A high-resolution membrane probe

Author(s):  
James F. Hainfeld ◽  
James J. Lipka ◽  
F. Elsie Quaite

Labeling the lipids of membranes for E.M. is frequently done by using radioactive phospholipids. Membrane dynamics can then be followed by autoradiography. Unfortunately, it takes approximately 6 months to expose the emulsion due to the low specific activity of the labels. Also, the resolution is limited to about 0.2 μm by this technique.An alternative is to develop a phospholipid-like molecule that is electron dense and could be visualized directly. Several such molecules have been synthesized and one is shown in Fig. 1. They have a phosphotungstate “head” group that has a 4 (-) charge and is hydrophilic and an aliphatic “tail” of varying length that is hydrophobic. The tungstate cluster contains 11 tungsten atoms in the “Keggin” structure with a 1.0 nm diameter and has previously been studied in the electron microscope. It was shown to be almost completely resistant to beam induced mass loss.

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.


Author(s):  
H. Rose

The scanning transmission electron microscope offers the possibility of utilizing inelastically scattered electrons. Use of these electrons in addition to the elastically scattered electrons should reduce the scanning time (dose) Which is necessary to keep the quantum noise below a certain level. Hence it should lower the radiation damage. For high resolution, Where the collection efficiency of elastically scattered electrons is small, the use of Inelastically scattered electrons should become more and more favorable because they can all be detected by means of a spectrometer. Unfortunately, the Inelastic scattering Is a non-localized interaction due to the electron-electron correlation, occurring predominantly at the circumference of the atomic electron cloud.


Author(s):  
J. P. Langmore ◽  
N. R. Cozzarelli ◽  
A. V. Crewe

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.


Author(s):  
David Joy ◽  
James Pawley

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).


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):  
M.K. Lamvik ◽  
D.A. Kopf ◽  
S.D. Davilla ◽  
J.D. Robertson

Last year we reported1 that there is a striking reduction in the rate of mass loss when a specimen is observed at liquid helium temperature. It is important to determine whether liquid helium temperature is significantly better than liquid nitrogen temperature. This requires a good understanding of mass loss effects in cold stages around 100K.


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