Beruflich Dokumente
Kultur Dokumente
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 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
Naturally Fluorescent Molecules
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..
Metallic chelates
Aromatics & polyaromatics (extended
bonded structures)
Crystalline arrays (intersubband
semiconductors)
Theory of Fluorescence
Fluorescence Theory:
www.pharm.uky.edu/ASRG/ HPLC/fluorescence.html
http://www.ucd.ie/app-phys/ http://www.shu.ac.uk
michael/ photosyn.html /.../tutorials/
Scanning Spectrofluorimeter:
www.laboratorytalk.com/.../ rs_5/rs_5_56.html
Microplate Spectrofluorometer
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.
Spectrofluorimetry: Basics
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.
Spectrofluorimetry
Fluorescence scales are arbitrary & always
relative to a standard used to calibrate the
instrument. With no background, fluorescence
may reach sensitivity of single molecules.
Impacts of pH on
Fluorescence
Impacts of Temperature on Fluorescence
www.photobiology.com/photoi
upac2000/
karapire/
Complications for Spectrofluorimetry
Light Source
Intensity
Variation by
Wavelength
www.pti-nj.com/ obb_spectra.html
www.phy.davidson.edu/.../ diode_laser.htm
Other Applications of Fluorescence
Fluorescence Microscopy
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/fluores
cence/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
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/micros
copes/fluorescence/gallery/8.html
FISH, Human Cells
Dividing CHO Cell
Zeiss
P. Bjrk, 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
http://www.chemie.uni-
marburg.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/
Types of Luminescence
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 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 pubs.acs.org
almost white luminescence. (Angew. Chem.,
/.../ci/31/ i12/html/12
Int. Ed. 2001, 40, 20602063; WJP)
hc3.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
Bioluminescent
Chemistries:
http://www.turne
rbiosystems.co
m/
doc/appnotes/pr
o_bio.htm
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...