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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

how does Neisseria cause meningitis
Nyibol Reply
what is microbiologist
Muhammad Reply
what is errata
Muhammad
is the branch of biology that deals with the study of microorganisms.
Ntefuni Reply
What is microbiology
Mercy Reply
studies of microbes
Louisiaste
when we takee the specimen which lumbar,spin,
Ziyad Reply
How bacteria create energy to survive?
Muhamad Reply
Bacteria doesn't produce energy they are dependent upon their substrate in case of lack of nutrients they are able to make spores which helps them to sustain in harsh environments
_Adnan
But not all bacteria make spores, l mean Eukaryotic cells have Mitochondria which acts as powerhouse for them, since bacteria don't have it, what is the substitution for it?
Muhamad
they make spores
Louisiaste
what is sporadic nd endemic, epidemic
Aminu Reply
the significance of food webs for disease transmission
Abreham
food webs brings about an infection as an individual depends on number of diseased foods or carriers dully.
Mark
explain assimilatory nitrate reduction
Esinniobiwa Reply
Assimilatory nitrate reduction is a process that occurs in some microorganisms, such as bacteria and archaea, in which nitrate (NO3-) is reduced to nitrite (NO2-), and then further reduced to ammonia (NH3).
Elkana
This process is called assimilatory nitrate reduction because the nitrogen that is produced is incorporated in the cells of microorganisms where it can be used in the synthesis of amino acids and other nitrogen products
Elkana
Examples of thermophilic organisms
Shu Reply
Give Examples of thermophilic organisms
Shu
advantages of normal Flora to the host
Micheal Reply
Prevent foreign microbes to the host
Abubakar
they provide healthier benefits to their hosts
ayesha
They are friends to host only when Host immune system is strong and become enemies when the host immune system is weakened . very bad relationship!
Mark
what is cell
faisal Reply
cell is the smallest unit of life
Fauziya
cell is the smallest unit of life
Akanni
ok
Innocent
cell is the structural and functional unit of life
Hasan
is the fundamental units of Life
Musa
what are emergency diseases
Micheal Reply
There are nothing like emergency disease but there are some common medical emergency which can occur simultaneously like Bleeding,heart attack,Breathing difficulties,severe pain heart stock.Hope you will get my point .Have a nice day ❣️
_Adnan
define infection ,prevention and control
Innocent
I think infection prevention and control is the avoidance of all things we do that gives out break of infections and promotion of health practices that promote life
Lubega
Heyy Lubega hussein where are u from?
_Adnan
en français
Adama
which site have a normal flora
ESTHER Reply
Many sites of the body have it Skin Nasal cavity Oral cavity Gastro intestinal tract
Safaa
skin
Asiina
skin,Oral,Nasal,GIt
Sadik
How can Commensal can Bacteria change into pathogen?
Sadik
How can Commensal Bacteria change into pathogen?
Sadik
all
Tesfaye
by fussion
Asiina
what are the advantages of normal Flora to the host
Micheal
what are the ways of control and prevention of nosocomial infection in the hospital
Micheal
what is inflammation
Shelly Reply
part of a tissue or an organ being wounded or bruised.
Wilfred
what term is used to name and classify microorganisms?
Micheal Reply
Binomial nomenclature
adeolu
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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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