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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

how do i report widal slide results
Isaac Reply
how food can be used as a substrate for the growth of microorganisms
Ashi Reply
what is the importance of understanding chemistry in the field of microbiology and food technology?
Charmaine Reply
What is different between eukaryotes and prokcaryotic
Abia Reply
main difference is that eukaryotic cells possess membrane bound organelles
Iqra
prokaryotes are primitive organisms that doesn't contain membrane bound nucleus or any orgenelle while in eukaryotes membrane bound orgenelles and nucleus is present
Yashi
what s anatomy
jane Reply
its mean body structure, function, and systems
gazi
study of internal structure of living things
Falere
anatomy is the study structure of the made human
isir
anotomy is the actual study of body internally and externally Which include how itis made. for what and what is need of this
Iqra
la science anatomie c'est la science qui nous aide à étudier l'homme
Baraka
okay
Baraka
Ancestor are they real
Rapheal Reply
yes of course
David
yes please
Clark
50 50, depending on the accuracy of the clan records.
Vincent
okay
Baraka
Why protist is not a kingdom of Linnaeus 'S taxonomy?
Neha Reply
I dont know
Jeewraj
Linnaeus used Aristotle's criteria for dividing living organisms into kingdoms. Plants are immobile and insensitive, while animals are mobile and sensitive. the creatures he saw through the microscope were mobile, so he attributed them to animals.
Lad
name the different types of media use in lab to detect the micro organisms
Shehzadi Reply
The different types of media used in the lab to detect the microorganisms is known as cell culture..
Tean
solid media or broth is used to grow and detect microorganisms
Pohor
what is microbiology
Baraka
the study of large living organisms
Pratibha
the study of organisms which are micro in range
Vency
what is autoclaving?
Yashi
process for sterilization
Vency
is a machine used on the process of sterilisation
Clark
what is difference between hot air oven and autoclave as they both are used for sterilization ?
Vency
autoclave basically do moist heat sterilization while hot air oven do sterilization by dry heat.....
Yashi
what is the microbe
HUSSAIN
which method out of these two is best?
Yashi
why human have microbe
HUSSAIN
Hi
Sadam
hi to all
Sadam
depends on what to sterilize
Vency
microbes
Md
what is knowledge
HUSSAIN
well idea
Daniel
what are the importance of microbiologe to a nurse
Enny Reply
it enable a nurse to know the weight and height of a patient
Abotu
it helps the nurse in way as to give first aid to patient on which basis doctor will take diagonos
Iqra
disagree with both of you
Akhtar
ok
Abotu
Your Own idea
Abotu
it helps a nurse to be able to counsel a patient /client
Abotu
cell culture
Tean
Types of micro organisms
Jennifer Reply
Who discovered plant and animal cell
Jennifer
Robert Hooke
Martha
Father of microbiology
Jennifer Reply
Antonie van Leeuwenhoek
Satarupa
Why are mitochondria and chloroplasts unable to multiply outside of a host cell?
Iqra Reply
iqra mitochondria arises from the division of existing mitochondria and they are fused together. they move around inside the cell with the interactions of the cytoskeleton that's why mitochondria unable to multiply outside the host cells chloroplasts has its own separate DNA from the plant cell.
Rana
short not on medical microbiology
Massah Reply
Hy
Iqra
hope you fine
Arif
love you
Arif
Assalamualaikum
Suhaib
arai yeh micro kai sth kis ko love hua
Umer
icrobiology 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
Suhaib
iqra iqbal
Arif
What is gram syaining?
Arif
to differentiate between gram positive and gram negative bacty
Umer
True
Arif
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Arif
i m frm sopore..kashmir
Umer
i m frm baramulla .. Kashmir
Umer
and u
Umer
umer
Arif
and i m from afghanistan lovely country
Arif
yes
Umer
thanks
Arif
medical microbiology means study of microorganisms which are beneficial or harmful for the body. mostly which are studied under microscope by structure and shape appearance. e.g bacteria, viruses, bacteriophage, fungus hyphae and some blood and other body fluids parasites e.g plasmodium etc.
Rana
arif bhai gram staining in which we are given different dyes to the bacterial cell wall and cytoplasm and nucleus.e.g giemsa stain
Rana
Gram staining uses crystal violet to stain cell walls iodine as a mordant and a fuchsin or safranin counterstain to mark all bacteria
Umer
Gram staining uses crystal violet to stain cell walls iodine as a mordant and a fuchsin or safranin counterstain to mark all bacteria
Umer
Gram staining uses crystal violet to stain cell walls iodine as a mordant and a fuchsin or safranin counterstain to mark all bacteria
Umer
Omar baba crystal violet iodine 70% alcohol fuchicine or safranine after every step washing must be done
Rana
hello
Cristy
*Fuchsin stain
Cristy
*Safranin
Cristy
thanks
Ganesh
difference between epidermophyton trichophyton and microsporum
Ankita Reply
plz answer the question
Iqra
epidermophyton type of fungi causes superficial and cutaneous mycoses trichophyton is also fungi type including parasitic varieties causes dermatophytosis microsporum is also type of fungi causes dermatophytosis.
Rana
ok
Iqra
Is this app perfect for preparation of exam?
Ankita
Ankita I totally disagree with your respected opinion. it's just minor helpful
Rana
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Ankita
what is infectious
Fred 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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