Live Cell Imaging: Methods and Protocols / Edition 1

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Now a routine tool in biomedical and life science research, live cell imaging has made major progress enabling this core biochemical, cell, and molecular biology technique to become even more powerful, versatile, and affordable. In Live Cell Imaging: Methods and Protocols, a panel of expert contributors provide a comprehensive compendium of experimental approaches to live cell imaging in the form of several overview chapters followed by representative examples and case studies covering different aspects of the most current methodology. By examining a range of state-of-the-art protocols extensively validated in complex biological studies, this volume highlights new experimental and instrumental opportunities and helps researchers to select appropriate imaging methods for their specific biological questions and measurement tasks. Written in the highly successful Methods in Molecular BiologyT series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Live Cell Imaging: Methods and Protocols promises to contribute greatly to the further development and dissemination of this fundamentally important technology which spans across many disciplines including molecular and cell biology, chemistry, physics, optics, engineering, cell physiology, and medicine. Written for: Molecular and cellular biologists, chemists, physicists, optics specialists, engineers, cell physiologists, and medical doctors

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Editorial Reviews

Doody's Review Service
Reviewer: Bruce A. Fenderson, PhD (Thomas Jefferson University)
Description: This book describes innovative approaches for analyzing cellular organelles and biomolecules in real time. A volume in the popular Methods in Molecular Biology series, it provides a compendium of experimental approaches to live cell imaging in the form of overview chapters followed by representative examples and case studies. The 21 chapters cover fascinating topics ranging from design of fluorescent protein probes to monitoring cellular responses to hypoxia. The emphasis is on cell biology, cell physiology, and biophysics.
Purpose: According to the editor, this book "provides a range of state-of-the-art protocols extensively validated in complex biological studies." The editor hopes that this volume will contribute to the further development and dissemination of...fundamentally important technology which spans across many disciplines.
Audience: The book is written for graduate students, postdoctoral fellows, and research scientists interested in molecular and cellular biology, biochemistry, and biophysics. It will also be appreciated by basic science and clinical researchers interested in physics, optics, engineering, cell physiology. The authors are internationally recognized experts in this growing field.
Features: The book is divided into two major sections: principles and techniques/applications. Some of the chapters present scholarly reviews, while others narrowly focus on specific research protocols. Each chapter includes an abstract, key words, introduction, research protocols, notes for trouble-shooting, and references to facilitate further research. The overview section of the book helps place the various chapters into context. The tables, chemical formulas, figures, and line drawings are clear and informative. Some of the photomicrographs are in color. The depth and breadth of topics the book covers is impressive.
Assessment: The advantage of this book is that it brings together a wide variety of approaches to live cell imaging and provides exciting examples of current research findings. The authors carefully explain theory and practice with straightforward, step-by-step protocols. One minor criticism is that the experimental approaches described in this book are complicated, and many require special equipment. However, the editor believes that "live cell imaging has now become a routine tool in biomedical and life science research." The equipment required is gradually moving from core facilities to small laboratories. The authors focus on innovative topics and provide a snapshot of application. This will be an excellent resource for basic science researchers interested in real-time analysis of organelles and biomolecules in living cells and tissues.
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Product Details

  • ISBN-13: 9781607614036
  • Publisher: Springer-Verlag New York, LLC
  • Publication date: 12/17/2009
  • Series: Methods in Molecular Biology Series, #591
  • Edition description: 2010
  • Edition number: 1
  • Pages: 367
  • Product dimensions: 7.20 (w) x 10.10 (h) x 1.20 (d)

Table of Contents

Preface v

Contributors ix

Part I General Principles and Overview 1

1 Instrumentation for Live-Cell Imaging and Main Formats Martin Oheim 3

2 Labels and Probes for Live Cell Imaging: Overview and Selection Guide Scott A. Hilderbrand 17

3 Live Cell Imaging: An Industrial Perspective Terry McCann 47

Part II Imaging Techniques, Probes, and Applications 67

4 Design of Fluorescent Fusion Protein Probes Elizabeth Pham Kevin Truong 69

5 Synthetic Fluorescent Probes for Imaging of Peroxynitrite and Hypochlorous Acid in Living Cells Dan Yang Zhen-Ning Sun Tao Peng Hua-Li Wang Jian-Gang Sben Yan Chen Paul Kwong-Hang Tarn 93

6 Photo-Activatable Probes for the Analysis of Receptor Function in Living Cells Wen-Hong Li 105

7 The Application of Fluorescent Probes for the Analysis of Lipid Dynamics During Phagocytosis Ronald S. Flannagan Sergio Grinstein 121

8 Imaging of Mitotic Cell Division and Apoptotic Intra-Nuclear Processes in Multicolor Kenji Sugimoto Shigenobu Tone 135

9 Manipulation of Neutrophil-Like HL-60 Cells for the Study of Directed Ceil Migration Arthur Millius Orion D. Weiner 147

10 A Method for Analyzing Protein-Protein Interactions in the Plasma Membrane of Live B Cells by Fluorescence Resonance Energy Transfer Imaging as Acquired by Total Internal Reflection Fluorescence Microscopy Hae Won Sohn Pavel Tolar Joseph Brzostowski Susan K. Pierce 159

11 Sample Preparation for STED Microscopy Christian A. Wurm Daniel Neumann Roman Schmidt Alexander Egner Stefan Jakobs 185

12 Two-Photon Permeabilization and Calcium Measurements in Cellular Organelles Oleg Gerasimenko Julia Gerasimenko 201

13 Imaging and Analysis of Three-Dimensional Cell Culture Models Benedikt W. Graf Stephen A. Boppart 211

14 Long-Term Imaging in Microfluidic Devices Gilles Charvin Catherine Oikonomou Frederick Cross 229

15 Monitoring of Cellular Responses to Hypoxia Christoph Wotzlaw Joachim Fandrey 243

16 Imaging of Cellular Oxygen and Analysis of Metabolic Responses of Mammalian Cells Andreas Fercher Tomas C. O'Riordan Alexander V. Zhdanov Ruslan I. Dmitriev Dmitri B. Papkovsky 257

17 Analysis of Mitochondrial pH and Ion Concentrations Martin vandeVen Corina Balut Szilvia Baron Ilse Smets Paul Steels Marcel Ameloot 275

18 Live Cell Imaging Analysis of Receptor Function Daniel C. Worth Maddy Parsons 311

19 Subcellular Dynamic Imaging of Protein-Protein Interactions in Live Cells by Biohiminescence Resonance Energy Transfer Julie Perroy 325

20 Quantitative Analysis of Membrane Potentials Mmius W. Ward 335

21 Image Correlation Spectroscopy to Define Membrane Dynamics Jeremy Bonor Anja Nohe 353

Subject Index 365

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