Modern Biophysical Chemistry: Detection and Analysis of Biomolecules

This updated and up-to-date version of the first edition continues with the really interesting stuff to spice up a standard biophysics and biophysical chemistry course. All relevant methods used in current cutting edge research including such recent developments as super-resolution microscopy and next-generation DNA sequencing techniques, as well as industrial applications, are explained. The text has been developed from a graduate course taught by the author for several years, and by presenting a mix of basic theory and real-life examples, he closes the gap between theory and experiment. The first part, on basic biophysical chemistry, surveys fundamental and spectroscopic techniques as well as biomolecular properties that represent the modern standard and are also the basis for the more sophisticated technologies discussed later in the book. The second part covers the latest bioanalytical techniques such as the mentioned super-resolution and next generation sequencing methods, confocal fluorescence microscopy, light sheet microscopy, two-photon microscopy and ultrafast spectroscopy, single molecule optical, electrical and force measurements, fluorescence correlation spectroscopy, optical tweezers, quantum dots and DNA origami techniques. Both the text and illustrations have been prepared in a clear and accessible style, with extended and updated exercises (and their solutions) accompanying each chapter. Readers with a basic understanding of biochemistry and/or biophysics will quickly gain an overview of cutting edge technology for the biophysical analysis of proteins, nucleic acids and other biomolecules and their interactions. Equally, any student contemplating a career in the chemical, pharmaceutical or bio-industry will greatly benefit from the technological knowledge presented. Questions of differing complexity testing the reader's understanding can be found at the end of each chapter with clearly described solutions available on the Wiley-VCH textbook homepage under: www.wiley-vch.de/textbooks

Table of Contents:
Introduction: What is Biophysical Chemistry? - An Example from Drug Screening PART I: Basic Methods in Biophysical Chemistry BASIC OPTICAL PRINCIPLES Introduction What Does the Electronic Structure of Molecules Look Like? Orbitals, Wave Functions and Bonding Interactions How Does Light Interact with Molecules? Transition Densities and the Transition Dipole Moment Absorption Spectra of Molecules in Liquid Environments. Vibrational Excitation and the Franck-Condon Principle What Happens After Molecules have Absorbed Light? Fluorescence, Nonradiative Transitions and the Triplet State Quantitative Description of all Processes: Quantum Efficiencies, Kinetics of Excited State Populations and the Jablonski Diagram Problems OPTICAL PROPERTIES OF BIOMOLECULES Introduction Experimental Determination of Absorption and Fluorescence Spectra Optical Properties of Proteins and DNA Optical Properties of Important Cofactors BASIC FLUORESCENCE TECHNQUES Introduction Fluorescent Labelling and Linking Techniques Fluorescence Detection Techniques Fluorscence Polarization Anisotropy Forster Resonance Energy Transfer Fluorescence Kinetics Fluorescence Recovery after Photobleaching Biochemiluminescence CHIROPTICAL AND SCATTERING METHODS Chiroptical Methods Light Scattering Vibrational Spectra of Biomolecules MAGNETIC RESONANCE TECHNIQUES Nuclear Magnetic Resonance of Biomolecules Electron Paramagnetic Resonance MASS SPECTROMETRY Introduction MALDI-TOF ESI-MS Structural and Sequence Analysis Using Mass Spectrometry PART II: Advanced Methods in Biophysical Chemistry FLUORESCENCE MICROSCOPY Introduction Conventional Fluorescence Microscopy Total Internal Reflection Fluorescence Microscopy Light-Sheet Microscopy SUPER-RESOLUTION FLUORESCENCE MICROSCOPY Stimulated Emission Depletion (STED) Microscopy Photoactivated Localization Microscopy (PALM) and Stochastic Optical Reconstruction Microscopy (STORM) 3D Super-Resolution Fluorescence Microscopy Imaging of Live Cells Multicolour Super-Resolution Fluorescence Microscopy Structured Illumination Microscopy SOFI Final Comparison SINGLE-BIOMOLECULE TECHNIQUES Introduction Optical Single-Molecule Detection Fluorescence Correlation Spectroscopy Optical Tweezers Atomic Force Microscopy of Biomolecules Patch Clamping ULTRAFAST- AND NONLINEAR SPECTROSCOPY Introduction Nonlinear Microscopy and Spectroscopy Ultrafast Spectroscopy DNA SEQUENCING AND NEXT-GENERATION SEQUENCING METHODS Sanger Method Next-Generation Sequencing Methods SPECIAL TECHNIQUES Introduction Fluorescing Nanoparticles Surface Plasmon Resonance Detection DNA Origami DNA Microarrays Flow Cytometry Fluorescence In Situ Hybridization Microspheres and Nanospheres ASSAY DEVELOPMENT, READERS AND HIGH-THROUGHPUT SCREENING Introduction Assay Development and Assay Quality Microtitre Plates and Fluorescence Readers Application Example: Drug Discovery and High-Throughput Screening Index