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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 _________.

refraction

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Short answer

Explain how a prism separates white light into different colors.

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Questions & Answers

Explain about enzyme transportation
Shahla Reply
Enzyme transportation
Shahla
it looks reallllyyyyy coooooooollll. i love enzymes theyre so cooll and i also like protein transportation so i think it would be really cool so an enzyme transport something so ill do some research and come back to ya in that
maxo
they be ridin' dirty
Bethany
what is the infectious disease process
Patience Reply
what are differences between endotoxins and exotoxins
sabote Reply
endo toxins work in the nuceus. i think
maxo
tell me if im right tho
maxo
Exotoxins are toxic substances secreted by bacteria and released outside the cell. Both gram positive and gram negative bacteria can produce and secrete exotoxins. Whereas Endotoxins are bacterial toxins consisting of lipids that are located within a cell. Only lysed gram negatives.
Abdi
Remebr the Lipid A portion of LPS is what's toxic.
Abdi
oh yeah. thanks
maxo
Your welcome :)
Abdi
How did you learn this?
maxo
For me personally the best book is 'microbiology made ridiculously simple'
Abdi
I got my basics from there and slowly added information from other sources.
Abdi
thats cool! yeah i like microbiology too! especially the molecular proteins theyre sooooooooooo cool!
maxo
what are the prokaryotic
Lungu Reply
prokaraytotic is a unicellular organizm that lacks membrane bound nucleus
Zaajid
and whats eukaryotic
abdiqani
eukaryotic cell are cell which contain anuclues and organells
Zaajid
eukaryotes are the cells that have organells which are protected by membranes
maxo
eukaryotic is are multicellular organisms which are open nucleus.
Serah
Explain on the Francisco reddi did to prove the theory of spontaneous generation
Diana Reply
what is parasite
Abdirizack Reply
parasite are organisms feeds on a host for food and survival.e.g round worm for animal, Dodder for plant parasites.
Serah
parasite are organisms that feeds on a host for food and survival.e.g round worm for animal and mistletoe for plant parasites.
Serah
parasite are organisms that feed on their host
Cylla
designing of aseptic area
Aashish Reply
I don't know
Abdirizack
what is rickettsia
DENNIS Reply
what is microbiology
Erasto
Is the science that works with microorganisms.
Rose
richettsa is small microorganisms that cause disease in human like typhus; they are like viruses that can grow only inside living cells, they're transmitted by mites, ticks or lices.
Rose
what is plasmid?
mavis
plasmid is extra chromosomal body present in bacteria...which have additional genetic functions example... antibiotic resistance genes....etc etc
Chaitanya
state the theory of spontaneous generation of micro oranisms and germ theory of disease
UKAMAKA Reply
what are the advantages of high note numerical aperture
Genius Reply
list if non flagellated pritozoa
Mepung Reply
Can someone that's understanding of the Kreb Cycle please explain & breakdown it down to me in the simplest way without giving me the dictionary version or Google version. Basically in there own words of knowledge....!
Kisha
ok
lucas
please can someone help explain positive and negative feedback in simple term
Gum
negative feedback is the arresting of reaction or reverse of the reaction according to the response and postive feed back is the direct response without reversing or arresting a reaction.
Greet
OK
umar
pls can someone explained Kinney stone in memorising in shot time
umar
what tyoebif microorganism will be killed by antibiotic trwatmeant
Mary Reply
I don't really don't understand aerobic respiration anaerobic respiration and fermentation. Can someone explain & breakdown this in the simplest way please.
Kisha
Anaerobic Respiration in which foodstuffs are partially oxidized, with the release of chemical energy, in a process not involving atmospheric oxygen, such as alcoholic fermentation, in which one of the end products is ethanol.
Muhammad
aerobic respiration A type of respiration in which foodstuffs are completely oxidized to carbon dioxide and water, with the release of chemical energy, in a process requiring atmospheric oxygen.
Muhammad
please can someone explain what pseudomonas species and biofilm is?
uju
Fermentation is the growth of cells or microorganisms in bioreactors (fermenters) to synthesize special products. Fermentation in biochemistry refers to the biodegradation of carbon compounds by cells or organisms under anaerobic (lack of oxygen) conditions.
Muhammad
Please under which conditions does pathogens become established in the human tissues and can cause diseases
Atambilla
please the life cycle of plasmodium parasite
mavis
who is John Needham
Mary Reply
John Needham is one of the researcher in microbiology. He also experimented when scientists did not believe animals could arise spontaneously ,but did believe microbes could.
Gum
okay is he late
Mary
by the way what are the list of courses offered by a newly admitted student for microbiology
Mary
Needham's experiments with beef gravy and infusions of plants material reinforced this idea.
Gum
what drugs are given to a person with Otis nerve problem(ear problem)
Gum Reply
good morning to you all.
Muhammad Reply
Dr.A.K.S.P.G. college Akbarpur Ambedkar Nagar
Rajan
Shailesh Vishwakarma BSc 1st year
Rajan

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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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