Imaging and Spectroscopic Analysis of Living Cells - 1st Edition - ISBN: 9780123918574, 9780123918697

Imaging and Spectroscopic Analysis of Living Cells, Volume 504

1st Edition

Optical and Spectroscopic Techniques

eBook ISBN: 9780123918697
Hardcover ISBN: 9780123918574
Imprint: Academic Press
Published Date: 24th January 2012
Page Count: 560
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Table of Contents

Series Page


Volume in series

Laser-Induced Radiation Microbeam Technology and Simultaneous Real-Time Fluorescence Imaging in Live Cells

1. Introduction

2. Experimental Procedure

3. Conclusion/Forward Look


A Cell Biologist's Guide to High Resolution Imaging

1. Introduction

2. Physical Limitations on the Resolution of Conventional Microscopy

3. Preparations for High Resolution Fluorescence Imaging

4. High Resolution Image Data Acquisition and Processing

5. Processing

6. Analysis of High Resolution Image Data

7. Conclusions


Applications of Fluorescence Lifetime Spectroscopy and Imaging to Lipid Domains In Vivo

1. The Challenge of Studying Lipid Domains In Vivo Defied by Fluorescence Lifetimes

2. Importance of Studies In Vitro to Design and Rationalize Studies In Vivo

3. Labeling Cell Membranes In Vivo: Lipophilic Probes Versus GFP-Tagged Membrane Proteins

4. Cuvette Lifetimes and FLIM: In Vivo Applications


Detecting and Tracking Nonfluorescent Nanoparticle Probes in Live Cells

1. Introduction

2. Techniques and Tools

3. Biological Applications

4. Cytotoxicity of Nanoparticle Probes

5. Conclusions and Future Perspective


Fluorescence Lifetime Microscopy of Tumor Cell Invasion, Drug Delivery, and Cytotoxicity

1. Introduction

2. Preparation of Mouse Mammary Tumor Cell Spheroids in a Collagen Matrix

3. Simultaneous Acquisition of Fluorescence Lifetime and Intensity to Monitor DOX Uptake

4. Monitoring Drug Uptake Kinetics by FLIM

5. Conclusions and Outlook


Measuring Membrane Protein Dynamics in Neurons Using Fluorescence Recovery after Photobleach

1. Introduction

2. Experimental Parameters and Preparation

3. Equipment

4. Establishing FRAP Conditions

5. A Basic FRAP Protocol

6. Analysis of FRAP


Fluorescent Speckle Microscopy in Cultured Cells

1. Introduction

2. Technical Challenges of FSM

3. Detailed Methodology for FSM in Cultured Cells

4. Future Challenges


Green-to-Red Photoconvertible mEosFP-Aided Live Imaging in Plants

1. Introduction

2. Expression of mEosFP Fusion Proteins in Plants

3. Visualization of mEosFP Probes in Plants

4. Uses of mEosFP Probes in Plants

5. Post Acquisition Image Processing and Data Creation


Methods for Cell and Particle Tracking

1. Introduction

2. Tracking Approaches

3. Tracking Tools

4. Tracking Measures

5. Tips and Tricks


Correlative Light-Electron Microscopy

1. Introduction

2. Observation of Living Cells and Fixation

3. Immunolabeling with NANOGOLD

4. Enhancement of Sample Contrast, Sample Locating, and Embedding

5. Identification of the Cell of Interest on EPON Blocks

6. Sample Orientation and EM Sectioning from the Very First Section

7. Picking up Serial Sections with the Empty Slot Grid

8. EM Analysis


Optical Techniques for Imaging Membrane Domains in Live Cells (Live-Cell Palm of Protein Clustering)

1. Introduction

2. Sample Preparation and Data Acquisition

3. Data Analysis


Single Live Cell Topography and Activity Imaging with the Shear-Force-Based Constant-Distance Scanning Electrochemical Microscope

1. Introduction

2. Shear-Force-Based Constant-Distance Scanning Electrochemical Microscopy for Live Cell Studies: Apparatus, Probes, and Operation

3. Selected Applications of SF-CD-SECM Live Cell Studies


Visualization of TGN-Endosome Trafficking in Mammalian and Drosophila Cells

1. Introduction

2. Molecular Tools

3. Live-Cell Imaging in Mammalian Cells

4. Live-Cell Imaging in Drosophila Cells

5. Conclusion Remarks

Live Cell Imaging with Chemical Specificity Using Dual Frequency CARS Microscopy

1. Introduction

2. “Noninvasive” Live Cell Imaging

3. Experimental Setup

4. Maximizing Collection Efficiency for Live Cell Imaging


Imaging Intracellular Protein Dynamics by Spinning Disk Confocal Microscopy

1. Introduction

2. Instrument Design

3. Combination with Other Imaging Techniques

4. Specimen Preparation


Visualizing Dynamic Activities of Signaling Enzymes Using Genetically Encodable Fret-Based Biosensors

1. Introduction

2. Generalizable Modular Designs

3. FRET-Based Biosensors for Monitoring Signaling Enzymes

4. Example: A-kinase Activity Reporter (AKAR)

5. Summary and Perspectives


Live-Cell Imaging of Aquaporin-4 Supramolecular Assembly and Diffusion

1. Aquaporin-4 (AQP4) and Orthogonal Arrays of Particles

2. Approaches to Image AQP4 and OAPs

3. AQP4 Diffusion and OAPs Studied by Quantum Dot Single Particle Tracking

4. OAP Dynamics and Structure Studied with GFP-AQP4 Chimeras

5. Single-Molecule Analysis Shows AQP4 Heterotetramers

6. Photobleaching Reveals Post-Golgi Assembly of OAPs

7. Super-Resolution Imaging of AQP4 OAPs

Coiled-Coil Tag–Probe Labeling Methods for Live-Cell Imaging of Membrane Receptors

1. Introduction

2. Various Principles Used for Tag–Probe Labeling

3. Coiled-Coil Tag–Probe Labeling

4. Applications


Monitoring Protein Interactions in Living Cells with Fluorescence Lifetime Imaging Microscopy

1. Introduction

2. FD FLIM Measurements

3. Measuring Protein–Protein Interactions in Living Cells Using FLIM–FRET

4. The Strengths and Limitations of FLIM


Open Source Tools for Fluorescent Imaging

1. Why Open Source Software?

2. Open Source Software for Microscopy Imaging and Analysis

3. The Future



This volume of Methods in Enzymology is the first of three parts looking at current methodology for the imaging and spectroscopic analysis of live cells. The chapters provide hints and tricks not available in primary research publications. It is an invaluable resource for academics, researchers and students alike.

Key Features

  • Expert authors who are leaders in the field
  • Extensively referenced and useful figures and tables
  • Provides hints and tricks to facilitate reproduction of methods


Biochemists, biophysicists, molecular biologists, analytical chemists, pharmacologists and physiologists


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