Microscopic Target Position Retrieval Using Multi-Resolution Reference Markers
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Solution Overview
Problem
Existing methods for focused ion beam scanning electron microscopy (FIB-SEM) in biological research face challenges in accurately retrieving target positions due to the limitations of using different resolution techniques, where position information from lower resolution methods like light microscopy is not usable to a sufficient extent in higher resolution methods like electron microscopy, and inaccuracies such as color errors and blurring occur.
Innovation Solution
A method involving providing a first digital representation of a sample at a first resolution, specifying target position identifiers, acquiring an image stack, generating a second digital representation at a higher resolution, and determining geometric descriptors between target and reference markers to enhance spatial resolution and improve position retrieval.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If lower-resolution light microscopy is used to identify target positions, then the overview and navigation are easier, but the measurement precision and spatial resolution are insufficient for high-resolution electron microscopy
Solution Approach 1:
The patent divides the digital representation into multiple resolution levels: a first digital representation at lower resolution for overview navigation, and a second digital representation at higher resolution for precise target identification. This segmentation allows users to navigate easily at low resolution then switch to high resolution only when needed, resolving the contradiction between ease of navigation and measurement precision.
Solution Approach 2:
The patent adds a resolution dimension to the digital representation system by creating multiple versions of the sample data at different resolutions. This dimensional approach allows the system to provide both easy navigation (using low-resolution data) and high measurement precision (using high-resolution data) simultaneously, rather than being constrained to a single resolution level.
2Measurement precision
If higher-resolution electron microscopy is used to achieve better spatial resolution, then the measurement precision improves, but the difficulty of detecting and measuring target positions increases
Solution Approach 1:
The patent performs preliminary identification of target positions using the first digital representation at lower resolution before acquiring the high-resolution image stack. This preliminary action allows the system to navigate to regions of interest and identify approximate target locations easily, then refine these positions with high-resolution data, reducing the difficulty of detecting and measuring targets in electron microscopy.
Solution Approach 2:
The first digital representation serves as an intermediary between the low-resolution overview and the high-resolution electron microscopy data. It mediates the transition by providing pre-identified target positions and regions of interest, making it easier to detect and measure targets in the high-resolution electron microscopy without having to search the entire sample space.
3Measurement precision
If multiple high-resolution image stacks are acquired to improve target position accuracy, then the measurement precision increases, but the productivity and data acquisition time increase
Solution Approach 1:
The patent applies high-resolution imaging only locally to specific regions of interest identified in the first digital representation, rather than acquiring high-resolution data for the entire sample. This local quality approach maintains high measurement precision for target positions while significantly reducing productivity loss by limiting high-resolution data acquisition to only the necessary areas.
Solution Approach 2:
The patent uses partial action by acquiring high-resolution image stacks only for selected regions of interest rather than the complete sample. This partial acquisition provides sufficient precision for target position identification without the excessive time cost of processing entire high-resolution datasets, resolving the contradiction between precision and productivity.
4Device complexity
If downscaling the second digital representation to match the first resolution is used to simplify processing, then the device complexity is reduced, but the loss of information occurs
Solution Approach 1:
The patent performs preliminary identification of target positions and reference markers in the high-resolution second digital representation before any downscaling occurs. This preliminary action ensures that all spatial detail information is captured and recorded at high resolution, so that when downscaling is performed for simplified processing, no critical information is lost because the high-resolution data has already been utilized for accurate position identification.
Data Source
AI summary
A method for providing position information for retrieving a target position in a microscopic sample includes providing a first representation of the sample at a first resolution including the target position; specifying a first target position identifier indicating the target position at the first resolution; acquiring an image stack comprising the target position indicated by the first target position identifier; providing a second representation at a second resolution higher than the first resolution based on the image stack; specifying a second target position identifier indicating the target position at the second resolution; specifying a plurality of reference position identifiers in the second representation indicating positions of optically detectable reference markers at the second resolution; and determining a set of geometric descriptors describing spatial relations between the second target position identifier and the plurality of reference position identifiers to provide the position information.