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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

Can you give an essay of theta model of DNA replication?
ARYA Reply
Actually Bacterial replication is bidirectional process so when replication start in bacterial DNA two dna forks get formed and both replication again meet on same point in termination position ... so therefore in bacterial dna a greek letter (θ) structure formed which is called theta modal.
Bharat
what are thermophile's bacteria
evon Reply
bacteria which can survive in high temperature.
Jayeeta
Thanks
Prudence
why do we study microbiology?
Joyce Reply
For the classification, identification of pathogenic microbes ...
Bharat
thanks
Joyce
what is pathogenicity
jesca
diseases causing activity ...
Bharat
of microbes..
Bharat
sorry ability not activity..
Bharat
I think it's the degree of a pathogen to cause infection
Foster
what is prokaryotes
PRALHAD Reply
are micro-organisms that have no nucleus e.g bacteria
Foster
and they do not possess membrane bound organelles e.g mitochondria
Bharat
yes
Foster
they are single cellular organisms,
Rajasekhar
all the organelles are embedded in cytoplasm of the cell
Rajasekhar
They don't possess true nucleus
PREMI
they're unicellular organisms
Kamaluddeen
they are classified as bacteria and. archaea. bacteria have a cell membrane conposed of peptidoglycan and archaea is composed of psuedomu
Tara
pseudomurein
Tara
organelles are non membraneous as the nucleus
Tara
What is the biosynthesis of a bacteria cell
Adesina Reply
biosynthesis is multiple step enzymes catalyze process which occur in bacterial cell
Jayeeta
biological way of manufacturing food and utilizes it for growth, multiplication and development
Kamaluddeen
prokaryot
PRALHAD
what is microbiology
Kiirya Reply
which organism can not be seen necked eyes is knowm as microbiology
meet
microorganisms
Jayeeta
micro can be define as d smallest living organisms dat can not be see wit naked eyes only with the aid of a microscope 🔬
muhammed
bio means life logy means study
muhammed
biology can b define as the systematic study of living things
muhammed
microbiology can be define as the study of tiny organisms, that's according to my understanding
muhammed
Microbiology is the study of living organisms which cannot be seen by the naked eye but only with the aid of a microscope. These microorganisms can be classified as bacteria, fungi, micro-algae, protozoa and virus.
Tatz
microbes that can't seen in naked eyes. upto 100micrometer we can see in naked eyes but 1 microns or nanometers are invisible in naked eye.
Jayeeta
what is a cell
Oteri Reply
cell can be define as smallest indivisible organism in the body
muhammed
cell is the smallest unit which makes up the whole body
Yue
cell is smallest unit and there are two types prokaryotic cell and eukaryote cell. bacteria a prokaryotic cell which works independently.
Jayeeta
nice forum
muhammed
Which parasite competes with the host for vitamin B12 to cause pernicious anaemia
Abubakar
Diphyllobothrium latum for vit B12
Jayeeta
it is also fish tapeworm
Jayeeta
Diphyllobothrium sp feed vitB 12
Jayeeta
recent trends in microbiology of biofertilizer
Ykomalipriya Reply
using nano technology
Jayeeta
what is transposors
EMMANUEL Reply
Junk DNA, jumping gene.
Pradeep
transposon elements are jumping gene as DNA sequence move from one location to anothet
Jayeeta
Yes
Pradeep
what is microbiology
Usama Reply
Microbiology is the study of organisms, most of which are too small to be seen with the naked eye, including bacteria, algae, protozoa, and fungi, as well as acellular agents, such as viruses and prions.
digvijaya
what is gram +ve and gram - ve bacteria?
Sharma
gm- contain lipid layer so it take counter stain during gram staining and pink or red in color but gm + does have thick LPS layer.
Jayeeta
what year microbiology started
Kartik
Robert Koch was the first to detect microorganisms
Jayeeta
what appears microorganisms
Kartik
clear the questions please
Jayeeta
was it robert koch or robert hooke?
digvijaya
microbiology word came from microns. study microbes that not recognize in naked eyes.
Jayeeta
what are the branch of microbiology
Oyemade Reply
We can basically classify into pure and applied microbiology
Ozzie
what is arabinose operon and it's stucture
Pooja Reply
The L-arabinose operon, also called the ara or araBAD operon, is an operon required for the breakdown of the five-carbon sugar, L-arabinose, in Escherichia coli
Tharra
kya hum operon ko divide kar skate hain
Kartik
reactions if viruses to physical and chemical agents?
VICTOR Reply
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.
Asali
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
bacteria
ABUBAKAR
gram negative bacteria hence gonococcal infection
Kiirya
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.
Asali
yes
Dhirender
not 'things'...'organisms' like bacterias, viruses and fungi
Sinmi
simi, it's not bacterias but bacteria which is the plural form of the word while bacterium is the singular of it
Richard
bacterium is obvio in a singular form but plural of bacterium is bacterias
Sinmi
Richard is right it's bacterium which is the singular form and plural is bacteria. *Bacterias* doesn't exist.
Aakriti
that is microbiology which is deals with microbes only their structure
Kiirya

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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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