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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

microbiology is the study of micro organisms those been unicellular multicellular or acelluar
Nwakaego Reply
what is microbiology
Ebuka Reply
why study microbiology
David Reply
to know the cause of certain disease and how to cure them
Racheal
without micro, medicines in term of diagnostic and cure would be far away from realizing its objectives
Louisiaste
It's help to give the best medication.
Olayinka
Unvin Prisca the immune system
Gyabaa Reply
immune system is a complex of organ tissues and specialized cells which identify or kill the pathogens.
Mohammad
Thanks
shamim
explain to me the pathogenesis of plasmodium
Angelo
what is difference between bacillus and ecoil
Rymes Reply
similarities between prokaryotic cell and eukaryotic cell
Ednah Reply
Eukaryotic cells contain membrane-bound organelles, including a nucleus. ... Prokaryotic cells do not contain a nucleus or any other membrane-bound organelle. Prokaryotes include two groups: bacteria and another group called archaea.
Priya
thanks
Sarpong
what is different between plasma and serum?
Alif
difference between plasma and serum is that plasma is liquid n serum is fluid
Priya
what is microbiology
Eliham
what is the diseases caused by the microorganisms
Emmanuel
both cells have plasma membrane, which services as a barrier between the inside of the cell and outside. both prokaryotes and eukaryotes have ribosomes. they receive messages from messanger RNA molecules, telling them what proteins the cell needs.
Abdinur
Thanks
Dominic
Some bacteria infection
Dominic
what is the name given to infection got from the hospital
Mukisa
Nosocomial infection
Gifty
what is the apportion
Riyaaq
what is the main functions of organelles
Ishaq
what is main function of right and left side of the brain.
Theophile
Salifu. microbiology is the study of oganisms that are too small for the naked eyes to see. this include bacteria
Theophile
Cabdi. apportion is to divide or share out
Theophile
yap
bako
what is the parasitic helminths?
bako
what are pathogens
Oluka Reply
any disease causing microorganism that can invade the body and cause harm
famuyiwa
hepatitis b
Oluka
what's da answer
Oluka
any diseases that course harmful to body
Musa
hepatitis is usually implementery of the liver and is commonly course by virus infection
Musa
a type of virus that causes severe illness lasting from two to six weeks it may cause massive lived necrosis and cirrhosis
famuyiwa
hepatitis B are transmitted when in contact with body fluid such as blood,semen,varginal secration that contain the hepatitis b virus
Musa
I understand necrosis,means death of all cell or injury of blood supply, but what is cirrhosis pls
Musa
sorry meant liver cirrhosis
famuyiwa
good evening all
Billy
ok liver damage
Musa
I have a question
Billy
Hey welcome all of you...i think you are discussing about Micro-organisms..what does it Micro?
Gazu
how about, scabies (bacteria) yeah can some one gist me abt it
Billy
sorry what doest it mean Micro organism?
Gazu
microorganism are minute living organisms found almost everywhere. they can be autotrophic heterotrophic or chemotrophic
famuyiwa
Tell me about enzyme kinetics
Jaypal
wowoww thanks a lot farmuyiwa Alice
Gazu
have anyone colony pigmentation of different types of bacteria such as Aeromonas, Edwardsiella, vibros,
Sadiqur
yes dear sadiqur Bacterias are live in everywhere.. .
Gazu
microorganisms are arganism that cannot been seen with 👀 until by the use of microscope
Musa
microorganisms are arganism that cannot been seen with 👀 until by the use of microscope
Musa
yhea organisms are organisms they are naked eye/can't seen by our eyes
Gazu
and scabies are one of the most common skin condition that can course itches and raches.
Musa
and also scabies are not infection, but an infestation
Musa
my Q is what is the contribution of van-leeuwenhoek in field of microbiology
Musa
Antony van leevenhoek is father of microbiology.
Jaypal
Anton Van leewun hook is discovered Microscope it's true or not? who can brief me please
Gazu
micro organisms are organism that cannot be seen with an aided eyes ,but by the use of microscope
Ssmuel
Yes.. He discover first microscope. He take 2 lence at a time and he saw the plant cells... Thus he was called father of microbiology
Jaypal
who examined water from a bathtub and discovered microbes
famuyiwa
for instance any type of Stagnant water is creat there are d/t tupes of small animals.. so those animals are called Micro organism?
Gazu
thank you javpal for Describing.
Gazu
Hi the father of microbiology is Louis Pasteur
bako
Then the father of medical microbiology is Robert Koch
bako
can you please differentiate between vacuoles and inclusion bodies
Noor Reply
vacuoles consists of watery solution called cell sap while bodies are organelles
Prince
5 difference please
Noor
Archaea is an odd type of bacteria that live in extreme environments; high salt, high temperature etc. usually called extremophiles.
Danesia Reply
you know anyone that has delt with enterobacter agglomerans.
Vernon
it is a type of gram negative bacteria and its ubiquitous mostly isolated from plant stuffs and also from faecal material of animals. I heard the name pantoea . it is bacillus in structure.
Sujan
OLUKA MARTIN
Oluka
What are the classification of microorganisms
Portia Reply
Basic classification are eukaryotic and prokaryotic..
micro organisms are classified into three namely: Archaea, eukaryote and prokaryotic organisms
Blandine
Yay that's right
what is archaea
Ateeq
I've got an issue with the message view
Frank Reply
Is there any effective way to learn microbiology.
Frank Reply
I have some basic knowledge of biology. I mean i'm totally beginner in that field.
Frank
Assuming that i'm interested in neuroscience. What is the best way to master that particular subject.
Frank
Hi
Can you wait me just a minute
Iam apologies for guidance you in neuroscience.. I'm not physician.. I'm microbiologist
My pleasure to join to you
Let's begin a conversation
Frank Reply
What would be saying
?
Akram
do you know anything about microbiology
diana
I have so much studying to catch up on🙄
Britnee
any study tips
Britnee
Yas of course
What would you like to know
well i'm studying Pharmacy second year and i just started With Microbiology
diana
if you have any basics to tell us 🤷🏻‍♀️ would be helpful
diana
Yas, first you must know that you will have a very easy lecture..
In second stage you will try to learn the type of microbiology
A simple definition for microbiology
In second stage you will try to learn the type of microbiology.. you must learn the bacteria..its structure.. Classification of bacteria according to shape..
It's so easy
What triggers the natural killer cells
jenipher Reply
the immune system
Unvin

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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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