Single Particle Tracking and Single Molecule Energy Transfer

by Brä; uchle, Christoph; Lamb, Don Carroll; Michaelis, Jens

• ISBN13: 9783527322961
• ISBN10: 3527322965
• Edition: 1st
• Format: Hardcover
• Copyright: 2009-12-21
• Publisher: Wiley-VCH

Summary

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.

Author Biography

Christoph Braeuchle is a professor at the Ludwig-Maximilian-University (LMU) Munich. After his PhD, he spent one year as a postdoc at IBM in San Jos+¬, California, USA. His research focuses on imaging, spectroscopy and manipulation of single molecules in bio- and nano-sciences. Beside numerous publications in international journals, Professor Braeuchle has won several honors, including the famous Philip Morris Research award and the Karl Heinz Beckurts Preis 2002.

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 in the Prof Bustamante's group. His research interests are the molecular mechanisms underlying the biological activity of proteins and mechanical properties of polymer molecules.

Don Lamp is Group Leader at the LMU Munich. He received his PhD from the University of Illinois and was a research fellow at the Harvard Medical School and an Alexander von Humboldt Research Fellow at the TU Munich. 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. Dr. Lamp won the 2006 Roemer Prize.

Table of Contents

Preface

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

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

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

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