Fraunhofer in-line electron holography of frozen-hydrated DNA superhelices

1994 ◽  
Vol 52 ◽  
pp. 128-129
Author(s):  
Takao Matsumoto ◽  
Takayoshi Tanji ◽  
Akira Tonomura
Keyword(s):  
Electron Beam ◽  
High Resolution ◽  
Phase Shift ◽  
Heavy Atom ◽  
Specimen Preparation ◽  
Electron Holography ◽  
Small Object ◽  
Biological Specimen ◽  
Supercoiled Dna ◽  
Gold Particles

We have visualized DNA molecules in solution by Fraunhofer in-line electron holography. So far, applications of the technique at high resolution have been limited to gold particles. This report is the first application of Fraunhofer in-line electron holography to a nanometer-sized biological specimen.The phase shift of an electron beam due to DNA is so small, estimated as only 2π/33 for 100-keV electrons, that it has been only possible to visualize them with either heavy-atom staining or metal shadowing techniques. However, these methods give only low-resolution images of the molecules, and worst of all, preservation of specimens is poor. The frozen-hydrated specimen preparation improved the preservation of specimen, and images of naturally supercoiled DNA in solution have been visualized using underfocusing of 1-2 μm. Such a defocusing is large enough for such a small object as DNA (diameter ∽ 2 nm) to be in a Fresnel condition; z ≈ a2/λ, where z is the defocusing, a is the diameter of an object, and λ is the wavelength.

High-resolution observation of sintered alumina (Al2O3) using the specimen-heated and electron-beam-induced conductivity method in a UHR-SEM

1994 ◽  
Vol 52 ◽  
pp. 1046-1047
Author(s):  
K. Ogura ◽  
T. Suzuki ◽  
C. Nielsen
Keyword(s):  
Electron Beam ◽  
High Resolution ◽  
Specimen Preparation ◽  
Conductivity Method ◽  
Fine Grain ◽  
Grain Structures ◽  
Resolution Observation ◽  
Electron Microscopes ◽  
Scanning Electron Microscopes ◽  
Charging Effect

In spite of the complicated specimen preparation, Transmission Electron Microscopes (TEM) have traditionally been used for the investigation of the fine grain structures of sintered ceramics. Scanning Electron Microscopes (SEM) have not been used much for the same purpose as TEM because of poor results caused by the specimen charging effect, and also the lack of sufficient resolution. Here, we are presenting a successful result of high resolution imaging of sintered alumina (pure Al2O3) using the Specimen Heated and Electron Beam Induced Conductivity (SHEBIC) method, which we recently reported, in an ultrahigh resolution SEM (UHR-SEM). The JSM-6000F, equipped with a Field Emission Gun (FEG) and an in-lens specimen position, was used for this application.After sintered Al2O3 was sliced into a piece approximately 0.5 mm in thickness, one side was mechanically polished to get a shiny plane for the observation. When the observation was started at 20 kV, an enormous charging effect occured, and it was impossible to obtain a clear Secondary Electron (SE) image (Fig.1).

Electron Microscopic Observation of Biological Specimens in Their Native State by Employing Cryogenic Techniques

1972 ◽  
Vol 30 ◽  
pp. 410-411 ◽  
Author(s):  
Tokio Nei ◽  
Haruo Yotsumoto ◽  
Yoichi Hasegawa ◽  
Yuji Nagasawa
Keyword(s):  
Electron Microscopic Observation ◽  
Surface Structures ◽  
Specimen Preparation ◽  
Native State ◽  
Electron Microscopic ◽  
Biological Specimen ◽  
Specimen Holder ◽  
Cold Stage ◽  
Preparation Techniques ◽  
Scanning Electron

In order to observe biological specimens in their native state, that is, still containing their water content, various methods of specimen preparation have been used, the principal two of which are the chamber method and the freeze method.Using its recently developed cold stage for installation in the pre-evacuation chamber of a scanning electron microscope, we have succeeded in directly observing a biological specimen in its frozen state without the need for such conventional specimen preparation techniques as drying and metallic vacuum evaporation. (Echlin, too, has reported on the observation of surface structures using the same freeze method.)In the experiment referred to herein, a small sliced specimen was place in the specimen holder. After it was rapidly frozen by freon cooled with liquid nitrogen, it was inserted into the cold stage of the specimen chamber.

A Base Specific Single Heavy Atom Stain for Electron Microscopy

1972 ◽  
Vol 30 ◽  
pp. 184-185
Author(s):  
J. P. Langmore ◽  
N. R. Cozzarelli ◽  
A. V. Crewe
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
High Resolution ◽  
Elastic Scattering ◽  
Heavy Atom ◽  
High Vacuum ◽  
Heavy Atoms ◽  
Large Movement ◽  
Base Sequencing ◽  
Scanning Electron

A system has been developed to allow highly specific derivatization of the thymine bases of DNA with mercurial compounds wich should be visible in the high resolution scanning electron microscope. Three problems must be completely solved before this staining system will be useful for base sequencing by electron microscopy: 1) the staining must be shown to be highly specific for one base, 2) the stained DNA must remain intact in a high vacuum on a thin support film suitable for microscopy, 3) the arrangement of heavy atoms on the DNA must be determined by the elastic scattering of electrons in the microscope without loss or large movement of heavy atoms.

High Resolution Scanning Electron Microscopy

1991 ◽  
Vol 49 ◽  
pp. 478-479
Author(s):  
David Joy ◽  
James Pawley
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Electron Beam ◽  
Electron Microscope ◽  
High Resolution ◽  
Scanning Electron Microscope ◽  
Electron Probe ◽  
Impact Point ◽  
Interaction Volume ◽  
Scanning Electron

The scanning electron microscope (SEM) builds up an image by sampling contiguous sub-volumes near the surface of the specimen. A fine electron beam selectively excites each sub-volume and then the intensity of some resulting signal is measured. The spatial resolution of images made using such a process is limited by at least three factors. Two of these determine the size of the interaction volume: the size of the electron probe and the extent to which detectable signal is excited from locations remote from the beam impact point. A third limitation emerges from the fact that the probing beam is composed of a finite number of discrete particles and therefore that the accuracy with which any detectable signal can be measured is limited by Poisson statistics applied to this number (or to the number of events actually detected if this is smaller).

Quanitative aspects of electron diffraction using electron holography

1995 ◽  
Vol 53 ◽  
pp. 616-617
Author(s):  
E. Völkl ◽  
L.F. Allard ◽  
B. Frost ◽  
T.A. Nolan
Keyword(s):  
Electron Beam ◽  
Electron Diffraction ◽  
Software Package ◽  
Spatial Coherence ◽  
Electron Holography ◽  
Image Intensity ◽  
Interference Fringes ◽  
Complex Image ◽  
The Difference ◽  
Recorded Image

Off-axis electron holography has the well known ability to preserve the complex image wave within the final, recorded image. This final image described by I(x,y) = I(r) contains contributions from the image intensity of the elastically scattered electrons IeI (r) = |A(r) exp (iΦ(r)) |, the contributions from the inelastically scattered electrons IineI (r), and the complex image wave Ψ = A(r) exp(iΦ(r)) as:(1) I(r) = IeI (r) + Iinel (r) + μ A(r) cos(2π Δk r + Φ(r))where the constant μ describes the contrast of the interference fringes which are related to the spatial coherence of the electron beam, and Φk is the resulting vector of the difference of the wavefront vectors of the two overlaping beams. Using a software package like HoloWorks, the complex image wave Ψ can be extracted.

Ultrastructural analysis of the spatial distribution of MRNA

1992 ◽  
Vol 50 (1) ◽  
pp. 552-553
Author(s):  
Gary Bassell ◽  
Robert H. Singer
Keyword(s):  
Electron Microscopy ◽  
Spatial Distribution ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Nucleic Acids ◽  
Colloidal Gold ◽  
Dna Probe ◽  
Nucleotide Analogs ◽  
Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

Imaging of ferroelectric domain walls by electron holography

1992 ◽  
Vol 50 (1) ◽  
pp. 246-247
Author(s):  
Xiao Zhang
Keyword(s):  
Magnetic Field ◽  
High Resolution ◽  
Domain Walls ◽  
Ferroelectric Domain ◽  
Object Wave ◽  
Electron Holography ◽  
High Resolution Imaging ◽  
Quantitative Measurements ◽  
Resolution Imaging ◽  
Electron Microscopes

Electron holography has recently been available to modern electron microscopy labs with the development of field emission electron microscopes. The unique advantage of recording both amplitude and phase of the object wave makes electron holography a effective tool to study electron optical phase objects. The visibility of the phase shifts of the object wave makes it possible to directly image the distributions of an electric or a magnetic field at high resolution. This work presents preliminary results of first high resolution imaging of ferroelectric domain walls by electron holography in BaTiO3 and quantitative measurements of electrostatic field distribution across domain walls.

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