Laser Microdissection Calibration Using Digital Image Feedback
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Solution Overview
Problem
Existing laser microdissection systems face challenges in accurately calibrating laser deflection devices, leading to inconsistencies between target and actual laser positions, which affects the precision and cleanliness of sample extraction.
Innovation Solution
A method for calibrating a laser deflection device in a laser microdissection system using a digital image capturing unit and image evaluation module, where actuation signals are calculated based on default position values and calibration marks are made on a calibration object to determine actual position values, allowing for the calculation of second calibration values that align target and actual positions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If a laser deflection device is used to direct the laser beam over a stationary object, then the object remains stationary and stable during cutting, but the accuracy of laser positioning may deviate from default position values due to calibration errors
Solution Approach 1:
The patent applies preliminary action by performing calibration before actual laser microdissection operations. A calibration object with known geometric features is positioned in the object plane, and the laser deflection device is calibrated using these features to establish accurate correspondence between default position values and actual laser impact positions. This preliminary calibration ensures that subsequent cutting operations achieve high positioning accuracy while the object remains stationary.
2Device complexity
If manual calibration methods are used for the laser deflection device, then the system structure remains simple, but the calibration accuracy and reliability are insufficient
Solution Approach 1:
The patent introduces a calibration object as an intermediary element between the laser deflection device and the control system. This calibration object contains precisely manufactured geometric features (such as patterns, markers, or structures with known dimensions) that serve as reference standards. By using this intermediary, the system can automatically determine the relationship between default position values and actual laser impact positions, significantly improving calibration accuracy without requiring complex manual adjustment procedures.
3Ease of operation
If calibration marks are made manually, then the process is simple, but time is lost and precision is reduced
Solution Approach 1:
The patent replaces manual mechanical calibration operations with an automated optical-digital system. Instead of manually positioning and marking calibration features, the system uses a digital image capturing device to automatically detect the calibration object's geometric features, and software algorithms to calculate the transformation between default and actual positions. This substitution of manual mechanical operations with automated optical-digital processes dramatically reduces calibration time while maintaining ease of operation through software-driven procedures.
4Stability of the object's composition
If the laser beam is moved by moving the microscope stage, then the laser beam remains stationary, but the object moves in the field of vision causing blurred images
Solution Approach 1:
The patent applies inversion by reversing the conventional approach: instead of moving the microscope stage to follow the laser beam, the laser beam is deflected across the stationary object using a laser deflection device. This inversion maintains the object's stationary position in the field of vision, ensuring continuous clear imaging during the cutting process, while the laser beam dynamically positions itself to follow the desired cutting path.
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 method enhances the accuracy and reliability of laser calibration, reducing errors and improving the precision of sample extraction by automatically determining and adjusting laser positions, thereby increasing the cleanliness and precision of sample collection.
Implementation Method 1
making at least one calibration mark on the calibration object using the laser beam
Implementation Method 2
capturing an image of the calibration object by the digital image capturing unit
Data Source
AI summary
A method for calibrating a laser deflection device in a reflected light device of a microscope of a laser microdissection system having a digital image capturing unit comprising an image evaluation module includes generating a laser beam; guiding the laser beam through a microscope objective; directing the laser beam to a position defined by actuation signals; placing a calibration object in the object plane of the microscope objective; actuating the laser deflection device using first actuation signals and first calibration values, making at least one calibration mark on the calibration object; capturing an image of the calibration object by the digital image capturing unit; determining actual position values for the at least one calibration mark: and determining second calibration values based on a relationship between the default position values and the actual position values.