Live Cell Imaging: A laboratory manual (2^nd Ed.)

The second edition of Live Cell Imaging: A Laboratory Manual expands upon and extends the collection of established and evolving methods for studying dynamic changes in living cells and organisms presented in the well-known first edition. There are 16 new chapters and the 21 updated chapters in this new edition. They include advances in atomic force microscopy, structured illumination microscopy and other 3-D approaches, as well as imaging in single cells in animals and in plants. New analytical options include live high-throughput/high-content screening in mammalian cells and computational analysis of live cell data. The manual presents hands-on techniques as well as background material, and can serve as a text in advanced courses. The first section covers principles and fundamental issues of detection and imaging, the second provides detailed protocols for imaging live systems.

Section I: Detection and Approaches to Live Cell Imaging. Fluorescent Protein Tracking and Detection. Constructing and Expressing Fluorescent Protein Fusions. Micropatterning Cell-Substrate Adhesions Using Linear Polyacrylamide as the Blocking Agent. CCD Cameras for Fluorescence Imaging of Living Cells. Fluorescence Perturbation Techniques to Study Mobility and Molecular Dynamics of Proteins in Live Cells: FRAP, Photoactivation, Photoconversion, and FLIP. Imaging Protein State in Cells. A Versatile, Multicolor Total Internal Reflection Fluorescence and Spinning-Disk Confocal Microscope System for High-Resolution Live Cell Imaging. Confocal Microscopy, Deconvolution, and Structured Illumination Methods. Atomic-Force Microscopy for Biological Imaging and Mechanical Testing across Length Scales. OMX: A New Platform for Multimodal, Multichannel Wide-Field Imaging. Digital Scanned Laser Light-Sheet Fluorescence Microscopy. First Step for Fluorescence Correlation Spectroscopy for the Live Cell. Tracking and Quantitative Analysis of Dynamic Movements of Cells and Particles. Imaging Techniques for Measuring Cell Materials Properties. Computational Image Analysis of Cellular Dynamics: A Case Study Based on Particle Tracking. Software Tools, Data Structures and Interfaces for Microscope Imaging. High-Throughput Microscopy Using Live Mammalian Cells. Section II: Imaging of Live Cells and Organisms. In Vivo Imaging of Mammalian Cells. Live Cell Imaging of Yeast. Live Imaging of Caenorhabditis elegans. Live Cell Imaging of Plants.Pushing the Limits of Live Cell Imaging in Drosophila. Dynamic, Long-Term, In Vivo Imaging of Tumor-Stroma Interactions in Mouse Models of Breast Cancer Using Spinning Disk Confocal Microscopy. High-Resolution Multiphoton Imaging of Tumors In Vivo. Correlated Live Cell Light and Electron Microscopy Using Tetracysteine Tags and Biarsenicals. Intravital Microscopy of Normal and Diseased Tissues in the Mouse. Imaging Lipids in Living Cells. Development of Mammalian Cell Lines with lac Operator-Tagged Chromosomes. Imaging Gene Expression in Living Cells. Studying Mitosis in Cultured Mammalian Cells. Imaging Intermediate Filament Proteins in Living Cells. Methods for Expressing and Analyzing GFP-Tubulin and GFP-Microtubule-Associated-Proteins. Imaging of Membrane Systems and Membrane Traffic in Living Cells. Imaging Live Cells under Mechanical Stress. Imaging Single Molecules Using Total Internal Reflection Microscopy. Cellular Imaging Using Total Internal Reflection Fluorescent Microscopy (TIRFM). Visualization and Quantification of Single RNA Molecules in Living Cells.