Low-voltage high-resolution SEM of biological samples

1989 ◽  
Vol 47 ◽  
pp. 72-73
Author(s):  
James B. Pawley
Keyword(s):  
Secondary Electron ◽  
Low Voltage ◽  
Focal Length ◽  
Semiconductor Industry ◽  
Focal Length Lens ◽  
Small Probe ◽  
Beam Voltage ◽  
Recent Developments ◽  
Topographical Feature ◽  
Beam Damage

There are three reasons for the recent upsurge of interest in using the SEM at a beam voltage (Vo) around 1 kV (LVSEM). The most common one arises from applications in the semiconductor industry and emphasizes the reduction in charging artifacts and in subsurface beam damage obtainable at low VoThe second reason derives from the belief that, given instrumentation capable of producing a sufficiently small probe, the increased contrast in the topographic component of the secondary electron signal will permit an important improvement in real topographic spatial resolution.The third reason is that recent developments in instrumentation have shown that by coupling a cold field-emission source with a short focal length lens, it is indeed possible to obtain small probe diameters at low voltage. Although they are considerably larger than probes obtained at higher voltage, they are nonetheless smaller than the smallest topographical feature yet imaged in the secondary electron (SE) mode. (Fig 1)

scholarly journals Low-voltage scanning electron microscopy

1983 ◽  
Vol 41 ◽  
pp. 488-491
Author(s):  
J. B. Pawley ◽  
M. P. Winters
Keyword(s):  
Electron Probe ◽  
Current Flow ◽  
Low Voltage ◽  
Semiconductor Industry ◽  
X Rays ◽  
Probe Diameter ◽  
Small Probe ◽  
Important Improvement ◽  
Voltage Scanning ◽  
Beam Damage

There are two reasons for the renewed interest in using the SEM at beam voltages, Vb, around 1kV (LVSEM). The most common one arises from applications in the semiconductor industry and emphasizes the reduction in charging artifacts and in subsurface beam damage. The second reason postulates that the increased contrast in the topographic component of the secondary electron signal will permit an important improvement in topographic spatial resolution if only a sufficiently small probe diameter can be obtained. We shall treat these two areas separately and then mention some of the strategies that have been adopted to make LVSEM work.Surfaces are very important in the manufacture of modern semiconductor devices and the ability of the electron probe to induce current flow (EBIC), to detect variations in surface voltage, Vs, and to excite characteristic x rays in addition to its ability to image topography in an easily understandable way guaranteed the SEM a major role in programs of semiconductor development and failure analysis. There were two problems, however, charging and beam-induced damage to the specimen.

High-Resolution stereo imaging of frozen-hydrated biological samples by Cryo-SEM

1992 ◽  
Vol 50 (2) ◽  
pp. 1280-1281
Author(s):  
Ya Chen ◽  
Paul Walther
Keyword(s):  
High Resolution ◽  
Focal Length ◽  
Crystal Formation ◽  
Ice Crystal ◽  
Stereo Imaging ◽  
Focal Length Lens ◽  
Fine Grain ◽  
Beam Voltage ◽  
Resolution Imaging ◽  
Beam Damage

The introduction of cryo-techniques to SEM allowed the biologist to examine frozen-hydrated specimens at low temperatures on a cryo-stage in the SEM. The problems associated with conventional cryo-SEM included ice crystal formation caused by low cooling rate, poor resolution, limited magnification (<10 kX), specimen contamination, beam damage and charging. Recently, the effectiveness of this technique has been improved because a number of new cryo-preparation instruments have become commercially available. The high pressure freezer permits cryo-fixation of large biological specimens (diameter up to about 1mm) without ice crystal artifacts. Cryo-coating units permit fine-grain metal coating of fractured specimens. In addition, microscopes combining a short focal length lens with a fieldemission (FE) source permit high resolution imaging at low beam voltage (Vo). However, for high resolution cryo-SEM, especially for producing stereo pictures, the artifacts caused by electron irradiation remain a critical problem.The purpose of this work was to evaluate the extent to which beam damage varies with Vo, and to determine the most appropriate Vo for stereo imaging.

SEM at Low Beam Voltage

1984 ◽  
Vol 42 ◽  
pp. 440-443
Author(s):  
James Pawley
Keyword(s):  
Low Voltage ◽  
Probe Diameter ◽  
Small Probe ◽  
Beam Voltage ◽  
Practical Constraints

Operation of the SEM with V0 = l-3kV (LVSEM) was early recognized to reduce charging artefacts and increase topographic contrast. This early promise was not pursued because several theoretical and practical considerations made it difficult to produce a small probe diameter (d0) at low voltage. Recently, the necessity of using low V0 to image uncoated semiconductors without damaging them has prompted a re-evaluation of LVSEM. This re-evaluation has taken the form of efforts to eliminate the practical constraints and to alleviate the theoretical ones. In the process, some heretofore neglected theoretical advantages of LVSEM have emerged. These problems and possibilities will now be discussed in more detail.

Recent developments in low-voltage SEM of polymers

1993 ◽  
Vol 51 ◽  
pp. 866-867
Author(s):  
S.J. Krause ◽  
W.W. Adams
Keyword(s):  
Low Voltage ◽  
Chromatic Aberration ◽  
Thermal Electron ◽  
Analytical Technique ◽  
Recent Developments ◽  
Beam Currents ◽  
New Generation ◽  
First Time ◽  
Voltage Scanning ◽  
Beam Damage

Over the past decade low voltage scanning electron microscopy (LVSEM) of polymers has evolved from an interesting curiosity to a powerful analytical technique. This development has been driven by improved instrumentation and in particular, reliable field emission gun (FEG) SEMs. The usefulness of LVSEM has also grown because of an improved theoretical and experimental understanding of sample-beam interactions and by advances in sample preparation and operating techniques. This paper will review progress in polymer LVSEM and present recent results and developments in the field.In the early 1980s a new generation of SEMs produced beam currents that were sufficient to allow imaging at low voltages from 5keV to 0.5 keV. Thus, for the first time, it became possible to routinely image uncoated polymers at voltages below their negative charging threshold, the "second crossover", E2 (Fig. 1). LVSEM also improved contrast and reduced beam damage in sputter metal coated polymers. Unfortunately, resolution was limited to a few tenths of a micron due to the low brightness and chromatic aberration of thermal electron emission sources.

Improvements in the electron probe forming capabilities and x-ray hole counts in the Philips TEM

1983 ◽  
Vol 41 ◽  
pp. 376-377
Author(s):  
Richard L. McConville
Keyword(s):  
Field Strength ◽  
Electron Probe ◽  
Spherical Aberration ◽  
Focal Length ◽  
Objective Lens ◽  
Parallel Beam ◽  
Tilt Axis ◽  
X Ray ◽  
Small Probe ◽  
Specimen Height

A second generation twin lens has been developed. This symmetrical lens with a wider bore, yet superior values of chromatic and spherical aberration for a given focal length, retains both eucentric ± 60° tilt movement and 20°x ray detector take-off angle at 90° to the tilt axis. Adjust able tilt axis height, as well as specimen height, now ensures almost invariant objective lens strengths for both TEM (parallel beam conditions) and STEM or nano probe (focused small probe) modes.These modes are selected through use of an auxiliary lens situ ated above the objective. When this lens is on the specimen is illuminated with a parallel beam of electrons, and when it is off the specimen is illuminated with a focused probe of dimensions governed by the excitation of the condenser 1 lens. Thus TEM/STEM operation is controlled by a lens which is independent of the objective lens field strength.

SEM analysis of DC magnetron sputtered beryllium films

1988 ◽  
Vol 46 ◽  
pp. 960-961
Author(s):  
E. F. Lindsey ◽  
C. W. Price ◽  
E. L. Pierce ◽  
E. J. Hsieh
Keyword(s):  
Fine Structure ◽  
Electron Emission ◽  
Secondary Electron ◽  
Low Voltage ◽  
Ion Beam ◽  
Secondary Electron Emission ◽  
Sem Analysis ◽  
Fused Silica ◽  
Grain Structure ◽  
Silica Substrate

Columnar structures produced by DC magnetron sputtering can be altered by using RF biased sputtering or by exposing the film to nitrogen pulses during sputtering, and these techniques are being evaluated to refine the grain structure in sputtered beryllium films deposited on fused silica substrates. Beryllium is brittle, and fractures in sputtered beryllium films tend to be intergranular; therefore, a convenient technique to analyze grain structure in these films is to fracture the coated specimens and examine them in an SEM. However, fine structure in sputtered deposits is difficult to image in an SEM, and both the low density and the low secondary electron emission coefficient of beryllium seriously compound this problem. Secondary electron emission can be improved by coating beryllium with Au or Au-Pd, and coating also was required to overcome severe charging of the fused silica substrate even at low voltage. The coating structure can obliterate much of the fine structure in beryllium films, but reasonable results were obtained by using the high-resolution capability of an Hitachi S-800 SEM and either ion-beam coating with Au-Pd or carbon coating by thermal evaporation.

High-Resolution, Low-Voltage SEM of Cell Wall Regeneration of Yeast Shizosaccharomyces Pombe Protoplasts

1988 ◽  
Vol 46 ◽  
pp. 208-211
Author(s):  
M. Osumi ◽  
N. Yamada ◽  
T. Nagatani
Keyword(s):  
Electron Microscope ◽  
Scanning Electron Microscope ◽  
Low Voltage ◽  
Cell Wall Regeneration ◽  
The Past ◽  
The Poor ◽  
Probe Size ◽  
Beam Damage ◽  
Biological Field ◽  
Scanning Electron

Even though many early workers had suggested the use of lower voltages to increase topographic contrast and to reduce specimen charging and beam damage, we did not usually operate in the conventional scanning electron microscope at low voltage because of the poor resolution, especially of bioligical specimens. However, the development of the “in-lens” field emission scanning electron microscope (FESEM) has led to marked inprovement in resolution, especially in the range of 1-5 kV, within the past year. The probe size has been cumulated to be 0.7nm in diameter at 30kV and about 3nm at 1kV. We have been trying to develop techniques to use this in-lens FESEM at low voltage (LVSEM) for direct observation of totally uncoated biological specimens and have developed the LVSEM method for the biological field.

Low-voltage, high-resolution scanning electron microscopy of polymers

1987 ◽  
Vol 45 ◽  
pp. 466-467 ◽  
Author(s):  
S. J. Krause ◽  
W.W. Adams ◽  
S. Kumar ◽  
T. Reilly ◽  
T. Suziki
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Low Voltage ◽  
Chromatic Aberration ◽  
Image Resolution ◽  
Resolution Limit ◽  
Crossover Point ◽  
New Generation ◽  
Beam Damage ◽  
Scanning Electron

Scanning electron microscopy (SEM) of polymers at routine operating voltages of 15 to 25 keV can lead to beam damage and sample image distortion due to charging. Imaging polymer samples with low accelerating voltages (0.1 to 2.0 keV), at or near the “crossover point”, can reduce beam damage, eliminate charging, and improve contrast of surface detail. However, at low voltage, beam brightness is reduced and image resolution is degraded due to chromatic aberration. A new generation of instruments has improved brightness at low voltages, but a typical SEM with a tungsten hairpin filament will have a resolution limit of about 100nm at 1keV. Recently, a new field emission gun (FEG) SEM, the Hitachi S900, was introduced with a reported resolution of 0.8nm at 30keV and 5nm at 1keV. In this research we are reporting the results of imaging coated and uncoated polymer samples at accelerating voltages between 1keV and 30keV in a tungsten hairpin SEM and in the Hitachi S900 FEG SEM.

Linewidth measurement using the low voltage SEM

1986 ◽  
Vol 44 ◽  
pp. 652-653
Author(s):  
M.G. Rosenfield
Keyword(s):  
Electron Microscope ◽  
Scanning Electron Microscope ◽  
Integrated Circuits ◽  
Secondary Electron ◽  
Low Voltage ◽  
Fabrication Process ◽  
Critical Dimensions ◽  
Linewidth Measurement ◽  
Threshold Technique ◽  
Scanning Electron

Minimum feature sizes in experimental integrated circuits are approaching 0.5 μm and below. During the fabrication process it is usually necessary to be able to non-destructively measure the critical dimensions in resist and after the various process steps. This can be accomplished using the low voltage SEM. Submicron linewidth measurement is typically done by manually measuring the SEM micrographs. Since it is desirable to make as many measurements as possible in the shortest period of time, it is important that this technique be automated.Linewidth measurement using the scanning electron microscope is not well understood. The basic intent is to measure the size of a structure from the secondary electron signal generated by that structure. Thus, it is important to understand how the actual dimension of the line being measured relates to the secondary electron signal. Since different features generate different signals, the same method of relating linewidth to signal cannot be used. For example, the peak to peak method may be used to accurately measure the linewidth of an isolated resist line; but, a threshold technique may be required for an isolated space in resist.

scholarly journals Quantitative material analysis using secondary electron energy spectromicroscopy

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
W. Han ◽  
M. Zheng ◽  
A. Banerjee ◽  
Y. Z. Luo ◽  
L. Shen ◽  
...  
Keyword(s):  
Electron Energy ◽  
Close Agreement ◽  
Secondary Electron ◽  
Low Voltage ◽  
High Accuracy ◽  
Material Analysis ◽  
Auger Microscopy ◽  
New Applications ◽  
Voltage Scanning ◽  
Scanning Electron

AbstractThis paper demonstrates how secondary electron energy spectroscopy (SEES) performed inside a scanning electron microscope (SEM) can be used to map sample atomic number and acquire bulk valence band density of states (DOS) information at low primary beam voltages. The technique uses an electron energy analyser attachment to detect small changes in the shape of the scattered secondary electron (SE) spectrum and extract out fine structure features from it. Close agreement between experimental and theoretical bulk valance band DOS distributions was obtained for six different test samples, where the normalised root mean square deviation ranged from 2.7 to 6.7%. High accuracy levels of this kind do not appear to have been reported before. The results presented in this paper point towards SEES becoming a quantitative material analysis companion tool for low voltage scanning electron microscopy (LVSEM) and providing new applications for Scanning Auger Microscopy (SAM) instruments.

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