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Introduction

The scanning electron microscope (SEM) is a very useful imaging technique that utilized a beam of electrons to acquire high magnification images of specimens. Very similar to the transmission electron microscope (TEM), the SEM maps the reflected electrons and allows imaging of thick (~mm) samples, whereas the TEM requires extremely thin specimens for imaging; however, the SEM has lower magnifications. Although both SEM and TEM use an electron beam, the image is formed very differently and users should be aware of when each microscope is advantageous.

Microscopy physics

Image formation

All microscopes serve to enlarge the size of an object and allow people to view smaller regions within the sample. Microscopes form optical images and although instruments like the SEM have extremely high magnifications, the physics of the image formation are very basic. The simplest magnification lens can be seen in [link] . The formula for magnification is shown in [link] , where M is magnification, f is focal length, u is the distance between object and lens, and v is distance from lens to the image.

M = f u f = v f f size 12{M= { {f} over {u - f} } = { {v - f} over {f} } } {}
Basic microscope diagram illustrating inverted image and distances u , f , and v .

Multistage microscopes can amplify the magnification of the original object even more as shown in [link] . Where magnification is now calculated from [link] , where f 1 , f 2 are focal distances with respect to the first and second lens and v 1 , v 2 are the distances from the lens to the magnified image of first and second lens, respectively.

M = ( v 1 f 1 ) ( v 2 f 2 ) f 1 f 2 size 12{M= { { \( v rSub { size 8{1} } - f rSub { size 8{1} } \) \( v rSub { size 8{2} } - f rSub { size 8{2} } \) } over {f rSub { size 8{1} } f rSub { size 8{2} } } } } {}
A schematic diagram of the optics used in a multistage microscope.

In reality, the objects we wish to magnify need to be illuminated. Whether or not the sample is thin enough to transmit light divides the microscope into two arenas. SEM is used for samples that do not transmit light, whereas the TEM (transmission electron microscope) requires transparent samples. Due to the many frequencies of light from the introduced source, a condenser system is added to control the brightness and narrow the range of viewing to reduce aberrations, which distort the magnified image.

Electron microscopes

Microscope images can be formed instantaneous (as in the optical microscope or TEM) or by rastering (scanning) a beam across the sample and forming the image point-by-point. The latter is how SEM images are formed. It is important to understand the basic principles behind SEM that define properties and limitations of the image.

Resolution

The resolution of a microscope is defined as the smallest distance between two features that can be uniquely identified (also called resolving power). There are many limits to the maximum resolution of the SEM and other microscopes, such as imperfect lenses and diffraction effects. Each single beam of light, once passed through a lens, forms a series of cones called an airy ring (see [link] ). For a given wavelength of light, the central spot size is inversely proportional to the aperture size (i.e., large aperture yields small spot size) and high resolution demands a small spot size.

Questions & Answers

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Anatomy is the study of the structure of the body, while physiology is the study of the function of the body. Anatomy looks at the body's organs and systems, while physiology looks at how those organs and systems work together to keep the body functioning.
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Source:  OpenStax, Physical methods in chemistry and nano science. OpenStax CNX. May 05, 2015 Download for free at http://legacy.cnx.org/content/col10699/1.21
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