Magnification in Modern Scanning Electron Microscopes
Magnification in Modern Scanning Electron Microscopes
Application Note
Introduction
From its earliest inception, the Scanning Electron Microscope (SEM) has been widely used
as an imaging tool. It produces images by raster scanning an electron beam over a region of
interest on a sample. The SEM allows for the visualization of features too small to resolve by
the unaided human eye. Early SEM images were analog images which were preserved on 4 x
5 Polaroid or Kodak film. The proliferation of high speed digital electronics has revolutionized
SEM, whereby everything from digital scan control to digital acquisition, to archival of digital
images is not only common but expected by default on modern SEMs. Since this digital rev-
olution, digital images are displayed on everything from desktop computer monitors to large
projection screens and printed at various pixel densities on a wide variety of paper sizes.
Once the image is projected or reproduced, the size of the image, and therefore the magnifi-
cation depends on the scale at which the image is viewed. Hence, the original magnification
value when the image was collected is irrelevant at best and very misleading at worst. By
comparison, another raster scanning microscope which produces digital images, the Atomic
Force Microscope (AFM), addresses the issues of magnification by referencing the scan size
opposed to magnification. For any digital image, the scale of the image, i.e. nanometers per
pixel for microscopes or kilometers per pixel for satellite images, is a fundamental property
not the magnification. In addition to the image scale other contributing factors to what can be
visualized and measured are empty magnification and pixel resolution.
With AFM which has been a digital microscope since its introduction, magnification is a
non-issue because the images are always referenced to the scan size (the actual area on the
sample that was scanned). Scan size is a very useful, display-independent way to view and
analyze digital images. Given the many advantages digital images and their widespread use in
microscopy, there is a compelling need to standardize on scan size when discussing the scale
of features observed in digital images.
03 | Keysight | Why Magnification is Irrelevant in Modern Scanning Electron Microscopes - Application Note
Figure 1. Analog scan generation to create a 1024 x 1024 image. Figure 2. Digital scan generation to create a 1024 x 1024 image.
The magnification was well controlled because the ratio of film size to internal CRT screen
size was fixed. Therefore the original magnification value was directly linked to the photo-
graph. With the advent of digital imaging and the variety of display formats, this link has
been broken.
Figure 3. Particle size distribution calculation on threshold Figure 4. Image analysis showing darker phase comprises
image. 60% of the sample.
An advantage with digital image data is the intensity data can easily be normalized for
example, with the typical 8 bit gray scale SEM image, the darkest pixel is set to 0 and
the brightest is set to 255. Normalization is sometimes referred to as ABC (auto bright-
ness and contrast) and it allows for convenient storage and display of the data on the
computer monitor without the need to worry about film speed as in analog SEM images.
However, with normalized digital images it is not appropriate to compare absolute image
brightness between images, as was done with controlled exposure photographs.
With the scan size, and therefore the scale of the image known, digital image analysis or
processing affords increasing sophistication in what can be analyzed in the collected im-
age data. There are many sophisticated software programs for manipulating and analyz-
ing digital images. With digital images, analyses like particle size distribution (Figure 3),
average fiber length, or area ratio of one phase to another in a multicomponent system
can be done easily (Figure 4).
05 | Keysight | Why Magnification is Irrelevant in Modern Scanning Electron Microscopes - Application Note
Figure 6. Computer simulation of a 100nm electron beam scanning various geometric shapes
and their resulting image profiles.
Figure 5. Computer simulation of a 100 nm electron Figure 7. Computer simulation of a 10nm electron beam scanning various geometric shapes
beam. and their resulting image profiles.
Although electron beam diameter is the determining factor of resolution in the ideal
case, in practice there are the following subordinate factors, sample preparation and
surface roughness, atomic mass and chemical composition of the sample, beam inten-
sity, accelerating voltage, scanning speed, working distance, aberrations and hysteresis
in the electron optics, and interaction volume of the electron beam with the sample.
Because the electron beam diameter is the dominant factor, for the examples below the
contribution of the subordinate factors were assumed to be negligible.
In Figure 6 we see how a 100nm electron beam interacts with a triangle, parallelogram,
pentagon, and a hexagon and the representation of how the shapes would nominally be
represented on the computer monitor. With a much smaller, 10nm electron beam inter-
acting with the same geometric shapes we see in Figure 7 that the resulting images give
a much more accurate representation of the original shape.
06 | Keysight | Why Magnification is Irrelevant in Modern Scanning Electron Microscopes - Application Note
Pixel Resolution
For the analog SEM the images were recorded on film, so the exposure and grain size of the film
determined the smallest features which could be imaged. For the digital SEM pixel resolution
determines the smallest features which could be imaged. Each pixel in a digital image contains
just one element of information, i.e. a gray level from 0 (black) to 255 (white). The smallest feature
which can be resolved is therefore linked to the pixel size, as seen in Figures 8–13.
Figure 8. SEM image defined by 28 x 28 pixels. Figure 9. SEM image defined by 56 x 56 pixels.
Figure 10. SEM image defined by 84 x 84 pixels. Figure 11. SEM image defined by 112 x 112 pixels.
Figure 12. SEM image defined by 140 x 140 pixels. Figure 13. SEM image defined by 2048 x 2048 pixels.
07 | Keysight | Why Magnification is Irrelevant in Modern Scanning Electron Microscopes - Application Note
Figure 14. Sketch of Empty Magnification. If an object is “magnified” beyond the resolution based on the electron
beam shape it results in the equivalent to digital zoom, i.e. 1 pixel of significant data is divided into 4 identical pixels.
In Figure 8 it is difficult to discern any sample features. As the pixel resolution, pixel density
per area scanned on the sample, is increased to even a modest number, 84 x 84 pixels in
Figure 10, the gold islands on carbon test sample can now be recognized, but not suffi-
ciently for making accurate measurements. Ultimately the best image, especially for mak-
ing dimensional measurements, is seen in Figure 13 where the pixel density is 2048 x 2048.
Empty Magnification
The artifact of empty magnification is present in both analog and digital SEMs. The rea-
soning behind the term empty magnification is that the magnification can be increased
such that it exceeds the resolution based on the electron beam size; therefore the image
is empty of significant information. With the digital SEM, although the increase in pixel
resolution improves the image, it should be noted that the ultimate resolution is defined
by the shape of the electron beam, not the number of pixels. If we add a meaningful
size scale to the geometric shapes in Figure 6, Figure 7, we see that no matter what
instrument magnification or pixel density we use, the images generated with the 100nm
electron beam will not accurately reproduce the original shapes.
Another way of demonstrating empty magnification is shown in Figure 14, where the
simulated object on the left is the smallest feature that can be resolved and then either the
instrument magnification is increased or the pixel density is increased. The result on the
right side of Figure 14, no new information is generated 1 unit of significant data is divided
into 4 identical units. The result is the information content is the same, there are just more
pixels. The same principal is true for increasing the instrument magnification such that the
electron beam becomes large compared to the pixel or feature size.
Therefore just because a SEM can be set to 1,000,000x magnification does not mean the
resulting image scale of 0.2nm per pixel translates into usable magnification. As seen in
Figure 6, with a 100nm electron beam size it does not matter how many pixels are used
or how small a scan size is used the resulting image does not accurately represent the
original sample. However, if the electron beam is small, as seen in Figure 7, increasing
the pixel resolution can be helpful in visualizing small features, i.e. the corners of the
triangle shape. This is similar to digital zoom on a digital camera.
Figure 15. AC mode AFM image of Celgard polymer 3 µm x 3 µm scan size. Figure 16. SEM image of Celgard polymer 2.5 µm x 2.5 µm scan size.
Imaged on Keysight 5500 AFM. Imaged on Keysight 8500 FE-SEM.
Conclusions
In modern SEMs most of the signals are handled digitally and for digital images, the
scale of the image is the fundamental property not the magnification. Therefore, the
original “magnification” value when the image was collected is irrelevant at best and
very misleading at worst. With commercial SEM instruments, if scan size as opposed to
magnification was used it would readily allow side by side comparison of images from
different instruments as well as easier comparison to AFM images. With the scan size,
and therefore the scale of the image known, digital image processing delivers increasing
sophistication in what can be analyzed in the collected image data.
09 | Keysight | Why Magnification is Irrelevant in Modern Scanning Electron Microscopes - Application Note
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