Fluorescence:

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Fluorescence an effect in which a substance
releases electromagnetic radiation while
absorbing another form of energy, but ceases to
emit the radiation immediately upon cessation of
the input energy the light emission of a given
wavelength by a substance that is activated by
light of a different wavelength. Academic Press
Dictionary of Science and Technology, 1992. The
emission of a longer wavelength radiation by a
substance as a consequence of absorption of
energy from a shorter wavelength radiation,
continuing only as long as the stimulus is
present distinguished from phosphorescence in
that, in the latter, emission persists for a
perceptible period of time after the stimulus has
been removed. Stedmans Medical Dictionary, 24th
ed., 1982.
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http//chsfpc5.chem.ncsu.edu/Poznan/membrane_fuse/
index.htm
micro.magnet.fsu.edu/.../ jablonski/lightandcolor/
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General Resources http//www.probes.com/handbook/
Fluorescent Probes www.sigmaaldrich.com/.../
Labels.html
Choice of Fluorescent Labels www.aecom.yu.edu/aif
/instructions/ probes.htm
BioRad Fluorochrome Simulation Program microscopy
.bio-rad.com/.../ fluorophoreDatab.htm
Fluorescence Luminescence Standards www.cstl.ni
st.gov/nist839/ 839.04/luminescence.htm
Analyte Sensor biomed.tamu.edu/obsl/
analyte_fluor.htm
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Fluorescence Polarization www.glycoforum.gr.jp/..
./ GT-C06E.html
Fluorescence Calibration www.pti-nj.com/
tech_8.html
Validation of Comparative Fluorescence www.npl.co
.uk/biotech/ validfluo.html
Extended Depth of Field Imaging www.colorado.edu/
isl/intimages/ fluorescence1.html
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Proteins Heme proteins (hemoglobin, myoglobin,
cytochrome C, hemocyanin) Redox photoproteins
(phytochromes, phycoerythrin, phycocyanin, green
fluorescent protein (http//www.ucmb.ulb.ac.be/...
/ GFP/Fluorophore.gif http//www-bioc.rice.edu/..
./Phillips/ Papers/gfpbio.html
dwb.unl.edu/.../projects/ gmocz/gfp.htm),
phycobiliproteins, rhodopsin, ferredoxins)
Pigments flavins, stilbenes, tryptophan,
tyrosine, purines.. Minerals, metallic chelates
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Metallic chelates Aromatics polyaromatics
(extended ? bonded structures) Crystalline
arrays (intersubband semiconductors)
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Fluorescence Theory www.pharm.uky.edu/ASRG/
HPLC/fluorescence.html
The origins of fluorescence in minerals http//ww
w.uhaul.com/supergraphics/rocks/fluorescence.html
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Fluorescence indicates effective absorbance
re-emission of electromagnetic radiation (often
in the visible range, 300 - 800 nm, but may
involve other ranges, e.g., x - ray. If the
endpoint is presence of fluorescence, e.g.,
qualitative staining of a microscopic specimen,
it can be directly observed or recorded on film
or by digital means. If a quantitative test is
done, color is usually measured with a
spectrofluorimeter.
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Fluorescence Spectrometer Simulation www.inform.u
md.edu/.../ WWW/Fluorescence.html
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Scanning Spectrofluorimeter www.laboratorytalk.co
m/.../ rs_5/rs_5_56.html
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Note the use of light pipes, E,J the multiwell
plate, G the flashlamp, C the photomultiplier
tube, L.
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The Beer Lambert law applies to the initial
absorption of excitation energy but many other
considerations then enter that govern the
efficiency nature of the re-emitted light
observed. Quality factors are associated with
internal energy conversion transfer to nearby
molecules. Quantum efficiency governs the
transition to the meta-stable, pre-fluorescent
excited state this is a measure of the number of
photons emitted relative to the number of photons
exciting the fluorescing structure. Quench
decreases efficiency. Fluorescence, or
fluorescent yield is specific for wavelength,
compound, solvent, pH, temperature.
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Fluorescence scales are arbitrary always
relative to a standard used to calibrate the
instrument. With no background, fluorescence may
reach sensitivity of single molecules.
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From 25-60ºC in 5ºC increments
http//www.isainc.com/fluor/apandtc/tempcont.htm
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www.photobiology.com/photoiupac2000/ karapire/
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Light scattering, F is not directly proportional
to c Photochemical reactions, the system is
dynamic Quenching, multiple compounds absorb
light energy alter spectral intensities acting
on other compounds present http//members.tripod.c
om/obsil/fluor.html Nonlinearity or spectral
impurity of light sources, illumination intensity
varies by wavelength
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Other examples www.firstrays.com/ spectra.htm
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Infrared Optical Masers www.coseti.org/
schawlow.htm
www.phy.davidson.edu/.../ diode_laser.htm
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Microscopy Primer micro.magnet.fsu.edu/.../
techniques/contrast.html
Confocal Microscopy Primers www.cas.muohio.edu/..
./microscopes/ confocal.html web.ncifcrf.gov/rtp/
ial/cml/details.asp http//www.emsl.pnl.gov/homes/
csd/sms/subhomes/r4.html
Confocal Microscopy Confocal Simulator
www.biosci.ohio-state.edu/.../ sackmicroscopy.htm
Imaging Techniques using Fluorescence www.bris.ac
.uk/synaptic/info/ imaging/imaging_1.ht www.imagin
gresearch.com/.../ Fluorescence.asp
Total Internal Reflectance Microscopy http//www.
microscopyu.com/articles/fluorescence/tirf/tirfint
ro.html
Chromosome Spreads with Different Stains
amba.charite.de/cgh/ img/01/img01.html
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Multi-photon Excitation Microscopy
Http//www.loci.wisc.edu/images/ jpegs/amos.jpeg
Lens Focus Formulae http//www.biodevice.com/op
tical_and_fluorescence_formula.htm
Filter Selection Demo http//www.omegafilters.com
/front/curvomatic/spectra.php
Dye Selection http//www.promega.com/geneticidpro
c/ussymp8proc/21.html
FISH http//www.devicelink.com/ivdt/archive/97/01
/005.html
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Fluorescence Image Gallery http//www.nobel.se/ph
ysics/educational/microscopes/fluorescence/gallery
/8.html
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Time Resolved Fluorescence (TRF) www.utu.fi/resea
rch/residues/ methods.html
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Fluorescence Activated Cell Sorting
(FACS) www.bioteach.ubc.ca/.../ FlowCytometry/
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Fluorescence Resonance Energy Transfer
(FRET) www.neuro.mpg.de/research/ scn/celldyn/
laxmi.nuc.ucla.edu8248/.../ part1/en_sens6.html
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Link catalogs of fluorescent microscopy images
turing.bear.uncw.edu/.../ vertebrate.asp
www2.uerj.br/micron/atlas/ atlasenglish/lymph/
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Luminescence www.deakin.edu.au/.../
luminescence.htm pcml.univ-lyon1.fr/Luminophores/
Luminophores.html www.dcb.unibe.ch/.../guedel/
research/hug_lum.htm www.chem.lsu.edu/.../spec/
spectroscopy.html ice.chem.wisc.edu/.../light/
lightandcolor6.html
Luminescence Theory www.shu.ac.uk/.../tutorials/
molspec/lumin1.htm www.tf.uni-kiel.de/.../kap_5/
backbone/r5_1_2.html www.chemistry.nmsu.edu/.../
Lab6/intro.html
Chemiluminescence www.chm.bris.ac.uk/.../
fleming/intro.htm www.lumigen.com/
www.turnerbiosystems.com/.../ 998_2620.html www.s
cienceofspectroscopy.info/ theory/ADVANCE...
www.chm.bris.ac.uk/.../ fleming/mechanism.htm
Lanthanides www.cchem.berkeley.edu/
knrgrp/lantha.html www.chem.ox.ac.uk/icl/heyes/
LanthAct/L9.html web.chem.ufl.edu/kschanze/
NIRLED.htm lanthanides.tripod.com/ ln-cl.html
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Testing Quantum Dots www.phy.hw.ac.uk/resrev/
EQUIS/WP5.html www.ifm.liu.se/.../research/
nano/InAsQD.html
Luciferase Assays www.shpromega.com.cn/
65-6921_09.html
Luminescent Organics www.ndt.net/.../wcndt00/pape
rs/ idn105/idn105.htm
Acridinium Esters www.turnerbiosystems.com/
doc/appnotes/s_0017.html www.metachem.co.uk/
Acridium_NHS_Ester.htm
Imidazopyrazinones www.ch.ic.ac.uk/.../echet96/
papers/055/ref/6.html
Dioxetanes www.michdiag.com/ app.html
Photoluminescence www.fz-rossendorf.de/FWI/
FWIM/lumi_photo_e.htm
Bioluminescence bioluminescence.free.fr/
dosatp.htm
Cathodoluminescence plaza.snu.ac.kr/lee2602/
atlas/cath_intro.html
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Scanning Tunneling Luminescence level2.phys.strat
h.ac.uk/ ssd/HTML/charhtml/stl.ht
Oxygen Electrodes www.nogi.ch/chemistry/
oxygensensor.html chemistry.mtu.edu/.../thesis/
ch02/Chapter02.html
QuickTime Chemistry Movies www.shsu.edu/chm_tgc/
sounds/sound.html
Chemiluminescence Detection in HPLC www.ftns.wau.
nl/.../lunteren/ lunteren.htm
Multimode Plate Reader www.image1.com/resources/
insights/0306/analyst.cf
Materials Characterization www.ifm.liu.se/.../pop
ular/ Chapter4.html
Biological Toxic Metal Assays www.aboatox.com/
environmental_analysis.html
Biospore Testing www.nasatech.com/Briefs/
Mar03/NPO30469.html
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Use polycyclic arene esters to produce
luminescence in almost any visible color. H. Bock
co-authors report that they used the good
charge-transport properties of columnar liquid
crystals of these compounds to produce
light-emitting diodes (LEDs) with color-tuned
electro-luminescence. E.g., they used
perylene-3,4,9,10-tetracarboxylic acid ester 1 to
form an LED with orange red luminescence, while
a series of esters of the triphenylene derivative
2 produced luminescence in the violetblue
range. R is methyl through n-octyl. By varying
the arene core the R group on the esters, the
authors demonstrated photoluminescence
wavelengths over the entire visible spectrum.
Particularly interesting was their use of
multiple layers of different esters to form LEDs
with almost white luminescence. (Angew. Chem.,
Int. Ed. 2001, 40, 20602063 WJP)
pubs.acs.org/.../ci/31/ i12/html/12hc3.html
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www.biodevice.com/ FluorescenceChemiluminescen...
Luminol Reaction www.newlisbon.k12.wi.us/
reactions/home.html234
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Bioluminescent Organisms www.mcb.harvard.edu/.../
bioluminescence.html
Bioluminescent Chemistries http//www.turnerbios
ystems.com/ doc/appnotes/pro_bio.htm
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Temperature Effects on Chemiluminescence dbweb.li
v.ac.uk/ltsnpsc/ practs/djm3.htm
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Bioaffinity Flow Analyzer www.analykem.lu.se/home
page/ res/res3.htm
Suppliers www.vectorlabs.com/ products.asp?catID
157loc...