Comparison of Conventional and Electron Spectroscopic Imaging in the Fixed Beam Electron Microscope of Ordered Protein Arrays

Author(s):  
E. L. Buhle ◽  
U. Aebi

CTEM brightfield images are formed by a combination of relatively high resolution elastically scattered electrons and unscattered and inelastically scattered electrons. In the case of electron spectroscopic images (ESI), the inelastically scattered electrons cause a loss of both contrast and spatial resolution in the image. In the case of ESI imaging on the Zeiss EM902, the transmited electrons are dispersed into their various energy components by passing them through a magnetic prism spectrometer; a slit is then placed in the image plane of the prism to select the electrons of a given energy loss for image formation. The purpose of this study was to compare CTEM with ESI images recorded on a Zeiss EM902 of ordered protein arrays. Digital image processing was employed to analyze the average unit cell morphologies of the two types of images.

Author(s):  
J. M. Cowley

The resolution attained in an electron microscope image may be judged in a number of ways, depending on the means of interpretation used. The capabilities of available instruments are now approaching the point where a clear distinction between types of limitation of resolution and interpretive processes becomes important. Until recently the attainable resolution was limited by incoherent blurring of the image intensity by chromatic aberration effects due to electron energy spread and by electrical and mechanical instabilities of the microscope. Then the incoherent “spread function” defines the resolution, which may possibly be improved by ‘image deblurring’ by optical or computerized image processing. Now with the best microscopes the effects of the incoherent blurring can be reduced until the dominating effect in the limitation on resolution comes from coherent perturbations of the image due to defocus and spherical and other aberrations.


Author(s):  
Wah Chiu ◽  
Robert M. Glaeser

One of the objectives of our research program is to obtain a 2.0 Å point to point resolution in a fixed beam bright field electron microscope. The resolution in the fixed beam electron microscope is limited by a number of factors: electron beam coherence, energy spread, objective lens stability, mechanical stability, and specimen stability. This paper presents systematic studies of the mentioned factors in our JEM 100B fixed beam electron microscope equipped with a field emission gun operating at ∼ 1800°K.The most important characteristic of a field emission gun is its high brightness in the emitter source. In order to estimate the brightness at the specimen plane, one needs to measure the electron beam current density and the angle of illumination. The electron beam current density has been measured by means of a lithium-drifted silicon detector located below the normal position of the photographic plates. The angle of illumination can be estimated from the size of the condenser aperture and its distance from the specimen plane.


Author(s):  
D.P. Bazett-Jones ◽  
M.L. Brown

Elemental distributions in cells and molecular spreads can now be produced at the spatial resolution attainable in the electron microscope by the collection of X-rays or by the collection of and imaging with inellastically scattered electrons. With the latter method, known as Electron Spectroscopic Imaging (ESI), an image is produced with electrons that have lost characteristic amounts of energy from ionizing or exciting specific elements in the specimen. ESI can generate an elemental map of a specimen at a resolution of about 0.5 nm. It can be carried out in a fixed beam microscope equipped with a parallel energy filter inserted into the column of the microscope below the specimen (1,2). An instrument equipped with a prism-mirror-prism electron spectrometer was used in this study to image purified DNA molecules and a complex of the transcription factor TFIIIA with DNA.Transcription of most genes is activated by the binding of transcription factors to promoter elements.


Author(s):  
A. V. Crewe

The high resolution STEM is now a fact of life. I think that we have, in the last few years, demonstrated that this instrument is capable of the same resolving power as a CEM but is sufficiently different in its imaging characteristics to offer some real advantages.It seems possible to prove in a quite general way that only a field emission source can give adequate intensity for the highest resolution^ and at the moment this means operating at ultra high vacuum levels. Our experience, however, is that neither the source nor the vacuum are difficult to manage and indeed are simpler than many other systems and substantially trouble-free.


Author(s):  
Louis T. Germinario

A liquid nitrogen stage has been developed for the JEOL JEM-100B electron microscope equipped with a scanning attachment. The design is a modification of the standard JEM-100B SEM specimen holder with specimen cooling to any temperatures In the range ~ 55°K to room temperature. Since the specimen plane is maintained at the ‘high resolution’ focal position of the objective lens and ‘bumping’ and thermal drift la minimized by supercooling the liquid nitrogen, the high resolution capability of the microscope is maintained (Fig.4).


Author(s):  
Mihir Parikh

It is well known that the resolution of bio-molecules in a high resolution electron microscope depends not just on the physical resolving power of the instrument, but also on the stability of these molecules under the electron beam. Experimentally, the damage to the bio-molecules is commo ly monitored by the decrease in the intensity of the diffraction pattern, or more quantitatively by the decrease in the peaks of an energy loss spectrum. In the latter case the exposure, EC, to decrease the peak intensity from IO to I’O can be related to the molecular dissociation cross-section, σD, by EC = ℓn(IO /I’O) /ℓD. Qu ntitative data on damage cross-sections are just being reported, However, the microscopist needs to know the explicit dependence of damage on: (1) the molecular properties, (2) the density and characteristics of the molecular film and that of the support film, if any, (3) the temperature of the molecular film and (4) certain characteristics of the electron microscope used


Author(s):  
H. Tochigi ◽  
H. Uchida ◽  
S. Shirai ◽  
K. Akashi ◽  
D. J. Evins ◽  
...  

A New High Excitation Objective Lens (Second-Zone Objective Lens) was discussed at Twenty-Sixth Annual EMSA Meeting. A new commercially available Transmission Electron Microscope incorporating this new lens has been completed.Major advantages of the new instrument allow an extremely small beam to be produced on the specimen plane which minimizes specimen beam damages, reduces contamination and drift.


Author(s):  
S. Saito ◽  
H. Todokoro ◽  
S. Nomura ◽  
T. Komoda

Field emission scanning electron microscope (FESEM) features extremely high resolution images, and offers many valuable information. But, for a specimen which gives low contrast images, lateral stripes appear in images. These stripes are resulted from signal fluctuations caused by probe current noises. In order to obtain good images without stripes, the fluctuations should be less than 1%, especially for low contrast images. For this purpose, the authors realized a noise compensator, and applied this to the FESEM.Fig. 1 shows an outline of FESEM equipped with a noise compensator. Two apertures are provided gust under the field emission gun.


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