Sample Preparation With Multi-Angle ROI Localization
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Solution Overview
Problem
Accurately localizing a region of interest (ROI) in a 3D bulk sample for preparation under a charged particle microscope is challenging due to the limited axial resolution of fluorescence imaging systems, which hinders efficient sample preparation for transmission electron microscopy (TEM).
Innovation Solution
Acquiring sample images from different light incidence angles to determine the ROI location with higher spatial resolution, allowing for precise milling and micromachining of the sample while maintaining the same coordinate system for imaging and milling, thereby enhancing axial localization accuracy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescence imaging is used to localize ROI in 3D bulk sample, then sample preparation can be performed, but the axial resolution is limited which reduces localization accuracy
Solution Approach 1:
The patent acquires optical images from multiple viewing angles (different axial positions) to reconstruct 3D information. By capturing images at multiple focal depths and combining them, the system overcomes the limited axial resolution of single-plane fluorescence imaging, enabling accurate 3D localization of the ROI without requiring higher axial resolution in a single imaging plane.
2Measurement precision
If multiple imaging methods are used to improve ROI localization accuracy, then measurement precision improves, but the process becomes more complex and time-consuming
Solution Approach 1:
The patent employs a single optical microscope system that performs multiple functions: acquiring images at different focal depths, capturing images from different viewing angles, and providing 3D localization information. This multi-functional approach eliminates the need for multiple specialized imaging systems, reducing overall system complexity while maintaining high localization accuracy.
Solution Approach 2:
The patent combines multiple 2D optical images acquired at different focal depths and viewing angles into a unified 3D localization of the ROI. By merging information from multiple images taken with the same microscope system, the method achieves accurate 3D localization without requiring complex multi-system setups.
3Productivity
If traditional fluorescence imaging is used, then sample preparation can proceed, but the axial resolution limitation hinders efficient and accurate sample preparation for TEM
Solution Approach 1:
The patent performs preliminary 3D localization of the ROI using optical imaging from multiple angles and focal depths before initiating the milling process. This preliminary action ensures that the region of interest is accurately identified and localized in three dimensions, allowing subsequent milling operations to be precisely targeted, thereby improving overall sample preparation efficiency and accuracy.
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 enables accurate and quick localization and preparation of the sample, allowing for higher spatial resolution in ROI mapping and milling, ensuring that the ROI is accurately exposed for further analysis under a charged particle microscope without breaking the vacuum.
Implementation Method 1
irradiating a sample held by a sample stage along a first axis relative to the sample with a light beam and acquiring at least a first sample image by detecting emitted photons from the sample
Implementation Method 2
milling the sample held by the sample stage based on the location of the ROI
Implementation Method 3
milling the sample held by the sample stage based on the location of the ROI
Data Source
AI summary
A sample is milled to expose the region of interest (ROI) within the sample, while being held by a sample stage in a microscopy system. The sample is milled based on the ROI location determined with sample images acquired with light beam irradiating from different axes. The sample images are acquired while the sample is held using the same sample stage for milling.