scholarly journals Quantitative 3D Reconstruction from Scanning Electron Microscope Images Based on Affine Camera Models

Sensors ◽  
2020 ◽  
Vol 20 (12) ◽  
pp. 3598 ◽  
Author(s):  
Stefan Töberg ◽  
Eduard Reithmeier
Keyword(s):  
Laser Scanning ◽  
Three Dimensional ◽  
Confocal Laser ◽  
Stereo Pair ◽  
Sem Images ◽  
Self Calibration ◽  
Dense Matching ◽  
Electron Microscopes ◽  
Affine Camera ◽  
Scanning Electron

Scanning electron microscopes (SEMs) are versatile imaging devices for the micro- and nanoscale that find application in various disciplines such as the characterization of biological, mineral or mechanical specimen. Even though the specimen’s two-dimensional (2D) properties are provided by the acquired images, detailed morphological characterizations require knowledge about the three-dimensional (3D) surface structure. To overcome this limitation, a reconstruction routine is presented that allows the quantitative depth reconstruction from SEM image sequences. Based on the SEM’s imaging properties that can be well described by an affine camera, the proposed algorithms rely on the use of affine epipolar geometry, self-calibration via factorization and triangulation from dense correspondences. To yield the highest robustness and accuracy, different sub-models of the affine camera are applied to the SEM images and the obtained results are directly compared to confocal laser scanning microscope (CLSM) measurements to identify the ideal parametrization and underlying algorithms. To solve the rectification problem for stereo-pair images of an affine camera so that dense matching algorithms can be applied, existing approaches are adapted and extended to further enhance the yielded results. The evaluations of this study allow to specify the applicability of the affine camera models to SEM images and what accuracies can be expected for reconstruction routines based on self-calibration and dense matching algorithms.

Testate Amoebae Examined by Confocal and Two-Photon Microscopy: Implications for Taxonomy and Ecophysiology

2010 ◽  
Vol 16 (6) ◽  
pp. 735-746 ◽  
Author(s):  
Zuzana Burdíková ◽  
Martin Čapek ◽  
Pavel Ostašov ◽  
Jiří Machač ◽  
Radek Pelc ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Three Dimensional ◽  
Environmental Scanning Electron Microscopy ◽  
Testate Amoebae ◽  
Shell Morphology ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Polarization Microscopy ◽  
Sem Images ◽  
Two Photon

AbstractTestate amoebae (TA) are a group of free-living protozoa, important in ecology and paleoecology. Testate amoebae taxonomy is mainly based on the morphological features of the shell, as examined by means of light microscopy or (environmental) scanning electron microscopy (SEM/ESEM). We explored the potential applications of confocal laser scanning microscopy (CLSM), two photon excitation microscopy (TPEM), phase contrast, differential interference contrast (DIC Nomarski), and polarization microscopy to visualize TA shells and inner structures of living cells, which is not possible by SEM or environmental SEM. Images captured by CLSM and TPEM were utilized to create three-dimensional (3D) visualizations and to evaluate biovolume inside the shell by stereological methods, to assess the function of TA in ecosystems. This approach broadens the understanding of TA cell and shell morphology, and inner structures including organelles and endosymbionts, with potential implications in taxonomy and ecophysiology.

3-Dimensional immunolabeling and in situ hybridization detection using fluorescence photooxidation and intermediate-voltage Electron Microscopy

1994 ◽  
Vol 52 ◽  
pp. 164-165
Author(s):  
Thomas J. Deerinck ◽  
Maryann E. Martone ◽  
Varda Lev-Ram ◽  
David P. L. Green ◽  
Roger Y. Tsien ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Reactive Oxygen ◽  
Confocal Laser Scanning Microscope ◽  
Fluorescent Dye ◽  
Confocal Laser ◽  
3 Dimensional ◽  
Voltage Electron ◽  
Dimensional Distribution ◽  
Electron Microscopes ◽  
New Generation

The confocal laser scanning microscope has become a powerful tool in the study of the 3-dimensional distribution of proteins and specific nucleic acid sequences in cells and tissues. This is also proving to be true for a new generation of high contrast intermediate voltage electron microscopes (IVEM). Until recently, the number of labeling techniques that could be employed to allow examination of the same sample with both confocal and IVEM was rather limited. One method that can be used to take full advantage of these two technologies is fluorescence photooxidation. Specimens are labeled by a fluorescent dye and viewed with confocal microscopy followed by fluorescence photooxidation of diaminobenzidine (DAB). In this technique, a fluorescent dye is used to photooxidize DAB into an osmiophilic reaction product that can be subsequently visualized with the electron microscope. The precise reaction mechanism by which the photooxidation occurs is not known but evidence suggests that the radiationless transfer of energy from the excited-state dye molecule undergoing the phenomenon of intersystem crossing leads to the formation of reactive oxygen species such as singlet oxygen. It is this reactive oxygen that is likely crucial in the photooxidation of DAB.

scholarly journals Investigation of Spin Coating Cerium-Doped Hydroxyapatite Thin Films with Antifungal Properties

Coatings ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 464
Author(s):  
Simona Liliana Iconaru ◽  
Mihai Valentin Predoi ◽  
Patrick Chapon ◽  
Sofia Gaiaschi ◽  
Krzysztof Rokosz ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Photoelectron Spectroscopy ◽  
Spin Coating ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
X Ray ◽  
Antifungal Properties ◽  
Scanning Electron

In this study, the cerium-doped hydroxyapatite (Ca10−xCex(PO4)6(OH)2 with xCe = 0.1, 10Ce-HAp) coatings obtained by the spin coating method were presented for the first time. The stability of the 10Ce-HAp suspension particles used in the preparation of coatings was evaluated by ultrasonic studies, transmission electron microscopy (TEM), X-ray diffraction (XRD), and scanning electron microscopy (SEM). The surface morphology of the 10Ce-HAp coating was studied by SEM and atomic force microscopy (AFM) techniques. The obtained 10Ce-HAp coatings were uniform and without cracks or unevenness. Glow discharge optical emission spectroscopy (GDOES) and X-ray photoelectron spectroscopy (XPS) were used for the investigation of fine chemical depth profiling. The antifungal properties of the HAp and 10Ce-HAp suspensions and coatings were assessed using Candida albicans ATCC 10231 (C. albicans) fungal strain. The quantitative antifungal assays demonstrated that both 10Ce-HAp suspensions and coatings exhibited strong antifungal properties and that they successfully inhibited the development and adherence of C. albicans fungal cells for all the tested time intervals. The scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) visualization of the C. albicans fungal cells adherence to the 10Ce-HAp surface also demonstrated their strong inhibitory effects. In addition, the qualitative assays also suggested that the 10Ce-HAp coatings successfully stopped the biofilm formation.

scholarly journals Three-dimensional relationships between tumor cells and microcirculation with double cyanine immunolabeling, laser scanning confocal microscopy, and computer-assisted reconstruction: an alternative to cast corrosion preparations.

1994 ◽  
Vol 42 (5) ◽  
pp. 681-686 ◽  
Author(s):  
V Rummelt ◽  
L M Gardner ◽  
R Folberg ◽  
S Beck ◽  
B Knosp ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Blood Vessels ◽  
Tumor Cells ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Melanoma Cells ◽  
Tumor Blood Vessels ◽  
The Relationship ◽  
Scanning Electron

The morphology of the microcirculation of uveal melanomas is a reliable market of tumor progression. Scanning electron microscopy of cast corrosion preparations can generate three-dimensional views of these vascular patterns, but this technique sacrifices the tumor parenchyma. Formalin-fixed wet tissue sections 100-150 microns thick from uveal melanomas were stained with the lectin Ulex europaeus agglutinin I (UEAI) and proliferating cell nuclear antigen (PCNA) to demonstrate simultaneously the tumor blood vessels and proliferating tumor cells. Indocarbocyanine (Cy3) was used as a fluorophore for UEAI and indodicarbocyanine (Cy5) was used for PCNA. Double labeled sections were examined with a laser scanning confocal microscope. Images of both stains were digitized at the same 5-microns intervals and each of the two images per interval was combined digitally to form one image. These combined images were visualized through voxel processing to study the relationship between melanoma cells expressing PCNA and various microcirculatory patterns. This technique produces images comparable to scanning electron microscopy of cast corrosion preparations while permitting simultaneous localization of melanoma cells expressing PCNA. The microcirculatory tree can be viewed from any perspective and the relationship between tumor cells and the tumor blood vessels can be studied concurrently in three dimensions. This technique is an alternative to cast corrosion preparations.

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