Comments of JSC SRC Progress to the Article “Limiting Instrumental Resolution of the Resurs-P Spacecraft (No. 1, 2, 3)” by K.N. Sviridov (journal “Rocket-Space Device Engineering and Information Systems”, 2017, Vol. 4, Iss. 2, pp. 20–28)

High magnification imaging of macromolecules on metal coated biological specimens is limited only by wet preparation procedures since recently obtained instrumental resolution allows visualization of topographic structures as smal l as 1-2 nm. Details of such dimensions may be visualized if continuous metal films with a thickness of 2 nm or less are applied. Such thin films give sufficient contrast in TEM as well as in SEM (SE-I image mode). The requisite increase in electrical conductivity for SEM of biological specimens is achieved through the use of ligand mediated wet osmiuum impregnation of the specimen before critical point (CP) drying. A commonly used ligand is thiocarbohvdrazide (TCH), first introduced to TEM for en block staining of lipids and glvcomacromolecules with osmium black. Now TCH is also used for SEM. However, after ligand mediated osinification nonspecific osmium black precipitates were often found obscuring surface details with large diffuse aggregates or with dense particular deposits, 2-20 nm in size. Thus, only low magnification work was considered possible after TCH appl ication.

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High-Resolution Cryo-Electron Microscopy of Biological Macromolecules

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100179385 ◽

1990 ◽

Vol 48 (1) ◽

pp. 126-127

Author(s):

Yoshinori Fujiyoshi

Keyword(s):

Electron Microscopy ◽

High Resolution ◽

Diffraction Pattern ◽

Superfluid Helium ◽

Copper Phthalocyanine ◽

Optical Diffraction ◽

Irradiation Damage ◽

Biological Macromolecules ◽

Cross Sectional ◽

Instrumental Resolution

The resolution of direct images of biological macromolecules is normally restricted to far less than 0.3 nm. This is not due instrumental resolution, but irradiation damage. The damage to biological macromolecules may expect to be reduced when they are cooled to a very low temperature. We started to develop a new cryo-stage for a high resolution electron microscopy in 1983, and successfully constructed a superfluid helium stage for a 400 kV microscope by 1986, whereby chlorinated copper-phthalocyanine could be photographed to a resolution of 0.26 nm at a stage temperature of 1.5 K. We are continuing to develop the cryo-microscope and have developed a cryo-microscope equipped with a superfluid helium stage and new cryo-transfer device.The New cryo-microscope achieves not only improved resolution but also increased operational ease. The construction of the new super-fluid helium stage is shown in Fig. 1, where the cross sectional structure is shown parallel to an electron beam path. The capacities of LN2 tank, LHe tank and the pot are 1400 ml, 1200 ml and 3 ml, respectively. Their surfaces are placed with gold to minimize thermal radiation. Consumption rates of liquid nitrogen and liquid helium are 170 ml/hour and 140 ml/hour, respectively. The working time of this stage is more than 7 hours starting from full LN2 and LHe tanks. Instrumental resolution of our cryo-stage cooled to 4.2 K was confirmed to be 0.20 nm by an optical diffraction pattern from the image of a chlorinated copper-phthalocyanine crystal. The image and the optical diffraction pattern are shown in Fig. 2 a, b, respectively.

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Observation of GaAs/AlAs Superlattice Structures in both Secondary and Backscattcred Electron Imaging Modes with an Ultrahigh Resolution Scanning Electron Microscope

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010018077x ◽

1990 ◽

Vol 48 (1) ◽

pp. 404-405

Author(s):

K. Ogura ◽

A. Ono ◽

S. Franchi ◽

P.G. Merli ◽

A. Migliori

Keyword(s):

Gaas Layer ◽

Objective Lens ◽

Superlattice Structure ◽

Backscattered Electron ◽

Instrumental Resolution ◽

Electron Microscopes ◽

Superlattice Structures ◽

Electron Imaging ◽

Scanning Electron Microscopes ◽

Scanning Electron

In the last few years the development of Scanning Electron Microscopes (SEM), equipped with a Field Emission Gun (FEG) and using in-lens specimen position, has allowed a significant improvement of the instrumental resolution . This is a result of the fine and bright probe provided by the FEG and by the reduced aberration coefficients of the strongly excited objective lens. The smaller specimen size required by in-lens instruments (about 1 cm, in comparison to 15 or 20 cm of a conventional SEM) doesn’t represent a serious limitation in the evaluation of semiconductor process techniques, where the demand of high resolution is continuosly increasing. In this field one of the more interesting applications, already described (1), is the observation of superlattice structures.In this note we report a comparison between secondary electron (SE) and backscattered electron (BSE) images of a GaAs / AlAs superlattice structure, whose cross section is reported in fig. 1. The structure consist of a 3 nm GaAs layer and 10 pairs of 7 nm GaAs / 15 nm AlAs layers grown on GaAs substrate. Fig. 2, 3 and 4 are SE images of this structure made with a JEOL JSM 890 SEM operating at an accelerating voltage of 3, 15 and 25 kV respectively. Fig. 5 is a 25 kV BSE image of the same specimen. It can be noticed that the 3nm layer is always visible and that the 3 kV SE image, in spite of the poorer resolution, shows the same contrast of the BSE image. In the SE mode, an increase of the accelerating voltage produces a contrast inversion. On the contrary, when observed with BSE, the layers of GaAs are always brighter than the AlAs ones , independently of the beam energy.

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