Modern Biophysical Chemistry. Detection and Analysis of Biomolecules. 2nd Edition

  • ID: 2755171
  • Book
  • 360 Pages
  • John Wiley and Sons Ltd
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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: [external URL]

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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

Förster 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

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Peter Jomo Walla
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