Defined : the emission of light as a result of a chemical
reaction during which chemical energy is converted into light
energy From a living organism that functions for its survival or
mating Comes from Greek word bios for living and Latin word lumen
for light, so it literally means living light Cold light resulting
from a specific biochemical mechanism involving chemical processes
Specific for that organism Found all over biosphere,
phosphorescence in sea water observed in all oceans Bioluminescence
of visible light is found in a majority of marine organisms but is
rare in terrestrial organism
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Slide 5
All involve an oxygen oxidation of organic molecule (luciferin)
Catalyzed by an enzyme called luciferase These proteins are called
photoproteins -> oxygen is already bound to the luciferin A
photoprotein is a protein with a luciferin bound to it Antenna
proteins -> adjust the colour of bioluminescence likeness to
proteins on similar function in photosynthesis except they act in
reverse 5 known distinct chemical classes of luciferins: Aldehydes,
benzothiazoles, imidazolopyrazines, tetrapyrroles and flavins Key
organ -> photophore (light producing organ) Seen in many
luminous fish and vividly in cephalopods Make up of complex
photogenic (light emitting) cells Bioluminescent reaction
components detected in stomach, secretory organs and liver of some
organisms
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Bioluminescence: light is created through a chemical reaction.
Bioluminescence is a subset of chemiluminescence Fluorescence: a
particular atom or molecule absorbs light of one length, but emits
one of a longer length. The light going in is usually UV photons,
and comes out as visible light Phosphorescence: requires more time
to remit radiation absorbed than fluorescence. This is because the
sub atomic reactions required to remit light occur less often than
in fluorescence. Fluorescence
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Stories of mysterious light or fires seen over fields or
mountains were often said to be dragons or the gods Greeks and
romans first to report luminous organisms Reports of fireflies
found in early religious writing of India and China Earlier
recordings believed to have come from these and ancient eastern
civilizations and refered to firelies and glow worms Aristotle
(384-322BC) (first to discover) decribed more than 180 marine
species, first to recognize cold light Later, complete and
extensive descriptions of luminous organisms published by Pliny the
Elder (23-79 CE) 16 th century references to bioluminescence found
in literature such as Shakespeare, specifically in hamlet who
talked about effectual fire of the glow worm First book devoted to
bioluminescence and chemiluminescence published in 1555 by Conrad
Gesner 1667 Robert Boyle documented oxygen was needed for
luminescence
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Raphael Dubois performed experiment, he extracted the two key
components of bioluminescence reaction and was able to create light
and that there 2 key components= luciferine and heat labile
luciferase* One of most eminent scientists of 20 th century was
Princeton Professor E. Newton Harvey* He was looking for existence
of lucifernin- luciferase system in all luminous organisms First
luciferin isolated in 1956 First photo protein isolated was the
calcium activated photoprotien aequorin in the 1960s
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Calcium dependant photoprotein cloned in 1985 Intensity of
luminescence varies with calcium concentration Therefore aequorin
has been used in monitoring of cell calcium 1985 firefly luciferase
was cloned Firefly Tomopteris Luciferin chemical structure
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All colours of the visible light spectrum; red, orange, yellow,
green, blue, indigo, violet Different colours dependant on role the
light plays and which organism it is produced in Many only produce
one colour, however some are capable of many colours Eg. Jamaican
click beetle; this is due to the same luciferin substrate as the
firefly, but different luciferase structures Bioluminescence
releases a large amount of energy, not heat Visible light radiation
is equal to light wavelengths of 400- 700 nm Bioluminescence max of
most marine species is 460-510 nm Terrestrial organisms mostly
yellow-green BL, more yellow Marine mostly blue-green (400-500nm)
luminescence because it travels the best through water
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Plays an important role in nature due to the darkness 200m
underwater Location on body of bioluminescence gives clues to the
functional role of the luminescence Attracting a mate Eg. The wave
lengths attract mates, but do not draw attention to themselves
Attracting prey Eg. Angler fish has a luminescent lure Finding food
eg. Loose jaw fish, bioluminescent organs in cheeks to see in dark
water Communication eg. Fireflies flash certain patterns to either
attract mates or communicate Camouflage eg. Bobtail squid blends
into background Defense against predators eg. Deep sea shrip vomits
bioluminescent material in direction of attacker Deep- sea shrimp
vomits bioluminescent material
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R OBERT B OYLE British philosopher and scientist In 1667
performed experiment and found that bioluminescence didnt occur
when a known bioluminescent type of fungus was not in the presence
of air 1672 he discovers that bioluminescence reactions require
air, when he later discovered oxygen, we realize is the oxygen
component of air that is needed Robert BoyleOxygen Atom
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R APHAEL D UBOIS 1887 he discovered luciferin and luciferase
through experiments with clams, beetles, and other species He did
an experiment using a clam called the common paddock Ground tissue
of the clam up in cold water, and light was produced for several
minutes. This showed he has extracted the light producing chemical
Then made a hot water extract from another clam and added it to the
cold water, which reactivated the light reaction A hot water
extraction alone produced no light reaction Called the hot water
extraction luciferin Called cold water extraction luciferase Noted
that luciferin could only glow in the presence of luciferase
Raphael Dubois
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E DMUND N EWTON H ARVEY Greatly popularized the study of
bioluminescence Discovered that luciferins and luciferase from
different animals are not interchangeable Evidence of evolution of
bioluminescence to fit various needs of different species Now shown
that evolution of bioluminescent systems has occurred over 30
times, which accounts for the differences in colours and uses in
different species Humpback Angler Fish Jamaican Click Beetle
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The discovery and study on bioluminescent organisms is the goal
of many expeditions of ocean going research vessels and marine
submersibles C OMMERCIAL A PPLICATIONS Glowing trees to save
electricity bills Agricultural crops that luminesce when in need of
water Detection of bacteria in contaminated foods Novelty pets
Bio-indentifies for convicts, mentally ill Glowing toys, glowing
greeting cards Luminescent beer and champange Glofish
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M EDICAL A PPLICATIONS Reporter genes Bioluminescent Imaging*
In vivo analysis Detection of bacteria Observations of protein to
protein interacting Testing for genetically modified organisms*
Water quality testing* Green Florescent Protein*
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Used on small animals Real time monitoring of the progression
of infections in the same animal Typically 2D imagery, with lower
resolution Can mark progress at different time points without
euthanizing the animal, uses animals mice than conventional methods
of progress tracking Quicker and relatively inexpensive Allowed
specific molecular and cellular events such as cell migration and
signal transduction to be investigated in a living, intact
animal
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Bioluminescence is decreased by pigmentation of organs such and
liver, spleen and the fur To get around this they can shave the
mice or use ones with the albino gene Used to study bacteria
distribution, distinguish between more and less virulent strains
and moniter antibiotic therapy Also used to study viruses, but
because of low resolution, it can be difficult to distinguish
between viral infections in adjacent tissues Used to investigate
parasite infections High correlation between light intensity and
amount of parasites in spleen Mouse and BLI
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For water quality/ toxicity testing Uses the bioluminescent
marine bacteria vibrio fischeri When the organism is challenged by
a toxin, its breathing pathway is disrupted, resulted in decreased
bioluminescence intensity Now a well established and excelled
genetag and protein It can be fused to a protein of interest and
fluorescence and can be tracked within a cell to study its
localization and behavior Outstanding structural stability
Excellent for studying the cell and sub cellular processes Cloned
in 1992, expressed in various organism in 1994 Vibrio Fischeri
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People want to know, Does this food contain genetically
modified organisms? and they have the right to know Genetically
Modified Organisms (GMO): genetic material (DNA or RNA) that has
been altered in ways that would not occur under natural conditions
or natural processes Most common technique used to test for GMOS is
polymerase chain reaction (PCR) PCR requires complex DNA extraction
techniques, rapid thermocycling, expensive equipment, and is a
lengthy process Company called Lumora came up with a breakthrough
technology to test for GMO Its a combination of 2 technologies;
Bioluminescence and isothermal DNA amplification Bioluminescence
used is bioluminescence real time reporter (BART) Organic food
symbol
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Loop mediated isothermal amplification (LAMP) BART uses
luciferase and detects DNA and It lights up when it find specific
DNA and RNA sequences linked to genetic modifications GMO testing
can be done out in the field or in a food processing center
LAMP-BART technique requires only basic equipment for DNA
extraction, a constant temperature and simple light detection
Quicker than PCR Good for farmers to have a cheap, reliable way to
test products for GMOs Lumoras GMO detector can also be used to
test for things like salmonella in foods
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What are the biological advantage of light emission to the
animal? Evolution history? Metabolic/ dietary source of the
luciferins? What are the control mechanisms for light flashing?
Discovering- luminous mollusc, a roman delicacy, the
bioluminescence mechanism of it is still not completely solved
Slide 23
Deep-sea Shrimp
http://www.nytimes.com/imagepages/2011/12/20/science/20JPBIO1_SPAN.html
Chemical equations and fluorescence animations:
http://www.lifesci.ucsb.edu/~biolum/chem/ Tomopteris
http://discovermagazine.com/photos/1-8-marine-creatures-that-light-up-the-sea
Chemical Structure drawing of Luciferin
http://www.biotium.com/product/applications/Enzyme_substrates/price_and_info.asp?item=10100&layer1=D;&layer2=D01http://www.biotium.com/product/applications/Enzyme_substrates/price_and_info.asp?item=10100&layer1=D;&layer2=D01;
Robert Boyle
http://www.friedpost.com/sciencetech/greatest-science-discoveris-boyle%E2%80%99s-law-1977.html
Raphael Dubois
http://en.wikipedia.org/wiki/File:Raphael_Dubois_(1849-1929).jpg
Oxygen Atom
http://www.historyforkids.org/scienceforkids/chemistry/atoms/oxygen.htm
Bioluminescent Imaging mouse
http://www.caliperls.com/products/preclinical-imaging/?gclid=CMiigb2T_q8CFWwDQAodmVZCzg
Glofish
http://fish-blog.co.uk/News/manufacturer_supports_glofish_industry/
Vibrio Fischeri
http://microbewiki.kenyon.edu/index.php/Vibrio_fischeri
Bioluminescent Imaging mouse
http://www.caliperls.com/products/preclinical-imaging/?gclid=CMiigb2T_q8CFWwDQAodmVZCzg
Organic food stamp
http://www.google.ca/imgres?um=1&hl=en&sa=N&biw=1366&bih=569&tbm=isch&tbnid=pcYC93zfdGquhM:&imgrefurl=http://www.omafra.gov.on.ca/e
nglish/crops/organic/certification.htm&docid=MuinqDkP2Ld1-
M&imgurl=http://www.omafra.gov.on.ca/english/crops/organic/certificationf10.jpg&w=600&h=616&ei=JSGwT6XnFarE6QG1rqCvCQ&zoom=1
Humpback Angler Fish
http://written-4u.blogspot.ca/2007/01/allah-creator-sustainer-surely-it-is.html
Jamaican Click Beetle
http://www.asknature.org/media/image/6970
Slide 24
Beecher, C. (2012, May 7). Breakthrough Offers Promise of
Improved GMO Testing. In Food Safety News. Retrieved May 9, 2012,
from
http://www.foodsafetynews.com/2012/05/breakthrough-offers-promise-of-improved-
gmo-testing/ Binger, J. M. (2007, March 14). Bioluminescence. In
Center for Biophotonics Science and Technology. Retrieved May 6,
2012, from
cbst.ucdavis.edu/education/courses/spring-2007./bingerfinaldraft.doc
Hutchens, M., & Luker, G. D. (2007, June 24). Applications of
bioluminescence imaging to the study of infectious diseases.
Retrieved May 7, 2012, from Wiley Online Library
(10.1111/j.1462-5822.2007.00995.x). John, L. (08). Basic
Bioluminescence. In Photobiological Sciences Online. Retrieved May
7, 2012, from http://www.photobiology.info/LeeBasicBiolum.html
http://www.photobiology.info/LeeBasicBiolum.html Lee, J. (08). A
History of Bioluminescence. In Photobiological Sciences Online.
Retrieved May 6, 2012, from
http://www.photobiology.info/HistBiolum.html
http://www.photobiology.info/HistBiolum.html Lumora welcomes
expressions of interest for GMO test commercialisation after BART
technology demonstrates ability to detect genetically modified
contamination of crops rapidly and at very low levels o. (2012,
April 30). In Lumora. Retrieved May 9, 2012, from
http://79.170.44.87/lumora.co.uk/index.php?option=com_content&view=article&id=100:lumora-welcomes-
expressions-of-interest-for-gmo-test-commercialisation-after-bart-technology-demonstrates-ability-to-d
http://79.170.44.87/lumora.co.uk/index.php?option=com_content&view=article&id=100:lumora-welcomes-
expressions-of-interest-for-gmo-test-commercialisation-after-bart-technology-demonstrates-ability-to-d