Fluorescence Spectroscopy, Imaging and Probes : New Tools in Chemical, Physical, and Life Sciences

by Conference on Methods and Applications of Fluorescence: Spectroscopy, Imaging, and Probes (7th : 2001 : Amsterdam, Netherlands); Visser, Antonie J. W. G.; Gerritsen, Hans C.; Kraayenhof, Ruud; Kraayenhof, Ruud; Visser, Antonie J. W. G.

• ISBN13: 9783540427681
• ISBN10: 3540427686
• Format: Hardcover
• Copyright: 2002-11-01
• Publisher: Springer Verlag

Summary

The application of fluorescence in drug discovery, high-throughout screenings in genomics and proteomics is and will be evidently successful. The increased use of fluorescence techniques is greatly enhanced by the improved instrumentation pioneered by inventive scientists and now made available commercially by several high-tech companies. Moreover, the design and development of many new molecular probes with higher selectivity for specific microenvironmental properties has stimulated many new researchers to employ fluorescence techniques for solving their problems. Probably the most significant breakthrough in fluorescence is its use in detection of single molecules and even of their real-time dynamics. Also, probing inside living cells has become a hot topic in the life sciences. This topic book reflects the updates of scientific progress presented by frontline researchers.

Table of Contents

Part 1: Fluorescence Spectroscopy: New Approaches and Probes
Advanced Luminescent Labels, Probes and Beads and their Application to Luminescence Bioassay and Imaging	3	(40)
O. S. Wolfbeis
M. Bohmer
A. Durkop
J. Enderlein
M. Gruber
I. Klimant
C. Krause
J. Kurner
G. Liebsch
Z. Lin
B. Oswald
M. Wu
Introduction	4	(2)
A General Logic for Designing Fluorescent Cyanine Type Probes and Labels of Defined Color	6	(2)
Diode Laser-excitable Probes for General Protein Detection	8	(2)
Diode Laser-compatible Amino-Reactive Covalent Labels	10	(1)
Diode Laser-assisted Fluorescent Single Molecule Detection of Dyes and Labeled Proteins	11	(3)
New Labels for Flow Cytometric Determination of HSA	14	(1)
Diode Laser-excitable DNA Labels	15	(1)
New Resonance Energy Transfer Gene Assays	16	(1)
Reactive Ruthenium Ligand Complexes as Markers for Bioassays	17	(1)
Diode Laser-excitable Fluorescent Polymer Beads	18	(2)
Polyaniline-Coated Nano-beads (~200 nm in Diameter) with pH-dependent Fluorescence	20	(2)
Phosphorescent Poly(Acrylonitrile) Nanospheres (10-100 nm in Ø) as Markers for Optical Assays	22	(3)
Competitive Binding of Streptavidin to Biotinylated Nanobeads as Studied by Resonance Energy Transfer	25	(2)
Nanobeads as Reference Dyes in Luminescent Lifetime Imaging Using DLR	27	(3)
Phosphorescent Nanospheres for Use in Advanced Time-resolved Multiplexed Bioassays (λ, τ)	30	(4)
Beads Dyed with a Europium-based Label and Excitable with the 405-nm LED Diode Laser	34	(1)
A Europium (III)-based Probe for Use in Oxidase-Associated Reactions	35	(8)
Significance of Probes for Hydrogen Peroxide (HP)	35	(1)
A New Probe for Hydrogen Peroxide	35	(1)
Glucose Assay Using the Europium Probe	36	(1)
Peroxidase Assay Using the Europium Probe	37	(1)
Catalase	38	(1)
A New Enzyme-linked Immunosorbent Assay (ELISA)	38	(2)
References	40	(3)
Fluorescence Spectral Engineering-Biophysical and Biomedical Applications	43	(26)
J. R. Lakowicz
I. Gryczynski
Y. Shen
J. Malicka
S. D'Auria
Z. Gryczynski
Introduction	44	(12)
Fluorescence Spectral Engineering	45	(2)
Overview of Metallic Surface Effects on Fluorescence	47	(3)
Theory for Fluorophore Metal Interactions	50	(4)
Spatial Distribution of Emission Near Metal Surfaces	54	(1)
Resonance Energy Transfer	55	(1)
Experimental Results on Fluorophore--metal Interactions	56	(9)
Silver Island Films and Experimental Geometry	56	(1)
Effects of Silver Island Films on Emission Spectra of Rhodamine B and Rose Bengal	56	(2)
Effect of Silver Island Films on Photostability	58	(1)
Effects of Silver Island Films on the Lifetime of Rhodamine B and Rose Bengal	59	(1)
Effects of Quantum Yield on Silver Island Enhancements	60	(1)
Effect of Silver Island on Intrinsic Protein Fluorescence	61	(1)
Effects of Silver Island on Nucleic Acid Bases and DNA	62	(1)
Effects of Silver Island on Resonance Energy Transfer	63	(2)
Conclusion	65	(4)
References	65	(4)
Fluorescence Nanometrology in Sol-Gels	69	(18)
D.J.S. Birch
C.D. Geddes
J. Karolin
R. Leishman
O.J. Rolinski
Introduction	70	(2)
Sol-gel Chemistry	72	(3)
Anisotropy Theory	75	(1)
FRET Donor-acceptor Distribution Theory	76	(2)
Acidic Hydrogels	78	(1)
Alkaline Hydrogels	79	(1)
Alkoxide Alcogels	80	(1)
Wet Pore Metrology	81	(2)
Conclusions	83	(4)
References	84	(3)
Integrated Supramolecular Systems: From Sensors to Switches	87	(14)
J.-P. Malval
I. Gosse
J.-P. Morand
R. Lapouyade
Introduction	88	(2)
Optical Detection of Ion-Ground State Probe Interaction	90	(1)
Cation Sensing From Fluorescent Photoinduced Intramolecular Charge Transfer (PICT) Sensors	91	(5)
Optical and Electrochemical Release of Cations	96	(1)
Conclusions	97	(4)
References	98	(3)
Ratiometric Probes: Design and Applications	101	(10)
A. P. Demchenko
A. S. Klymchenko
V. G. Pivovarenko
S. Ercelen
Introduction	102	(1)
3-Hydroxyflavones and Other 3-Hydroxychromone Derivatives. New Compounds and Their Properties	103	(1)
A New Level of Sensitivity to Solvent Polarity	104	(2)
Amplification by ESIPT of Electrochromic Effects	106	(1)
Molecular Order and Dynamics in Phospholipid Membranes	107	(1)
Amplification by ESIPT of Site-selective Red Edge Effect	108	(1)
Conclusions	109	(2)
References	110	(1)
Binding of Ethidium to Yeast tRNA phe: A New Perspective on an Old Bromide	111	(12)
M. Tramier
O. Holub
J. C. Croney
T. Ishi
S. E. Seifried
D. M. Jameson
Overview	112	(1)
Experimental	112	(4)
Sample Preparation	112	(1)
Multifrequency Phase and Modulation Fluorometry	112	(1)
Three Component Analysis	113	(3)
New Analysis	116	(2)
The Model	118	(1)
Effect of lonic Strength	118	(1)
Conclusions	119	(4)
References	120	(3)
Experimental Aspects of DNA Computing by Blocking: Use of Fluorescence Techniques for Detection	123	(8)
K. A. Schmidt
C. V. Henkel
G. Rozenberg
H. P. Spaink
Introduction	124	(1)
Experimental	124	(1)
Results and Discussion	125	(2)
Conclusion	127	(4)
References	128	(3)
Part 2 Fluorescence Spectroscopy of Single Molecules and Molecular Assemblies
Multiparametric Detection of Fluorescence Emitted from Individual Multichromophoric Systems	131	(22)
M. Cotlet
J. Hofkens
M. Maus
F. C. De Schryver
Introduction	132	(1)
Materials and Methods	133	(3)
Sample Preparation	133	(1)
Experimental Set-up	133	(3)
Data Processing and Analysis	136	(5)
Pulsed Excitation	136	(3)
CW Excitation	139	(2)
Results and Discussion	141	(8)
Pulsed Excitation	142	(3)
CW Excitation	145	(4)
Conclusions	149	(4)
References	150	(3)
Fluorescence Intensity Distribution Analysis (FIDA) and related fluorescence fluctuation techniques: theory and practice	153	(30)
P. Kask
C. Eggeling
K. Palo
U. Mets
M. Cole
K. Gall
Introduction	154	(2)
Instrumentation for Fluorescence Fluctuation Spectroscopy	156	(2)
Assumptions and Conventions	158	(2)
Generating Functions for FIDA and Related Methods	160	(2)
FIDA	162	(3)
2D-FIDA	165	(3)
FIMDA	168	(4)
FILDA	172	(6)
Conclusions	178	(5)
References	178	(5)
Single Molecule Reactions of the Enzyme LDH and of Restriction Endonucleases in the Fluorescence Microscope	183	(14)
B. Nasanshargal
B. Schafer
K. O. Greulich
Introduction	184	(1)
Femtodroplets and the Poisson Statistics	185	(1)
From Concentrations to Intermolecular Distances	186	(1)
Why Fluorescence Microscopy?	187	(1)
Single Molecule Enzyme Reactions with Small Substrates	187	(4)
Restriction Endonuclease Reactions	191	(3)
Conclusions	194	(3)
References	195	(2)
Monitoring γ-Subunit Movement in Reconstituted Single EF0F1 ATP Synthase by Fluorescence Resonance Energy Transfer	197	(14)
M. Borsch
M. Diez
B. Zimmermann
R. Reuter
P. Graber
Introduction	198	(1)
Visualizing Intersubunit Rotation	198	(2)
FRET-labeled F0F1 ATP Synthase EF0-b64-TMR-F1-γ106-Cy5	200	(6)
Synthezising ATP with Reconstituted EF0-b64-TMR-F1-γ106-Cy5	201	(1)
Set-up for Single Enzyme FRET Analysis	201	(2)
Discrimination of Three γ-Subunit Positions with AMPPNP	203	(2)
Unidirectional Motion of the γ-Subunit with ATP	205	(1)
Conclusions	206	(5)
References	207	(4)
Part 3 Application of Fluorescence in Biological Membrane and Enzyme Studies
Application of the Wavelength-selective Fluorescence Approach to Monitor Membrane Organization and Dynamics	211	(14)
A. Chattopadhyay
Introduction	212	(1)
Red Edge Excitation Shift (REES)	212	(2)
The Wavelength-selective Fluorescence Approach	214	(1)
The Wavelength-selective Fluorescence Approach: A Novel Tool to Monitor Organization and Dynamics of the Membrane Interfacial Region	214	(3)
Wavelength-selective Fluorescence as a Membrance Dipstick	217	(1)
Application of the Wavelength-selective Fluorescence Approach to Membrane Peptides and Proteins	218	(2)
Wavelength-selective Fluorescence in Micelles	220	(1)
Conclusions	221	(4)
References	221	(4)
Fluorescence Approaches for the Characterization of the Peripheral Membrane Binding of Proteins Applied for the Blood Coagulation Protein Prothrombin	225	(16)
R. Hutterer
M. Hof
Introduction	226	(1)
Prothrombin Binding to Negatively Charged Membrane Surface Characterized by Protein Fluorescence	227	(6)
Intrinsic Protein Fluorescence: Picosecond Tryptophan Fluorescence of Membrane-bound Prothrombin Fragment 1 (F1)	227	(4)
Overview on Investigations Applying Fluorescently Labeled Prothrombin	231	(2)
Prothrombin-induced Changes in the Organization of Phospholipid Bilayers	233	(5)
Solvent Relaxation (SR)	233	(1)
Solvent Relaxation Probed by the Headgroup Labels Prodan and Patman	233	(2)
Influence of Prothrombin and its Fragment 1 on the Phospolipid Headgroup Organization	235	(2)
Fluorescence Anisotropy: Influence of Prothrombin and its Fragment 1 on PC/PS Membrane Order	237	(1)
Pyrene Fluorescence: Influence of Fragment 1 on Membrane Order Monitored by the Excimer/Monomer Ratio	237	(1)
Conclusions	238	(3)
References	239	(2)
Assessment of Membrane Fluidity in Individual Yeast Cells by Laurdan Generalised Polarisation and Multi-photon Scanning Fluorescence Microscopy	241	(12)
R. P. Learmonth
E. Gratton
Introduction	242	(2)
Yeast Membrane Fluidity	242	(1)
Multi-photon Scanning Fluorescence Microscopy	243	(1)
Determination of Membrane Fluidity Using Laurdan Generalized Polarization	244	(1)
Materials and Methods	244	(2)
Results and Discussion	246	(4)
Conclusions	250	(3)
References	251	(2)
Formation of Higher Order Signal Transduction Complexes as Seen by Fluorescence Spectroscopy	253	(10)
L. Dowal
S. Scarlata
Introduction	254	(2)
Experimental	256	(7)
Membrane Binding	257	(1)
Interactions of PLCβ and G Protein Subunits on Membrane Surface	257	(1)
PLCβ2-Gβy Associations on Membrane Surfaces	258	(1)
Effects of Lipid Rafts on Protein Association	259	(1)
References	260	(3)
Mechanisms of the Modulation of Membrane Interfacial Enzyme Catalysis by Non-lamellar Forming Lipids: Comparison with the Behavior of a Fluorescent Probe in Membranes	263	(14)
R.M. Epand
R. Cornell
S. M.A. Davies
R. Kraayenhof
Introduction	264	(2)
Specific vs. Non-specific Modulation of Protein Activity	264	(1)
Non-lamellar Forming Lipids	265	(1)
Specific Protein Examples	266	(2)
Protein Kinase C	266	(1)
Phosphocholine Cytidylyltransferase	267	(1)
Molecular Mechanisms of Enzyme Activation by Non-lamellar Forming Lipids	268	(1)
Curvature Strain	268	(1)
Lateral Pressure Profile	269	(1)
Other Mechanisms	269	(1)
Criteria for the Role of Membrane Curvature Strain	269	(2)
Cubic Phases	269	(2)
Homologous Lipids	271	(1)
Relationship of the Properties of a Fluorescent Interfacial Membrane Probe	271	(1)
Comparison of the Mechanisms of Activation of PKC and CT by Non-lamellar Forming Lipids	272	(1)
Conclusions	273	(4)
References	274	(3)
Emission Spectroscopy of Complex Formation between Escherichia coli Purine Nucleoside Phosphorylase (PNP) and Identified Tautomeric Species of Formycin Inhibitors Resolves Ambiguities Found in Crystallographic Studies	277	(20)
B. Kierdaszuk
Introduction	278	(2)
Formycin A and its N-methyl Analogs, Specific Inhibitors of E. coli PNP	280	(1)
Ambiguities Found in the Crystallographic Structure of Enzyme-Ligand Complex	281	(2)
Solution Structure of Inhibitors Bound by the Enzyme	283	(8)
Tautomeric Equilibria, and Absorption and Emission Spectra of the Tautomeric Species in Solution	283	(3)
Shifts Between Absorption and Emission Spectra of the Enzyme and Ligands	286	(1)
Effect of Binding of the Identified Tautomeric Species on their Fluorescence and Phosphorescence	287	(4)
Conclusions	291	(6)
References	292	(5)
Part 4 Microscopic Imaging Techniques and their Application for the Study of Living Cells
Fluorescence Lifetime Imaging Implemented with Resonant Galvanometer Scanners	297	(20)
J. J. Birmingham
Introduction	298	(1)
Theory	299	(6)
Simulations	305	(3)
Experimental Set-up	308	(3)
Results	311	(3)
Conclusions	314	(3)
References	315	(2)
Spectral Imaging of Single CdSe/ZnS Quantum Dots Employing Spectrally- and Time-resolved Confocal Microscopy	317	(20)
W.G.J.H.M. Van Sark
P.L.T.M. Frederix
M.A.H. Asselbergs
D.J. Van Den Heuvel
A. Meijerink
H.C. Gerritsen
Introduction	318	(1)
Experimental	319	(6)
Spectrography-CLSM Set-up	319	(1)
Description	319	(3)
Performance	322	(2)
QD Synthesis and Characterization	324	(1)
Results and Discussion	325	(7)
Conclusion	332	(5)
References	332	(5)
Imaging of Oxidative Stress in Plant Cells by Quantitative Fluorescence Microscopy and Spectroscopy	337	(12)
J. W. Borst
M. A. Uskova
N. V. Visser
A. J. W. G. Visser
Introduction	338	(2)
Experimental Procedures	340	(2)
Tobacco BY-2 Cells	340	(1)
Preparation of Cell Suspension	340	(1)
Protoplast Isolation	341	(1)
Labeling Cells with BP-C11	341	(1)
Confocal Microscopy	341	(1)
Steady-state Fluorescence	342	(1)
Results and Discussion	342	(4)
Measurements and Imaging of Oxidative Stress in Tobacco BY-2 Cells	342	(2)
Effect of Inhibitors and ROS Scavengers on the Hydrogen Peroxide Production in BY-2 Cells	344	(2)
Conclusions	346	(3)
References	347	(2)
The Biomedical Use of Rescaling Procedures in Optical Biopsy and Optical Molecular Imaging	349	(12)
O. Minet
J. Beuthan
K. Licha
C. Mahnke
Introduction	350	(1)
Method of Rescaling	350	(2)
Biomedical Examples	352	(6)
Optical Biopsy in the UV Range Using Endogenous Chromophores	352	(1)
Experiment	353	(1)
Results	354	(1)
Optical Molecular Imaging in the NIR Range Using Exogenous Contrast Agents	354	(2)
Experiment	356	(1)
Results	357	(1)
Discussion and Conclusions	358	(3)
References	359	(2)
Looking into a Living Cell	361	(12)
M. Van Borren
N. R. Brady
J. Ravelsloot
H. V. Westerhoff
Introduction	362	(1)
How to Choose your Fluorescent Indicator	362	(2)
Process of Interest	362	(1)
Ratiometric Dyes	363	(1)
Buffering Power	364	(1)
Photo-toxicity	364	(1)
Considerations Concerning the Experimental Approach	364	(2)
Selective Loading	364	(2)
Additional Loading Techniques	366	(1)
The Imaging System	366	(1)
Examples of Fluorescence Microscopy in Living Cells	366	(5)
Detection of the Ca2+ Ion	366	(1)
Determination of Intracellular pH	367	(1)
Mitochondrial Energetics	368	(2)
Reactive Oxygen Species (ROS)	370	(1)
Autofluorescence: Marker of Redox State	371	(1)
Conclusions	371	(2)
References	371	(2)
Expression of Multicolor Fluorescent Fusion Proteins in Zebrafish Cell Cultures: A Versatile Tool in Cell Biology	373	(8)
C. K. D. Breek
F. Van Iren
S. E. Wijting
N. Stuurman
H. P. Spaink
Introduction	374	(1)
Zebrafish Cell Lines	374	(2)
Microscopical Analysis	376	(1)
Analysis of Transfected Cell Lines	377	(2)
Conclusions	379	(2)
References	379	(2)
Subject Index	381