Application of Photogrammetric 3D Reconstruction to Scanning Electron Microscopy: Considerations for Volume Analysis

Photogrammetric three-dimensional (3D) reconstruction is an image processing technique used to develop digital 3D models from a series of two-dimensional images. This technique is commonly applied to optical photography though it can also be applied to microscopic imaging techniques such as scanning electron microscopy (SEM). The authors propose a method for the application of photogrammetry techniques to SEM micrographs in order to develop 3D models suitable for volumetric analysis. SEM operating parameters for image acquisition are explored and the relative effects discussed. This study considered a variety of microscopic samples, differing in size, geometry and composition, and found that optimal operating parameters vary with sample geometry. Evaluation of reconstructed 3D models suggests that the quality of the models strongly determines the accuracy of the volumetric measurements obtainable. In particular, they report on volumetric results achieved from a laser ablation pit and discuss considerations for data acquisition routines.

Download Full-text

A New Image Processing Technique for STEM

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100052171 ◽

1974 ◽

Vol 32 ◽

pp. 432-433

Author(s):

Yasushi Kokubo ◽

Hirotami Koike ◽

Teruo Someya

Keyword(s):

Electron Microscopy ◽

Image Processing ◽

Scanning Electron Microscopy ◽

Elastic Scattering ◽

Field Emission ◽

Processing Technique ◽

Image Processing Technique ◽

Energy Analyzer ◽

Scanning Microscope ◽

Scanning Electron

One of the advantages of scanning electron microscopy is the capability for processing the image contrast, i.e., the image processing technique. Crewe et al were the first to apply this technique to a field emission scanning microscope and show images of individual atoms. They obtained a contrast which depended exclusively on the atomic numbers of specimen elements (Zcontrast), by displaying the images treated with the intensity ratio of elastically scattered to inelastically scattered electrons. The elastic scattering electrons were extracted by a solid detector and inelastic scattering electrons by an energy analyzer. We noted, however, that there is a possibility of the same contrast being obtained only by using an annular-type solid detector consisting of multiple concentric detector elements.

Download Full-text

Scanning electron microscopy combined with image processing technique: Analysis of microstructure, texture and tenderness inSemitendinousandGluteus Mediusbovine muscles

Scanning ◽

10.1002/sca.21321 ◽

2016 ◽

Vol 38 (6) ◽

pp. 727-734

Author(s):

Facundo Pieniazek ◽

Valeria Messina

Keyword(s):

Electron Microscopy ◽

Image Processing ◽

Scanning Electron Microscopy ◽

Processing Technique ◽

Image Processing Technique ◽

Scanning Electron ◽

Technique Analysis

Download Full-text

Scanning electron microscopy combined with image processing technique: Microstructure and texture analysis of legumes and vegetables for instant meal

Microscopy Research and Technique ◽

10.1002/jemt.22626 ◽

2016 ◽

Vol 79 (4) ◽

pp. 267-275 ◽

Cited By ~ 2

Author(s):

Facundo Pieniazek ◽

Valeria Messina

Keyword(s):

Electron Microscopy ◽

Image Processing ◽

Scanning Electron Microscopy ◽

Texture Analysis ◽

Processing Technique ◽

Image Processing Technique ◽

Scanning Electron

Download Full-text

Scanning and Transmission Microscopy of Nematocyst Batteries in Epitheliomuscular Cells of Hydra

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100066097 ◽

1971 ◽

Vol 29 ◽

pp. 410-411 ◽

Cited By ~ 1

Author(s):

Jane A. Westfall ◽

S. Yamataka ◽

Paul D. Enos

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Osmium Tetroxide ◽

External Surface ◽

Three Dimensional ◽

Surface Structures ◽

Transmission Microscopy ◽

Transmission Electron ◽

Freeze Dried ◽

Scanning Electron

Scanning electron microscopy (SEM) provides three dimensional details of external surface structures and supplements ultrastructural information provided by transmission electron microscopy (TEM). Animals composed of watery jellylike tissues such as hydras and other coelenterates have not been considered suitable for SEM studies because of the difficulty in preserving such organisms in a normal state. This study demonstrates 1) the successful use of SEM on such tissue, and 2) the unique arrangement of batteries of nematocysts within large epitheliomuscular cells on tentacles of Hydra littoralis.Whole specimens of Hydra were prepared for SEM (Figs. 1 and 2) by the fix, freeze-dry, coat technique of Small and Màrszalek. The specimens were fixed in osmium tetroxide and mercuric chloride, freeze-dried in vacuo on a prechilled 1 Kg brass block, and coated with gold-palladium. Tissues for TEM (Figs. 3 and 4) were fixed in glutaraldehyde followed by osmium tetroxide. Scanning micrographs were taken on a Cambridge Stereoscan Mark II A microscope at 10 KV and transmission micrographs were taken on an RCA EMU 3G microscope (Fig. 3) or on a Hitachi HU 11B microscope (Fig. 4).

Download Full-text

Scanning Electron Microscopy and Electron Microprobe Analysis of Malignant Cell Cultures

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100100561 ◽

1968 ◽

Vol 26 ◽

pp. 164-165

Author(s):

R. I. Johnsson-Hegyeli ◽

A. F. Hegyeli ◽

D. K. Landstrom ◽

W. C. Lane

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Cell Cultures ◽

Electron Microprobe ◽

Microprobe Analysis ◽

Electron Microprobe Analysis ◽

Three Dimensional ◽

Malignant Cell ◽

Interference Microscopy ◽

Scanning Electron

Last year we reported on the use of reflected light interference microscopy (RLIM) for the direct color photography of the surfaces of living normal and malignant cell cultures without the use of replicas, fixatives, or stains. The surface topography of living cells was found to follow underlying cellular structures such as nuceloli, nuclear membranes, and cytoplasmic organelles, making possible the study of their three-dimensional relationships in time. The technique makes possible the direct examination of cells grown on opaque as well as transparent surfaces. The successful in situ electron microprobe analysis of the elemental composition and distribution within single tissue culture cells was also reported.This paper deals with the parallel and combined use of scanning electron microscopy (SEM) and the two previous techniques in a study of living and fixed cancer cells. All three studies can be carried out consecutively on the same experimental specimens without disturbing the cells or their structural relationships to each other and the surface on which they are grown. KB carcinoma cells were grown on glass coverslips in closed Leighto tubes as previously described. The cultures were photographed alive by means of RLIM, then fixed with a fixative modified from Sabatini, et al (1963).

Download Full-text

Faculty Opinions recommendation of Serial block-face scanning electron microscopy to reconstruct three-dimensional tissue nanostructure.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1022557.793507804 ◽

2015 ◽

Author(s):

Alpha Yap

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Three Dimensional ◽

Face Scanning ◽

Block Face ◽

Scanning Electron

Download Full-text

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.

Journal of Histochemistry & Cytochemistry ◽

10.1177/42.5.7908912 ◽

1994 ◽

Vol 42 (5) ◽

pp. 681-686 ◽

Cited By ~ 27

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.

Download Full-text