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Certain materials can refract nonvisible forms of EMR and, in effect, transform them into visible light. Certain fluorescent dyes, for instance, absorb ultraviolet or blue light and then use the energy to emit photons of a different color, giving off light rather than simply vibrating. This occurs because the energy absorption causes electrons to jump to higher energy states, after which they then almost immediately fall back down to their ground states, emitting specific amounts of energy as photons. Not all of the energy is emitted in a given photon, so the emitted photons will be of lower energy and, thus, of lower frequency than the absorbed ones. Thus, a dye such as Texas red may be excited by blue light, but emit red light; or a dye such as fluorescein isothiocyanate (FITC) may absorb (invisible) high-energy ultraviolet light and emit green light ( [link] ). In some materials, the photons may be emitted following a delay after absorption; in this case, the process is called phosphorescence . Glow-in-the-dark plastic works by using phosphorescent material.

An image shows a large cell in the foreground and other cells further in the background. Each cell has an irregular shape with a large blue circle in the center. Green lines surround the blue circle and span out towards the edges of the cell. The rest of the cell is red with a bright red edge. The background of the image is black.
The fluorescent dyes absorbed by these bovine pulmonary artery endothelial cells emit brilliant colors when excited by ultraviolet light under a fluorescence microscope. Various cell structures absorb different dyes. The nuclei are stained blue with 4’,6-diamidino-2-phenylindole (DAPI); microtubles are marked green by an antibody bound to FITC; and actin filaments are labeled red with phalloidin bound to tetramethylrhodamine (TRITC). (credit: National Institutes of Health)
  • Which has a higher frequency: red light or green light?
  • Explain why dispersion occurs when white light passes through a prism.
  • Why do fluorescent dyes emit a different color of light than they absorb?

Magnification, resolution, and contrast

Microscopes magnify images and use the properties of light to create useful images of small objects. Magnification is defined as the ability of a lens to enlarge the image of an object when compared to the real object. For example, a magnification of 10⨯ means that the image appears 10 times the size of the object as viewed with the naked eye.

Greater magnification typically improves our ability to see details of small objects, but magnification alone is not sufficient to make the most useful images. It is often useful to enhance the resolution of objects: the ability to tell that two separate points or objects are separate. A low-resolution image appears fuzzy, whereas a high-resolution image appears sharp. Two factors affect resolution. The first is wavelength. Shorter wavelengths are able to resolve smaller objects; thus, an electron microscope has a much higher resolution than a light microscope, since it uses an electron beam with a very short wavelength, as opposed to the long-wavelength visible light used by a light microscope. The second factor that affects resolution is numerical aperture , which is a measure of a lens’s ability to gather light. The higher the numerical aperture, the better the resolution.

Even when a microscope has high resolution, it can be difficult to distinguish small structures in many specimens because microorganisms are relatively transparent. It is often necessary to increase contrast to detect different structures in a specimen. Various types of microscopes use different features of light or electrons to increase contrast—visible differences between the parts of a specimen (see Instruments of Microscopy ). Additionally, dyes that bind to some structures but not others can be used to improve the contrast between images of relatively transparent objects (see Staining Microscopic Specimens ).

  • Explain the difference between magnification and resolution.
  • Explain the difference between resolution and contrast.
  • Name two factors that affect resolution.

Key concepts and summary

  • Light waves interacting with materials may be reflected , absorbed , or transmitted , depending on the properties of the material.
  • Light waves can interact with each other ( interference ) or be distorted by interactions with small objects or openings ( diffraction ).
  • Refraction occurs when light waves change speed and direction as they pass from one medium to another. Differences in the refraction indices of two materials determine the magnitude of directional changes when light passes from one to the other.
  • A lens is a medium with a curved surface that refracts and focuses light to produce an image.
  • Visible light is part of the electromagnetic spectrum ; light waves of different frequencies and wavelengths are distinguished as colors by the human eye.
  • A prism can separate the colors of white light ( dispersion ) because different frequencies of light have different refractive indices for a given material.
  • Fluorescent dyes and phosphorescent materials can effectively transform nonvisible electromagnetic radiation into visible light.
  • The power of a microscope can be described in terms of its magnification and resolution .
  • Resolution can be increased by shortening wavelength, increasing the numerical aperture of the lens, or using stains that enhance contrast.

Fill in the blank

When you see light bend as it moves from air into water, you are observing _________.


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Short answer

Explain how a prism separates white light into different colors.

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Questions & Answers

reactions if viruses to physical and chemical agents?
All viruses contain the following two components: 1) a nucleic acid genome and 2) a protein capsid that covers the genome. Together this is called the nucleocapsid. In addition, many animal viruses contain a 3) lipid envelope. The entire intact virus is called the virion.
assume you have performed a gram stain on a sample of pus from a patients urethra. you see red, nucleated cells and purple rods. what can you conclude
Petty Reply
Study of micro organisation which we cannot see with naked eye...
shyam Reply
Despite their bad reputation, microbes are mostly beneficial or have a neutral effect on our lives. Microbiology is the scientific study of these microorganisms. Microorganisms are those organisms that are too small to see with the naked eye and include things like bacteria, fungi, and viruses.
not 'things'...'organisms' like bacterias, viruses and fungi
simi, it's not bacterias but bacteria which is the plural form of the word while bacterium is the singular of it
bacterium is obvio in a singular form but plural of bacterium is bacterias
Richard is right it's bacterium which is the singular form and plural is bacteria. *Bacterias* doesn't exist.
what is microbiology?
Sandeep Reply
what is microbiology
is the study of small microorganisms that can't be viewed by our naked eyes under a microscope
it's the study of microbes their classification, diversity, their cellular structure and processes, their applications and interaction with other organisms.
what is EMP pathway
which organism is implicated in meningoencephalitis
Peter Reply
Herpes virus
which mold is called dairy mold?
Geotrichum candidum
which infectious agent is the smallest
spore or virus
prion ....it is a single protein in a misfolded form..there is also a theory based on it
yup...m sry
yes it can be turned as a virus
which gram positive bacteria is responsible for food poisoning?
prion is not a living organism yet it is infectoois why
prions are virus like organism. and it is a misfolded protein that may cause disease to us.
Clostridium perfringens, B.cereus
Who is the father of the biology
I have one more question. . that who is the father of botany and medicine? ? can any one tell
define polar bond in simple way
Rashmi Reply
when there is formation of positive and negative poles i.e. one atom is +ve and other is -ve which further results in the dipole moment. Such bond is know as polar bond. It is a convalent bond only.
Hi everyone, is these site only about microbiology? I want a site which I can download articles on hematological and biochemical profile
Peter Reply
I think it is only about micro biology.
so whats happening
I am about to enter school and am going for MCB
Olugbenga Reply
What is DNA damage and repair
Sree Reply
removal of nucleotide bases from DNA is damage n addition or insertion of bases in DNA is repair.
replication is a term related to generation of new DNA from a parental DNA.
role of microorganism as pathogens
Parteek Reply
to live inside the host body, reproduction n destroy the immune system of host body.
what is replication
please give me ans
replication is a term related to generation of new DNA from parental DNA.
difference between endotoxin and exotoxin
Binkheir Reply
toxins released inside the cell or a body is endotoxin n toxins released outside the cell or a body is exotoxin.
Generally, exotoxins are produced by gram positive bacteria and endotoxins are the integral part of the cell wall of gram negative bacteria. But some gram negative bacteria can also produced exotoxins such as E.coli.
Yeah u r ryt aakarti exotoxins screted by gram postive bacteria nd secreted by cell surface and they are secondary metabolites ....On the other hand endotoxin released by gram negtive bacteria nd released inside the cell (lipopolysaccharide) ..
Heat stable lipopolysaccharides to be exact.
please I want the names of bacteria and the diseases they cause.
Ibrahim Reply
there are many we classify it according to their shapes gram negative or gram positive
so u may refer the book u guess or by when u want I can do a list for you and send u here
please do a list for me
I have the lost but it's too much to type ..
MSc entrance prepare books and questions LA etha base pani irukum..
Guhan Reply
kushal always prescott is the imp book to follow.search some objective mcqs books based on your syllabus
deepthi Reply
I want to clear entrance of ms university so I have no more idea so my preparation is based on my bachlor studies.
what are the opportunistic infection in aids stage
certain cancer or pneumonia 🤔
penicillium crysogenum
Kushal Reply

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Source:  OpenStax, Microbiology. OpenStax CNX. Nov 01, 2016 Download for free at http://cnx.org/content/col12087/1.4
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