Fluorescence Spectroscopy in Biology: Advanced Methods and their Applications to Membranes, Proteins, DNA, and Cells (Springer Series on Fluorescence, 3)

Volume 3 of this new series focuses on brandnew research and applications in biology, biophysics and other fields of life sciences. Many frontline researcher have contributed to this highly attractive and interdisciplinary volume which spans the entire field of present fluorescence spectroscopy including nanotechnology, membrane and DNA studies and fluorescence imaging in cancer research. Contents......Page 8 Part 1 Fluorescence Spectroscopy: Basics and Advanced Approaches......Page 19 1.1 Introduction......Page 20 1.2 Fluorescence and its Measurement......Page 21 1.3 Polarized Fluorescence......Page 24 1.4 Influence of Fluorescence Quenching......Page 28 1.5 Influence of Solvent Relaxation on Solute Fluorescence......Page 34 1.6 Fluorescence Resonance Energy Transfer as a Spectroscopic Ruler......Page 37 1.7 Irreversible Photobleaching......Page 39 1.8 Single Molecule Fluorescence......Page 40 1.9 Optical Sensors Based on Fluorescence......Page 41 References......Page 42 2.1 Introduction......Page 47 2.2 General Principles of Time-Resolved Fluorometry......Page 48 2.3 Pulse Fluorometers......Page 52 2.4 Phase-Modulation Fluorometers......Page 54 2.5 Data Analysis......Page 58 2.6 Specific Applications......Page 59 2.7 Concluding Remarks......Page 64 References......Page 65 3.1 Introduction......Page 66 3.2 Theory......Page 67 3.4 Results and Discussion......Page 68 References......Page 71 4.1 Introduction......Page 73 4.2 Fluorescence Resonance Energy Transfer......Page 74 4.3 FRET Sensors......Page 75 4.4 Fluorescence Nanotomography Theory......Page 77 4.5 Experimental......Page 83 References......Page 86 5.2 Basic Principles of the SR Method......Page 88 5.3 Applications of the SR Technique by Using Time-Correlated Single Photon Counting......Page 90 References......Page 94 6.1 Introduction......Page 96 6.2 Combination of TIRFM with Other Methods......Page 100 6.3 Advanced Topics......Page 108 6.4 Other Applications......Page 112 6.5 Summary......Page 115 References......Page 116 7.1 Introduction......Page 121 7.2 Photophysics, Probes and Markers......Page 122 7.3 Physical Techniques......Page 126 7.4 Data Acquisition and Evaluation......Page 133 References......Page 139 Part 2 Application of Fluorescence Spectroscopy to Biological Membranes......Page 148 8.1 Introduction......Page 149 8.2 Identification of Lipid Compositions Forming Rafts......Page 150 8.3 Temperature Dependence in Domain/Raft Formation......Page 155 8.4 Affinity of Lipids and Proteins for Rafts as Detected by Quenching......Page 159 8.5 Alternative Fluorescence Methods for the Detection of Rafts......Page 163 References......Page 165 9.1 Introduction......Page 166 9.3 Domains in Membranes......Page 167 9.4 Fluorescent Probes: Advantages and Disadvantages......Page 169 9.5 Correlation with Other Experimental Techniques......Page 171 9.6 Concluding Remarks and Future Directions......Page 173 References......Page 174 Part 3 Application of Fluorescence Spectroscopy to Protein Studies......Page 176 10.1 Introduction......Page 177 10.2 Dynamic Fluorescence of Tryptophan......Page 178 10.3 Time-Resolved Fluorescence Resonance Energy Transfer (tr-FRET) in Protein Folding......Page 187 References......Page 191 11.1 Introduction......Page 194 11.2 Materials and Methods......Page 197 11.3 Results and Discussion......Page 199 11.4 Conclusion......Page 207 References......Page 209 Part 4 Application of Fluorescence Spectroscopy to DNA and Drug Delivery......Page 212 12.1 Introduction to Non-Viral Gene Therapy and its Development......Page 213 12.2 Using Fluorescence Techniques to Determine the Efficiency of DNA Condensing Agents: an Important First Step in the Mechanism of NVGT......Page 216 12.3 Conjugation of Lipopolyamines to Fluorophores: Probes Derived from DNA Delivery Agents......Page 218 12.4 Preparation of Fluorescent Macromolecules......Page 221 12.5 Lipopolyamines and Cationic Lipids Used in Transfection......Page 223 12.6 Association and Dissociation Studies of DNA Complexes Through Fluorescence Correlation Spectroscopy (FCS)......Page 227 12.7 DNA Complexes and Their Intracellular Trafficking: Monitoring by Fluorescence (Förster) Resonance Energy Transfer (FRET)......Page 228 12.8 Fluorescence Microscopy in NVGT......Page 229 12.9 Conclusions......Page 235 References......Page 236 13.1 Introduction......Page 241 13.2 Fluorescence Techniques as Tools for the Development of Targeted Drug Delivery Systems......Page 243 13.3 Perspectives......Page 251 References......Page 252 Part 5 Fluorescence Spectroscopy in Cells: FCS and Quantum Dots......Page 255 14.1 Introduction......Page 256 14.2 Fluorescence Correlation Spectroscopy Step by Step......Page 258 14.3 Cellular FCS......Page 266 14.4 Perspectives......Page 271 References......Page 272 15.1 Introduction......Page 274 15.2 Photophysical properties of Quantum Dots......Page 275 15.3 Applications of Quantum Dots as Fluorescent Probes......Page 280 References......Page 284 16.1 Introduction......Page 286 16.2 Experiments......Page 288 16.3 Results......Page 291 16.4 Discussion......Page 296 References......Page 297 A......Page 299 C......Page 300 D......Page 302 E......Page 303 F......Page 304 G......Page 305 I......Page 306 L......Page 307 M......Page 308 N......Page 309 P......Page 310 Q......Page 311 S......Page 312 V......Page 314 Z......Page 315 Take any combination of the following features: supramolecular structures with a specific fluorescent probe localized as you would like; nanoscale spatial reso- tion; tailor-made molecular and/or solid-state fluorescing nanostructures; us- friendly and/or high- throughput fluorescence techniques; the ability to do wh- ever you wish with just one single (supra)molecule; utilization of non-linear optical processes; and, last but not least, physical understanding of the processes resu- ing in a (biological) functionality at the single molecule level. What you will then have is some recent progress in physics, chemistry, and the life sciences leading to the development of a new tool for research and application. This was amply demonstrated at the 8th Conference on Methods and Applications of Fluorescence: Probes, Imaging, and Spectroscopy held in Prague, the Czech Republic on August 24th-28th, 2003. This formed a crossroad of ideas from a variety of natural science and technical research fields and biomedical applications in particular. This volume - the third book in the Springer-Verlag Series on Fluorescence - reviews some of the most characteristic topics of the multidisciplinary area of fluorescence applications in life sciences either presendted directly at th 8th MAF Conference or considered to be a cruical development in the field. In the initial contribution in Part 1 - Basics and Advanced Approaches, the - itors explain the basics of fluorescence and illustrate the relationship between some modern fluorescence techniques and classical approaches. The second contrigution by B. Basics of fluorescence spectroscopy : basics and advanced approaches Pulse and phase fluorometries : an objective comparison Non-exponential fluorescence of electronically coupled donors contains distance information Fluorescence nanotomography : recent progress, constraints and opportunities Solvent relaxation as a tool for probing micro-polarity and fluidity Total internal reflection fluorescence microscopy : applications to biophysics Single molecule spectroscopy : basics and applications Raft microdomains in model membranes as revealed by fluorescence quenching The lateral structure of lipid membranes as seen by fluorescence microscopy Protein dynamics and protein folding dynamics revealed by time-resolved fluorescence Time-resolved fluorescence and two-photon FCS investigation of the interaction of HIV-1 nucleocapsid protein with hairpin loop oligonucleotides Fluorescence techniques in non-viral gene therapy Fluorescence applications in targeted drug delivery Fluorescence correlation spectroscopy in cell biology Fluorescence quantum dots : properties and applications Heat stress of cancer cells : fluorescence imaging of structural changes with quantum dots 605 and Alexa 488. After the pioneering works of Kasha, Vavilov, Perrin, Jabloski, Weber, Stokes and Förster (including the appearance of the first book of the latter on fluorescence of organic molecules in 1951 [1]), fluorescence spectroscopy became a widely used scientific tool in biochemistry, biophysics and material sciences. "This volume spans the entire field of present flourescence spectroscopy including nanotechology, membrane and DNA studies and fluorescence imaging in cancer research."--Résumé de l'éditeur