Overview

Increasing numbers of physicists, chemists, and mathematicians are moving into biology, reading literature across disciplines, and mastering novel biochemical concepts. To succeed in this transition, researchers must understand on a practical level what is experimentally feasible. The number of experimental techniques in biology is vast and often specific to particular subject areas; nonetheless, there are a few basic methods that provide a conceptual underpinning for broad application. Introduction to Experimental Biophysics is the ideal benchtop companion for physical scientists interested in getting their hands wet. Assuming familiarity with basic physics and the scientific method but no previous background in biology or chemistry, this book provides: A thorough description of modern experimental and analytical techniques used in biological and biophysical research Practical information and step-by-step guidance on instrumentation and experimental design Recipes for common solutions and media, lists of important reagents, and a glossary of biological terms used Developed for graduate students in biomedical engineering, physics, chemical engineering, chemistry, mathematics, and computer science, Introduction to Experimental Biophysics is an essential resource for scientists to overcoming conceptual and technical barriers to working in a biology wet lab.

Full Product Details

Author:   Jay L. Nadeau (Portland State University, OR, USA)
Publisher:   Taylor & Francis Inc
Imprint:   CRC Press Inc
Dimensions:   Width: 17.80cm , Height: 3.30cm , Length: 25.40cm
Weight:   1.202kg
ISBN:  

9781439829530

ISBN 10:   1439829535
Pages:   672
Publication Date:   21 September 2011
Audience:   College/higher education ,  General/trade ,  Tertiary & Higher Education ,  General
Replaced By:   9781498799591
Format:   Paperback
Publisher's Status:   Unknown
Availability:   Awaiting stock  

Table of Contents

Introduction and Background Basic Biochemistry Energies and Potentials Principles of Spectroscopy Cells DNA, RNA, Replication, and Transcription Translation and the Genetic Code Protein Folding and Trafficking Alternative Genetics What Is Cloning? Design of a Molecular Biology Experiment and How to Use This Book Questions and Problems Background Reading Molecular Cloning of DNA and RNA Introduction Obtaining and Storing Plasmids Selection of an Appropriate E. coli Amplification Strain: Transformation of E. coli with Plasmid Plasmid Amplification and Purification Plasmid Restriction Mapping and Agarose Gel Electrophoresis An Example of Cloning Experiment Cloning by the Polymerase Chain Reaction Sequencing RNA Methods Southern and Northern Blots Solutions for Large Cloning Problems and Multiple Inserts Mutagenesis and Directed Evolution Microarrays Summary Questions and Problems Background Reading Expression of Genes in Bacteria, Yeast, and Cultured Mammalian Cells Introduction Expressing Genes in Microorganisms Mammalian Cell Culture Transfection of Mammalian Cells I: Standard Techniques Transfection of Mammalian Cells II: Specialized Physical Methods for Special Occasions Transfection of Mammalian Cells III: Viruses Summary Questions and Problems Background Reading Protein Expression Methods Introduction Expression Systems Identification of a DNA Source Selecting an Expression Vector Subcloning into an Expression Vector Selection of an Expression Strain or Cell Line Protein Expression Checking Protein Expression (and Purity) Using SDS-PAGE Protein Isolation and Purification Chromatography Buffer Exchange and Concentration Example Experiment: Expression and Purification of Fluorescent Protein Dronpa Conclusions and Final Remarks Background Reading Protein Crystallization Introduction Crystallization of Macromolecules Preparation of Proteins for Crystallization Components of Crystallization Solutions Other Factors Affecting Crystallization Crystallization Strategies Example Experiment: Lysozyme Data Collection and Structure Determination Using X-Ray Crystallography A Special Case: Membrane Proteins Troubleshooting Q&A Conclusions and Final Remarks Questions and Problems Background Reading Introduction to Biological Light Microscopy Introduction The Physics of Microscopy: Magnification and Resolution Anatomy of a Biological Microscope Brightfield Imaging Techniques Basic Fluorescence Microscopy Fluorophores for Cell Labeling Fluorescent Proteins Multispectral Imaging Using Acousto-Optical Tunable Filters Advanced Techniques Summary and Remarks Questions and Problems Background Reading Quantitative Cell Culture Techniques Introduction Quantifying Bacterial Growth and Death Quantifying Mammalian Cells Flow Cytometry Example Experiment: Determining Leukemic B-Cells and T-Cells by Flow Cytometry Quantifying Viruses Measuring Cell Populations Using Quantitative PCR Summary and Final Remarks Questions and Problems Background Reading Semiconductor Nanoparticles (Quantum Dots) Introduction Quantum Dot Properties and Synthesis QD Applications Example Experiment: Conjugation of Quantum Dots to Dopamine and Quantifying the Effects on Fluorescence per Molecule Bound Summary and Remarks Questions and Problems Background Reading Gold Nanoparticles Introduction The Physics of Scattering and Spherical Metal Nanoparticles Synthesis of Gold Nanoparticles Characterization and Surface Modification of Gold Nanoparticles Applications for Colorimetric Detection and Microscopy Sample Experiment: Labeling Cells with Lectin-Tagged Au Nanoparticles Applications in Surface-Enhanced Raman Scattering Gold Nanoparticles as Photothermal Transducers Conclusion Questions and Problems Background Reading Surface Functionalization Techniques Introduction Preparing Monolayers Using Functional Silanes or Thiols Techniques for Characterizing Surface Monolayers Functionalization of Modified Surfaces Using Cross-Linkers Example Experiment: Preparing a Silane–Biotin–Streptavidin Sandwich on SiO2 Features on an Si Chip Preventing Nonspecific Binding of Biomolecules Assembling Membrane Proteins on Surfaces Testing the Function of Immobilized Proteins Conclusion and Final Remarks Questions and Problems Background Reading Electrophysiology Introduction Physical Basis and Circuit Models Solutions and Blockers Instrumentation Lipid Bilayer Setup Cell Patch-Clamp Setup: What Is Needed? The Art and Magic of Pipette Pulling Step-by-Step Guide to Perform a Whole-Cell Recording Example Experiment: Whole-Cell Recording on Cells A Brief Introduction to Single-Channel Modeling and Data Analysis Network Recording Conclusions and Final Remarks Questions and Problems Background Reading Spectroscopy Tools and Techniques Introduction Guiding Principles UV–Vis Absorbance Spectroscopy Fluorescence Spectroscopy Time-Resolved Emission Time-Resolved Absorption Infrared Spectroscopy Nuclear Magnetic Resonance Electron Paramagnetic Resonance Spectroscopy X-Ray Spectroscopy Example Experiment: Characterization of CdSe/ZnS Nanoparticle Bioconjugate Using UV–Vis, Fluorescence Emission, Time-Resolved Emission, FTIR, and EPR Spectroscopy Final Comments Questions and Problems Background Reading Appendix Glossary Index

Reviews

This book fills the need for a practical, hands-on guide for physical scientists who are moving into biological research. --Professor Daniel A. Beard, Medical College of Wisconsin As scientists from more quantitative fields expand further into molecular and cellular biology, their labs need to acquire new biological methods for sample preparation and handling. These skills are not traditionally available to physicists and chemists. This book will be appropriate for any experimentalist in chemistry or physics who is moving into biological work. It will also be excellent reading material for undergraduate or graduate students who will be working in a biologically oriented lab, as well as for an advanced lab class in biophysics or bioengineering. --Professor Mark C. Williams, Northeastern University This book will be very useful for training the growing number of researchers and students from physical sciences to become more familiar with techniques used in biology. The author has made a great effort to keep everything defined and simple. --Professor James A. Forrest, Department of Physics and Associate Dean of Research, Faculty of Science, University of Waterloo

Author Information

Jay L. Nadeau is an associate professor of biomedical engineering and physics at McGill University (2004--present). Her research interests include nanoparticles, fluorescence imaging, and development of instrumentation for the detection of life elsewhere in the solar system. She has published over 50 papers on topics ranging from theoretical condensed matter physics to experimental neurobiology to the development of anticancer drugs and, in the process, has used almost every technique described in this book. Her work has been featured in New Scientist, Highlights in Chemical Biology, Radio Canada's Les Annees Lumiere, Le Guide des Tendances, and in educational displays in schools and museums. Her research group features chemists, microbiologists, roboticists, physicists, and physician-scientists, all learning from each other and hoping to speak each other's language. A believer in bringing biology to physicists as well as physics to biologists, she has created two graduate-level courses: methods in molecular biology for physical scientists and mathematical cellular physiology. She also teaches pharmacology in the medical school and was one of the pioneers in the establishment of multiple mini-interviews for medical school admission. She has an adjunct position with The Jackson Laboratory in Bar Harbor, Maine, and collaborators in industry and academia in the United States, Europe, Australia, and Japan. She has given several dozen invited talks at meetings of the American Chemical Society, American Geophysical Union, the International Society for Optics and Photonics (SPIE), the Committee on Space Research, and many others. Before McGill, she was a member of the Jet Propulsion Laboratory's Center for Life Detection, and previous to that a Burroughs-Wellcome postdoctoral scholar in the laboratory of Henry A. Lester at Caltech. She received her PhD in physics from the University of Minnesota in 1996.

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