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Beschreibung 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 Biology? 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.
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Inhalt Part I: General Principles and Overview
1. Instrumentation for Live-Cell Imaging and Main Formats
Martin Oheim
2. Labels and Probes for Live Cell Imaging: Overview and Selection Guide
Scott Hilderbrand
3. Live Cell Imaging: An Industrial Perspective
Terry McCann
Part II: Imaging Techniques, Probes, and Applications
4. Design of Fluorescent Fusion Protein Probes
Elizabeth Pham and Kevin Truong
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 Shen, Yan Chen, and Paul Kwong-Hang Tam
6. Photo-Activatable Probes for the Analysis of Receptor Function in Living Cells
Wen-hong Li
7. The Application of Fluorescent Probes for the Analysis of Lipid Dynamics During Phagocytosis
Ronald S. Flannagan and Sergio Grinstein
8. Imaging of Mitotic Cell Division and Apoptotic Intra-Nuclear Processes in Multicolor
Kenji Sugimoto and Shigenobu Tone
9. Manipulation of Neutrophil-Like HL-60 Cells for the Study of Directed Cell Migration
Arthur Millius and Orion D. Weiner
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, and Susan K. Pierce
11. Sample Preparation for STED Microscopy
Christian A. Wurm, Daniel Neumann, Roman Schmidt, Alexander Egner, and Stefan Jakobs
12. Two-Photon Permeabilization and Calcium Measurements in Cellular Organelles
Oleg Gerasimenko and Julia Gerasimenko
13. Imaging and Analysis of Three-Dimensional Cell Culture Models
Benedikt W. Graf and Stephen A. Boppart
14. Long-Term Imaging in Microfluidic Devices
Gilles Charvin, Catherine Oikonomou, and Frederick Cross
15. Monitoring of Cellular Responses to Hypoxia
Christoph Wotzlaw and Joachim Fandrey
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, and Dmitri B. Papkovsky
17. Analysis of Mitochondrial pH and Ion Concentrations
Martin vandeVen, Corina Balut, Szilvia Baron, Ilse Smets, Paul Steels, and Marcel Ameloot
18. Live Cell Imaging Analysis of Receptor Function
Daniel C. Worth and Maddy Parsons
19. Subcellular Dynamic Imaging of Protein-Protein Interactions in Live Cells by Bioluminescence Resonance Energy Transfer
Julie Perroy
20. Quantitative Analysis of Membrane Potentials
Manus W. Ward
21. Image Correlation Spectroscopy to Define Membrane Dynamics
Jeremy Bonor and Anja Nohe
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