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Current Protocols Select : Methods and Appplications in Microscopy and Imaging,9781118044315
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Current Protocols Select : Methods and Appplications in Microscopy and Imaging

by ;
Edition:
1st
ISBN13:

9781118044315

ISBN10:
1118044312
Media:
Paperback
Pub. Date:
9/3/2013
Publisher(s):
Current Protocols
List Price: $149.95

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Summary

This book is intended to be a resource for those new to the use of the microscope as well as for seasoned investigators, exposing them to the wealth of opportunities to visualize the molecules and cellular processes they are studying. Because these are Current Protocols articles, they provide detailed step-by-step instructions for the various methods and applications, described and written at a level that lets every investigator employ even very sophisticated microscopy methods. The first part of the book will focus on the fundamentals of microscopy and its function, followed by descriptions of the various technologies. Subsequently, the body of the text will be composed of a series of chapters giving application-specific examples with recipes and recommendations for each approach.

Table of Contents

Chapter 1 Fundamentals of the Microscope
Introduction
Fluorescence Microscopy: A Concise Guide to Current Imaging Methods
Microscope Objectives 
Light Microscopy Digital Imaging
Optical Filtering Systems for Wavelength Selection in Light Microscopy 
Proper Adjustment and Alignment of the Light Microscope 
Chapter 2 Basic Methods
Introduction
Section I. Sample Preparation for Conventional Microscopy
Cryosectioning 
Immunohistochemistry 
Section II. Dyes and Probes
Overview on new compartment probes for imaging in cell biology
Fluorescent Protein Color Palette
Photoactivation and Imaging of Photoactivatable Fluorescent Proteins
Section III. Optical Sectioning Microscopy
Basic Confocal Microscopy 
Evaluation and Purchase of confocal microscopes: Some of the Numerous Factors to Consider
3D Deconvolution Microscopy 
Multiphoton Imaging 
Chapter 3 Applications
Introduction
Section I. Basic Live Cell Imaging
Building a Live Cell Microscope
Time-Lapse Microscopy Approaches to Track Cell Cycle Progression at the Single-Cell 
Analysis of Mitochondrial Dynamics and Functions Using Imaging Approaches 
Fluorescence Recovery After Photobleaching (FRAP) 
Confocal Imaging of Cell Division
Total Internal Reflection (TIRF) Microscopy 
Section II. Fluorescence Resonance Energy Transfer
Imaging Protein-Protein Interactions by ForterResonance Energy Transfer (FRET) Microscopy  in live cells
Imaging Protein-Protein Interactions in Fluorescence Resonance Energy Transfer (FRET) Microscopy 
Use of Spectral Fluorescence Resonance Energy Transfer to Detect Nitric Oxide-Based Signaling Events in Isolated Perfused Lung 
Section III. Imaging of Model Systems
Fluorescent Imaging Techniques for Studying Drosophila Embryo Development 
Time-Lapse Imaging of Embryonic Neural Stem Cell Division in Drosophila by Two-Photon Microscopy
Imaging Tumor Cell Movement in Vivo
Live Animal Imaging of Renal Function by Multiphoton Microscopy 
Biological Second and Third Harmonic Microscopy 
2P Imaging of the Immune System
Section IV. Super-Resolution Methods
Super-resolution Microscopy: A Comparative Treatment
Photoactivated Localization Microscopy (PALM) of Adhesion Complexes 
Comparative and Practical Aspects of Localization-Based Super Resolution Imaging
Chapter 4 Image Analysis
Introduction
Ethical Considerations When Altering Digital Images 
From Image to Data Using Common Image-Processing Techniques
Practical Considerations When Altering Digital Images
 
Appendix 1: Common Buffers and Reagents
Chapter 1 Fundamentals of the Microscope
Introduction
  • Fluorescence Microscopy: A Concise Guide to Current Imaging Methods
  • Microscope Objectives 
  • Light Microscopy Digital Imaging
  • Optical Filtering Systems for Wavelength Selection in Light Microscopy 
  • Proper Adjustment and Alignment of the Light Microscope 

Chapter 2 Basic Methods
Introduction
Section I. Sample Preparation for Conventional Microscopy
  • Cryosectioning 
  • Immunohistochemistry 
Section II. Dyes and Probes
  • Overview on new compartment probes for imaging in cell biology
  • Fluorescent Protein Color Palette
  • Photoactivation and Imaging of Photoactivatable Fluorescent Proteins
Section III. Optical Sectioning Microscopy
  • Basic Confocal Microscopy 
  • Evaluation and Purchase of confocal microscopes: Some of the Numerous Factors to Consider
  • 3D Deconvolution Microscopy 
  • Multiphoton Imaging
 
Chapter 3 Applications
Introduction
Section I. Basic Live Cell Imaging
  • Building a Live Cell Microscope
  • Time-Lapse Microscopy Approaches to Track Cell Cycle Progression at the Single-Cell 
  • Analysis of Mitochondrial Dynamics and Functions Using Imaging Approaches 
  • Fluorescence Recovery After Photobleaching (FRAP) 
  • Confocal Imaging of Cell Division
  • Total Internal Reflection (TIRF) Microscopy 
Section II. Fluorescence Resonance Energy Transfer
  • Imaging Protein-Protein Interactions by ForterResonance Energy Transfer (FRET) Microscopy  in live cells
  • Imaging Protein-Protein Interactions in Fluorescence Resonance Energy Transfer (FRET) Microscopy 
  • Use of Spectral Fluorescence Resonance Energy Transfer to Detect Nitric Oxide-Based Signaling Events in Isolated Perfused Lung 
Section III. Imaging of Model Systems
  • Fluorescent Imaging Techniques for Studying Drosophila Embryo Development 
  • Time-Lapse Imaging of Embryonic Neural Stem Cell Division in Drosophila by Two-Photon Microscopy
  • Imaging Tumor Cell Movement in Vivo
  • Live Animal Imaging of Renal Function by Multiphoton Microscopy 
  • Biological Second and Third Harmonic Microscopy 
  • 2P Imaging of the Immune System
Section IV. Super-Resolution Methods
  • Super-resolution Microscopy: A Comparative Treatment
  • Photoactivated Localization Microscopy (PALM) of Adhesion Complexes 
  • Comparative and Practical Aspects of Localization-Based Super Resolution Imaging

Chapter 4 Image Analysis
Introduction
  • Ethical Considerations When Altering Digital Images 
  • From Image to Data Using Common Image-Processing Techniques
  • Practical Considerations When Altering Digital Images
 
Appendix 1: Common Buffers and Reagents


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