skip to main content
US FlagAn official website of the United States government
dot gov icon
Official websites use .gov
A .gov website belongs to an official government organization in the United States.
https lock icon
Secure .gov websites use HTTPS
A lock ( lock ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.


Title: Computer Vision Applied to In-Situ Specimen Orientation Adjustment for Quantitative SEM Analysis
Abstract Quantitative analysis methods based on the usage of a scanning electron microscope (SEM), such as energy-dispersive X-ray spectroscopy, often require specimens to have a flat surface oriented normal to the electron beam. In-situ procedures for putting microscopic flat surfaces into this orientation generally rely on stereoscopic methods that measure the change in surface vector projections when the surface is tilted by some known angle. Although these methods have been used in the past, there is no detailed statistical analysis of the uncertainties involved in such methods, which leaves an uncertainty in how precisely a specimen can be oriented. Here, we present a first principles derivation of a specimen orientation method and apply our method to a flat sample to demonstrate it. Unlike previous works, we develop a computer vision program using the scale-invariant feature transform to automate and expedite the process of making measurements on our SEM images, thus enabling a detailed statistical analysis of the method with a large sample size. We find that our specimen orientation method is able to orient flat surfaces with high precision and can further provide insight into errors involved in the standard SEM rotation and tilt operations.  more » « less
Award ID(s):
1900077
PAR ID:
10431417
Author(s) / Creator(s):
;
Date Published:
Journal Name:
Microscopy and Microanalysis
Volume:
29
Issue:
1
ISSN:
1431-9276
Page Range / eLocation ID:
196 to 201
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. Abstract Many attachments to a scanning electron microscope (SEM), such as energy dispersive x‐ray spectroscopy, extend its function significantly. Typically, the application of such attachments requires that the specimen has a planar surface at a specific orientation. It is a challenge to make the plane of a microscale specimen satisfy the orientation requirement since they are visible only in an SEM. An in‐situ procedure is needed to adjust specimen orientation by using stage rotation and tilting functions, in the process of which the key is to determine the initial orientation. This study proposed and tested top‐down and side‐view approaches to determine the orientation of a planar surface inside an SEM. In the top‐down one, the projected area is monitored on SEM images as stage rotation and tilt angles are adjusted. When the surface normal is along the electron beam direction, the area has a maximum value. In the side‐view approach, the stage is adjusted so that the projection appears to be a straight horizontal line on the SEM image. Once the orientation of the specimen for top‐down or side‐view observation is determined, the original can be calculated, and a desired orientation can be realized by manipulating the stage. The procedures have been tested by analyzing planar surfaces of spherical particles in Al‐Cu‐Fe alloy in the form of facets. The measured angles between two surfaces are consistent with those expected from crystallographic consideration within 2.7° and 1.7° for the top‐down and side‐view approaches, respectively. Research HighlightsTop‐down and side‐view approaches have been proposed and tested for in‐situ determination of specimen planar surface orientation in a Scanning Electron Microscope.The measured angles between two surfaces are consistent with those expected from crystallographic consideration within 2.7° and 1.7° for the top‐down and side‐view approaches, respectively. 
    more » « less
  2. Abstract. This paper introduces an innovative microscopy analysis methodology to preserve in situ non-soluble particles within hailstones using a protective porous plastic coating, overcoming previous limitations related to melting the hailstone sample. The method is composed of two techniques: trapping non-soluble particles beneath a plastic coat using the adapted sublimation technique and then analyzing the particles individually with confocal laser scanning microscopy (CLSM) and scanning electron microscopy with energy-dispersive spectroscopy (SEM–EDS). CLSM provides insights into physical attributes like particle size and surface topography, enhancing our understanding of ice nucleation. SEM–EDS complement CLSM by offering detailed information on individual particle elemental chemistry, enabling classification based on composition. Strategies to reduce background noise from glass substrates during EDS spectral analysis are proposed. By combining powerful, high-resolution microscopy techniques, this methodology provides valuable data on hailstone composition and properties. This information can give insights into hail developmental processes by enhancing our understanding of the role of atmospheric particles. 
    more » « less
  3. Crack formation limits the growth of (AlxGa1−x)2O3 epitaxial films on Ga2O3 substrates. We employ first-principles calculations to determine the brittle fracture toughness of such films for three growth orientations of the monoclinic structure: [100], [010], and [001]. Surface energies and elastic constants are computed for the end compounds—monoclinic Ga2O3 and Al2O3—and used to interpolate to (AlxGa1−x)2O3 alloys. The appropriate crack plane for each orientation is determined, and the corresponding critical thicknesses are calculated based on Griffith’s theory, which relies on the balance between elastic energy and surface energy. We obtain lower bounds for the critical thickness, which compare well with available experiments. We also perform an in-depth analysis of surface energies for both relaxed and unrelaxed surfaces, providing important insights into the factors that determine the relative stability of different surfaces. Our study provides physical insights into surface stability, crack planes, and the different degrees of crack formation in (AlxGa1−x)2O3 films for different growth orientations. 
    more » « less
  4. Abstract This study presents an experimental investigation to examine the mixed‐mode fracture behavior of fused filament fabrication printed acrylonitrile butadiene styrene (ABS). The single‐edge notch bending specimen configuration is employed to perform mixed‐mode fracture experiments. Four distinct printing orientations—90°, 0°, 45°/−45°, and 90°—are investigated. For each orientation, fracture studies are conducted under pure mode‐I loading (symmetric three‐point bending), mixed‐mode I/II, and pure mode‐II loading (asymmetric three‐point bending) to establish a mixed‐mode fracture criterion. The study evaluates the influence of printing orientation on fracture toughness, crack propagation behavior, and the mixed‐mode fracture criterion. Scanning electron microscopy (SEM) is utilized to analyze the fracture surfaces and correlate the observed fracture mechanisms with the measured fracture toughness values. The findings reveal that printing orientation significantly affects both the fracture toughness and the mixed‐mode fracture criterion. Among the orientations studied, the 90° specimens exhibit the highest fracture toughness and superior performance under all mixed‐mode conditions. SEM images of the fracture surfaces across different printing orientations show the formation of smooth shear zones of varying sizes near the crack tip under mixed‐mode and pure mode‐II conditions. These zones suggest an enhanced resistance to crack propagation, with the degree of improvement differing among the orientations. HighlightsMixed‐mode fracture behavior of 3D‐printed acrylonitrile butadiene styrene.Printing orientations have a major influence on mixed‐mode fracture criterion.90° printing orientation has the highest fracture toughness for mode‐mixities.0° printing orientation has the lowest fracture toughness for mode‐mixities.Fracture surface has dominant shear zone for all mode‐mixities except mode‐I. 
    more » « less
  5. Abstract Creating a high-resolution brain atlas in diverse species offers crucial insights into general principles underlying brain function and development. A volume electron microscopy approach to generate such neural maps has been gaining importance due to advances in imaging, data storage capabilities, and data analysis protocols. Sample preparation remains challenging and is a crucial step to accelerate the imaging and data processing pipeline. Here, we introduce several replicable methods for processing the brains of the gastropod mollusc,Berghia stephanieaefor volume electron microscopy. Although high-pressure freezing is the most reliable method, the depth of cryopreservation is a severe limitation for large tissue samples. We introduce a BROPA-based method using pyrogallol and methods to rapidly process samples that can save hours at the bench. This is the first report on sample preparation and imaging pipeline for volume electron microscopy in a gastropod mollusc, opening up the potential for connectomic analysis and comparisons with other phyla. 
    more » « less