage throughout the entire calibration procedure, and the imaging system continuously captures z-stack data for analysis. This eliminates the discontinuous steps of sample removal and reapplication, maintaining productive workflow while achieving accurate offset determination.
Data Source
AI summary
A method for ascertaining offset values for correcting the offset of an immersion objective of an image-creating optical system comprises the following steps: imaging an object using a reference objective of the optical system and the immersion objective without immersion and ascertaining offset values for an offset between the reference objective and the immersion objective with immersion on the basis of the image representations of the object