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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

give me the name of good microbiology books to download
Shahid Reply
Who invented microbes?
manor Reply
Which year did Robert koch invented the microbes?
manor
Robert Koch
Matilda
Antony Van Leeuwenhoek invented microbes through a designed single lensed microscope.
Mohamed
what is microbiology
Nyoik Reply
microbiology is the study of micro organisms. this can be multicellular,unicellular & acellular
Janet
what is the difference between DNA/RNA
kanaaLka
DNA is a double stranded molecule while RNA is a single stranded molecule
Lovely
who is the inventor of microscope
Alberta
What are antibodies?
Happy Reply
they are immune cells that are released by immune system to fight strangers like microbes
Ahmed
a blood protien produce in response to an counteracting a specific antigen
Pooja
Why salmonella typhi is harmful protozoan?
Saily Reply
because make typhoid that is considered as a food born illness
Ahmed
Name any five modes of transmission.
Happy
what are the scientific names and common names of some microbes
Rich Reply
Staphylococcus aureus - staph Streptococcus pyogenes - strep Botulism - Clostridium botulinum Rocky Mountain spotted fever - Rickettsia rickettsii Diphtheria - Corynebacterium diphtheriae Gonorrhea - Neisseria gonorrhoeae
Tammy
what are the limitations of the Koch's postulate
Naa
how a microorganism growth.
Zubair Reply
Is foetus a parasite to his or her mother? if yes and if no ho
Mashauri Reply
y
Yusri
yes
Yashkin
yes
Zubair
yes it a parasite to the mother because it feeds on the mother for survival
Beatrice
yes
Redwan
no
Elasha
definition of a parasite: an organism that lives in or on an organism of ANOTHER species (its host) and benefits by deriving nutrients at the other's expense
Elasha
no because an organism can inky be a parasite if it causes harm to its HOST. And the fetus does not cause harm to its mother under normal conditions
aliyu
yes
Naa
what is biosensor in microbiology
Raja Reply
What are biosensor
Raja
what's are biosensor
Raja
what is microbiology defination
Rinku Reply
microbiology is the study of small or manuit organisms which cannot be seen with our nacked eyes unless with the aid of the microscope
Brandina
is the study of living organisms which are not directly visible to a direct eye but can only be seen under a microscope
Ipa
ok
Ipa
microbiology is the study of living organisms of microscopic size it is also the study of micro organisms with their form structure reproduction psychology metabolism and classification
Priyanka
ok
Aminu
Microbiology are the study of microorganisms either microscopic or sub microscopic creaters mainly unicellulars, multicellulars and subcellulars. Such as protozoa,bacteria and viruses.
Rana
it is the biological study of viruses, fungi, protozoa, bacteria which in collective name are called micro organisms, unlike microscopic organisms being invisible, that requires a magnifications with the help of a microscope.
Mohamed
what are the importances of Microbiology?
Erick Reply
don't know
SINGLE
we obtained the insulin from the bacteria. and some microorganisms are decomposer in ecosystems.
Zubair
nitrogen in the air is fixed into the soil by microbes example is nitrobactor
Matilda
yes
Zubair
how a weak immune region where microorganisms attack easily?
Zubair Reply
with examples differentiate gram positive from gram negative bacteria
Mary Reply
Differentiate gram positive from gram negative
Mary
I have no idea
Zubair
hello
Kuonain
and example of gram negative is E. coli
Pooja
gram positive stain purple when subjected to gram stain whilst gram positive bacterial has thick wall composed of peptidoglycan
Matilda
ok
Zubair
what is a process of gene expression in eukaryotes ?
Nayyab Reply
pls help us with the answer
Abdussalam
spontaneous generation means
Siddhi Reply
hiii
Siddhi
Hello
eman
So, what's next 😂
eman
I have a problem with Micro
eman
I studying things and see question Some thing else
eman
Can u help me, please
eman
What i havta do
eman
firsfall when you studying understand things...
Siddhi
good afternoon dear Friends
yaya
Greeting to everyone in here.
Manka
There is a problem. I need a diagram of a virus with it functions.
Manka
structure of bacterial
Kuyiba
gghhhh
Zubair
group plz i need help in microbio
Kuonain
it really difficult fr me
Kuonain
kashur cha kah
suhail
sorry now I read in class 8th but I can help u
Zubair
Me too. It's very difficult for me
Angela
rod shape
Priyanka
helical
Priyanka
Hello
esike
spherical
Priyanka
peomorphic
Priyanka
I am ryt or not
Priyanka
hi
suhail
hello
DIPTI
Are we together pls what's the topic for the day?
esike
what is innate
Lizzy
innate means natural
Pooja
the genetic makeup of an individual
Matilda
pls i need the common names for the following parasites..ENTEROBIUS VERMICULARIS,NECATOR AMERICANUS,ASCARIES LUMBRICOIDES,TRICHURIS TRICHIURA,TRICHOMONAS VAGINALIS,GIARDIA LAMBLIA,ENTAMOEBA HISTOLYTICA,SCHISTOSOMA MANSONI,SCHISTOSOMAHEMATOBIUM,STROGYLOIDES STERCORALIS,AND TRAPANOSOMA BRUCI GAMBIENSE
timothy
ENTEROBIUS VERMICULARIS and STROGYLOIDES STERCORALIS have the same common name which is pin worm and thread worm
Chinedu
I didn't find the common names for TRICHOMONAS VAGINALIS, GIARDIA LAMBLIA, ENTAMOEBA HISTOLYTICA, TRYPANOSOMA BRUCEI GAMBIENSE
Chinedu
NECATOR AMERICANUS - New world hookworm ASCARIS LUMBRICOIDES- Ascarid TRICHURIS TRICHIURA- Whip worm
Chinedu
Hi guys
Chinedu
Why are myeloblasts not present in the bloodstream?
Chinedu
thank u
Dasaah

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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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