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.”
Stedman’s Medical Dictionary, 24 th ed., 1982.

http://chsfpc5.chem.ncsu.ed
u/Poznan/membrane_fuse/in dex.htm
micro.magnet.fsu.edu/.../ jablonski/lightandcolor/

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.nist.gov/nist839/ 839.04/luminescence.htm
Analyte Sensor: biomed.tamu.edu/obsl/ analyte_fluor.htm

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

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://wwwbioc.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



Fluorescence Theory: www.pharm.uky.edu/ASRG/ HPLC/fluorescence.html
The origins of fluorescence in minerals: http://www.uhaul.com/supergraphics/rocks/fluorescence.
html

Fluorescence Spectrometer Simulation: www.inform.umd.edu/.../ WWW/Fluorescence.html
http://www.ucd.ie/appphys/michael/ photosyn.html
http://www.shu.ac.uk/.../ tutorials/

Scanning Spectrofluorimeter: www.laboratorytalk.com/.../ rs_5/rs_5_56.html

Http://www.moleculardevices.com/ media/gem_xs_optics.gif
Note the use of light pipes, E,J; the multiwell plate, G; the flashlamp, C; & the photomultiplier tube, L.

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.

Fluorescence scales are arbitrary & always relative to a standard used to calibrate the instrument. With no background, fluorescence may reach sensitivity of single molecules.

From 2560ºC in 5ºC increments: http://www.isainc.com/fluor/apandtc/tempcont.htm

www.photobiology.com/photoi upac2000/ karapire/

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.com/obsil/fluor.html
Nonlinearity or spectral impurity of light sources , illumination intensity varies by wavelength

www.physics.montana.edu/
.../ emissionspectra.html
www.pti-nj.com/ obb_spectra.html
Other examples: www.firstrays.com/ spectra.htm

Infrared & Optical Masers: www.coseti.org/ schawlow.htm
Laser Theory: www.physics.ubc.ca/.../p420_95/ mark/htheory.html
www1.union.edu/.../ laser_theory.htm
webphysics.davidson.edu/.../ laserlab/theory.htm
people.deas.harvard.edu/.../ ls2_unit_6.html
www.people.fas.harvard.edu/.../ chemlaser.htm
www.phy.davidson.edu/.../ diode_laser.htm

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.imagingresearch.com/.../ Fluorescence.asp
Total Internal Reflectance Microscopy: http://www.microscopyu.com/articles/fluor escence/tirf/tirfintro.html
Chromosome Spreads with Different Stains: amba.charite.de/cgh/ img/01/img01.html

Multi-photon Excitation Microscopy:
Http://www.loci.wisc.edu/images/ jpegs/amos.jpeg
Lens & Focus Formulae: http://www.biodevice.com/optical_and_fluorescence_ formula.htm
Filter Selection Demo: http://www.omegafilters.com/front/curvomatic/spectra.php
Dye Selection: http://www.promega.com/geneticidproc/ussymp8proc/21.html
FISH: http://www.devicelink.com/ivdt/archive/97/01/005.html

Fluorescence Image Gallery: http://www.nobel.se/physics/educational/micro scopes/fluorescence/gallery/8.html
FISH, Human Cells
Dividing CHO Cell
Zeiss
P. Björk, Stockholm University
C. elegans Brain
H. Hutter
Max Planck Institut, Heidelberg

Time Resolved Fluorescence (TRF): www.utu.fi/research/residues/ methods.html

Fluorescence Activated Cell Sorting (FACS): www.bioteach.ubc.ca/.../ FlowCytometry/

Fluorescence Resonance Energy Transfer (FRET): www.neuro.mpg.de/research/ scn/celldyn/ laxmi.nuc.ucla.edu:8248/.../ part1/en_sens6.html
Real Time PCR &
Molecular Beacons
(NucliSens)

Fluorogenic response of differently colored molecular beacons to the addition of target. A solution of each molecular beacon was placed in a pair of test tubes. The molecular beacons contained (left to right) coumarin (blue), EDANS (blue-green), fluorescein (green), Lucifer yellow, tetramethylrhodamine (orange), and Texas red. All molecular beacons contain DABCYL as a quencher. Complementary singlestranded oligonucleotides were added to the left tube of each pair, and the tubes were illuminated with a broad-wavelength ultraviolet lamp. (Tyagi et al., 1998).
http://www.biolegio.com/beacons.htm

http://www.chemie.unimarburg.de/~butenuth/642/Lumi
Dateien/bilder/physik5.jpg

Link catalogs of fluorescent microscopy images: turing.bear.uncw.edu/.../ vertebrate.asp
www2.uerj.br/~micron/atlas/ atlasenglish/lymph/

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.scienceofspectroscopy.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

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/papers/ 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

Scanning Tunneling Luminescence: level2.phys.strath.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/.../popular/ Chapter4.html
Biological Toxic Metal Assays: www.aboatox.com/ environmental_analysis.html
Biospore Testing: www.nasatech.com/Briefs/ Mar03/NPO30469.html

Use polycyclic arene esters to produce luminescence in almost any visible color.
H. Bock & co-authors report that they used the good chargetransport properties of columnar liquid crystals of these compounds to produce light-emitting diodes (LEDs) with color-tuned electroluminescence. [ 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 violet –blue 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, 2060 –2063; WJP) pubs.acs.org/.../ci/31/ i12/html/12hc3.html

www.biodevice.com/
FluorescenceChemil uminescen...
Luminol Reaction: www.newlisbon.k12.wi.us/ reactions/home.html%234

Bioluminescent Organisms: www.mcb.harvard.edu/.../ bioluminescence.html
Railroad Worm
Bioluminescent
Chemistries: http://www.turne
rbiosystems.co
m/ doc/appnotes/pr o_bio.htm
Bacteria Squid

Temperature Effects on Chemiluminescence: dbweb.liv.ac.uk/ltsnpsc/ practs/djm3.htm

Bioaffinity Flow Analyzer: www.analykem.lu.se/homepage/ res/res3.htm
Suppliers: www.vectorlabs.com/ products.asp?catID=157&loc...