Closing a gap in the literature, this handbook gathers all the information on single particle tracking and single molecule energy transfer. It covers all aspects of this hot and modern topic, from detecting virus entry to membrane diffusion, and from protein folding using spFRET to coupled dye systems, as well recent achievements in the field. Throughout, the first-class editors and top international authors present content of the highest quality, making this a must-have for physical chemists, spectroscopists, molecular physicists and biochemists.
Christoph Brauchle is Professor at the Ludwig-Maximilian-University (LMU) Munich. After his PhD, he spent one year as a postdoc at IBM in San Jose, California, USA. His research focuses on imaging, spectroscopy and manipulation of single molecules in bio- and nano-sciences. Besides numerous publications in international journals, Prof. Brauchle has won several honors, including the prestigious Philip Morris Research award and the Karl Heinz Beckurts Prize 2002. He also is a member of the Academia Europaea. Don C. Lamb is Professor for Biophysical Chemistry at the LMU Munich. He received his PhD from the University of Illinois at Urbana-Champaign and was a research fellow at the Harvard Medical School, an Alexander von Humboldt Research Fellow at the TU Munich, a member of the Laboratory for Fluorescence Dynamics at the University of Illinois at Urbana-Champaign and a visiting scientist at the University of Ulm. His research centers around ultra-sensitive fluorescence methods, single molecule studies, protein function and dynamics, fluorescence fluctuation spectroscopies, live-cell imaging, single particle tracking and single virus tracing. Jens Michaelis is an Assistant Professor at the LMU Munich. After receiving his PhD in physics in 2000, he spent several years as a postdoc at the University of California, Berkley, focusing on single-molecule studies of molecular motors. His research interests are the molecular mechanisms underlying the biological activity of proteins and mechanical properties of polymer molecules. In 2007 he was awarded the Roemer Prize from the LMU for his habilitation work.
Preface PART I: Single-Particle Imaging and Tracking THREE DIMENSIONAL PARTICLE TRACKING IN A LASER SCANNING FLUORESCENCE MICROSCOPE Introduction Image-Based Single-Particle Tracking Methods Advanced Fluorescence Microscopy Techniques for Single-Particle Tracking Two-Photon Excitation Microscopy 3-D Tracking in Image-Based SPT Approaches 3-D Tracking in Laser Scanning Microscopes Instrumentation Background Noise Simultaneous Two-Particle Tracking Application: Chromatin Dynamics in Interphase Cells Conclusions THE TRACKING OF INDIVIDUAL MOLECULES IN CELLS AND TISSUES Introduction Single-Molecule and Single-Particle Localization Positional Accuracy Tracking Trajectory Analysis Applications Conclusions MESSENGER RNA TRAFFICKING IN LIVING CELLS Intranuclear Structure and Dynamics FCS and FRAP Studies of Nuclear mRNP Mobility Single-Particle Tracking of mRNA Molecules Single-Particle Tracking of Specific, Native mRNPs In Vivo Labeling of Native BR2 mRNPs Outlook: Light Sheet-Based Single-Molecule Microscopy QUANTUM DOTS: INORGANIC FLUORESCENT PROBES FOR SINGLE-MOLECULE TRACKING EXPERIMENTS IN LIVE CELLS Introduction Fluorescent Labels for Single-Molecule Tracking in Cells Optical Properties of Colloidal Quantum Dots Synthesis of Colloidal Fluorescent Quantum Dots Surface Chemistry for the Water-Solubilization of Quantum Dots Interfacing Quantum Dots with Biology Single Quantum Dot Tracking Experiments in Live Cells Conclusions and Perspectives PART II: Energy Transfer on the Nanoscale SINGLE-PAIR FRET: AN OVERVIEW WITH RECENT APPLICATIONS AND FUTURE PERSPECTIVES Introduction Principles of FRET spFRET in Solution spFRET on Immobilized Molecules Future Prospects ALTERNATING-LASER EXCITATION AND PULSED-INTERLEAVED EXCITATION OF SINGLE MOLECULES Introduction ALEX: The Principles of Operation MyALEX Nanosecond-ALEX/Pulsed Interleaved Excitation (PIE) msALEX Three-Color ALEX Conclusions and Outlook UNRAVELING THE DYNAMICS BRDIGING PROTEIN STRUCTURE AND FUNCTION ONE MOLECULE AT A TIME Introduction Converting Chemical Energy to Mechanical Work: Molecular Motors Allostery in Proteins Enzyme Catalysis Conclusions QUANTITATIVE DISTANCE AND POSITION MEASUREMENT USING SINGLE-MOLECULE FRET Introduction Fundamentals of FRET FRET as a Spectroscopic Ruler: Initial Experiments and Limitations Measuring the Quantum Yield The Orientation of Donor and Acceptor Molecules Accurate FRET Measurements Using Fluorescence Correlation Spectroscopy FRET-Based Triangulation and the Nanopositioning System Conclusions and Outlook PART III: Single Molecules in Nanosystems COHERENT AND INCOHERENT COUPLING BETWEEN A SINGLE DIPOLAR EMITTER AND ITS NANOENVIRONMENT Introduction Systems Coupling of Two Oscillating Dipoles A Dipole Close to a Surface A Single Molecule and a Single Nanoparticle Modification of the Spontaneous Emission and Quantum Efficiency by Nanoantennae Conclusions ENERGY TRANSFER IN SINGLE CONJUGATED POLYMER CHAINS Introduction Why Single Chain Spectroscopy? Experimental Approach and Material Systems Photophysics of Single Conjugated Polymer Chains Energy Transfer in Single Chains Influence of Initial Excitation Energy on Energy Transfer Conclusions REACTIONS AT THE SINGLE-MOLECULE LEVEL Introduction Biocatalysis at the Single-Molecule Level Chemocatalysis at the Single-Molecule Level VISUALIZING SINGLE-MOLECULE DIFFUSION IN NANOCHANNEL SYSTEMS Introduction Correlation of Structural and Dynamic Properties Using TEM and SMT Phase Mixture Heterogeneous Dynamics of a Single Molecule Oriented Single Molecules with Switchable Mobility in Long Unidimensional Nanochannels High Localization Accuracy of Single Molecules Down to the Single Channel Limit Probing Chemical Interactions in Silica Thin Films Using Fluorescence Correlation Spectroscopy (FCS) Functionalized Mesoporous Silica Structures Single-Molecule Studies of Mesoporous Silica Structures for Drug-Delivery Applications Conclusions and Outlook