Biomedical Optical Phase Microscopy and Nanoscopy - 1st Edition - ISBN: 9780124158719, 9780124158863

Biomedical Optical Phase Microscopy and Nanoscopy

1st Edition

Editors: Natan T. Shaked Zeev Zalevsky Lisa Satterwhite
eBook ISBN: 9780124158863
Hardcover ISBN: 9780124158719
Imprint: Academic Press
Published Date: 5th November 2012
Page Count: 432
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Written by leading optical phase microscopy experts, this book is a comprehensive reference to phase microscopy and nanoscopy techniques for biomedical applications, including differential interference contrast (DIC) microscopy, phase contrast microscopy, digital holographic microscopy, optical coherence tomography, tomographic phase microscopy, spectral-domain phase detection, and nanoparticle usage for phase nanoscopy

The Editors show biomedical and optical engineers how to use phase microscopy for visualizing unstained specimens, and support the theoretical coverage with applied content and examples on designing systems and interpreting results in bio- and nanoscience applications.

Key Features

  • Provides a comprehensive overview of the principles and techniques of optical phase microscopy and nanoscopy with biomedical applications.
  • Tips/advice on building systems and working with advanced imaging biomedical techniques, including  interpretation of phase images, and techniques for quantitative analysis based on phase microscopy.
  • Interdisciplinary approach that combines optical engineering, nanotechnology, biology and medical aspects of this topic. Each chapter includes practical implementations and worked examples.


Biomedical engineers and researchers
Nanotechnology engineers and researchers
Optical engineers and researchers

Microscopy engineers

Table of Contents


Part 1: Phase Contrast Microscopy and Differential Interference Contrast Microscopy

Chapter 1. Phase Contrast Microscopy

1.1 Introduction

1.2 How Phase Contrast Works

1.3 How to Acquire Phase Contrast Images

1.4 Experimental Uses

1.5 Limitations of Phase Optics


Further Reading

Chapter 2. Differential Interference Microscopy

2.1 Introduction

2.2 Measuring Shear Angle of DIC Prism

2.3 Bias Optimization

2.4 A Quantitative OI-DIC Microscope with Fast Modulation of Bias and Shear Direction

2.5 Combination of OI-DIC and Orientation-Independent Polarization Imaging

2.6 Combination of OI-DIC and Fluorescence Imaging



Chapter 3. Long-Term Recordings of Live Human Cells Using Phase Contrast Microscopy

3.1 Introduction

3.2 The Microscope

3.3 Correlative Live Cell–Fixed Cell Observations


Further Reading

Chapter 4. Phase Imaging in Plant Cells and Tissues

4.1 Out of Focus Imaging

4.2 Dark Field

4.3 Phase Contrast Techniques

4.4 Becke Line in Optical Microscopy

4.5 DIC Microscopy

4.6 Hoffmann Modulation Contrast

4.7 Adaptive Optics Microscopy

4.8 Interferometric Microscopy

4.9 Second Harmonic Imaging Microscopy

4.10 Magnetic Resonance Imaging of Plants Showing the Flow Component



Part 2: Digital Holographic Phase Microscopy

Chapter 5. Digital Holographic Microscopy for Measuring Biophysical Parameters of Living Cells

5.1 Introduction

5.2 Technical Introduction

5.3 Biological Applications



Chapter 6. Holographic Microscopy Techniques for Multifocus Phase Imaging of Living Cells

6.1 Introduction

6.2 DHM Setups for Live Cell Imaging

6.3 Recording and Numerical Evaluation of Digital Holograms

6.4 Refractive Index Determination of Cells in Suspension

6.5 DHM Analysis of Morphology and Thickness Changes

6.6 Quantitative Cell Division Monitoring

6.7 Cell Tracking in 3D Environments

6.8 Conclusions



Chapter 7. Phase Unwrapping Problems in Digital Holographic Microscopy of Biological Cells

7.1 Introduction

7.2 Experimental Techniques

7.3 Varying Reconstruction Distance Method

7.4 Dual-Wavelength Phase Imaging

7.5 Conclusions


Chapter 8. On-Chip Holographic Microscopy and its Application for Automated Semen Analysis

8.1 Introduction

8.2 Partially Coherent In-Line Holography on a Chip

8.3 Reconstruction of Microscopic Images and the Phase Recovery Process

8.4 On-Chip Imaging of Sperm Using Partially Coherent Lensfree In-Line Holography

8.5 High-Throughput On-Chip Semen Analysis Results

8.6 Conclusions



Chapter 9. Synthetic Aperture Lensless Digital Holographic Microscopy (SALDHM) for Superresolved Biological Imaging

9.1 Introduction

9.2 SALDHM Inside the Gabor’s Regime

9.3 SALDHM Outside the Gabor’s Regime

9.4 Conclusions


Chapter 10. Combining Digital Holographic Microscopy with Microfluidics: A New Tool in Biotechnology

10.1 Introduction

10.2 Drive and Analyze

10.3 Particle Tracking in 3D

10.4 Conclusions


Chapter 11. Holographic Motility Contrast Imaging of Live Tissues

11.1 Introduction and Review

11.2 Optical Coherence Imaging

11.3 Motility Contrast Imaging

11.4 Dynamic Light Scattering (DLS) Spectroscopy

11.5 Tissue Dynamics Spectroscopy



Part 3: Advanced Interferometric and Polarization Techniques

Chapter 12. Tomographic Phase Microscopy (TPM)

12.1 Introduction

12.2 Theory—Optical Diffraction Tomography

12.3 Experimental Implementation

12.4 Video-Rate TPM

12.5 Biological Applications

12.6 Conclusion


Chapter 13. Phase-Sensitive Optical Coherence Microscopy (OCM)

13.1 Introduction

13.2 OCM Principles

13.3 Phase OCM

13.4 Phase OCM Applications

13.5 Conclusions


Chapter 14. Polarization and Spectral Interferometric Techniques for Quantitative Phase Microscopy

14.1 Introduction

14.2 Dual-Interference Channel Quantitative Phase Microscopy

14.3 Spectral-Domain Differential Interference Contrast Microscopy

14.4 Spectral Multiplexing by RGB Color Channels

14.5 Nonlinear Phase Dispersion Spectroscopy

14.6 Conclusions



Chapter 15. Polarization Microscopy

15.1 Introduction

15.2 Traditional Polarized Light Microscopy

15.3 The Liquid-Crystal Polarization Microscope (LC-PolScope)

15.4 Practical Considerations

15.5 Polarized Light Imaging in Three Dimensions

15.6 Conclusion

15.7 Glossary of Polarization Optical Terms



Part 4: Phase Nanoscopy

Chapter 16. Is There a Fundamental Limit to Spatial Resolution in Phase Microscopy?

16.1 Introduction

16.2 Where Was Abbe’s Theory Incomplete?

16.3 Parallel Full-Field Linear Imaging

16.4 Reconstruction from Diffraction Patterns

16.5 Structured Illumination Microscopy

16.6 Imaging Three-Dimensional Phase Objects

16.7 Conclusions



Chapter 17. Nano-Holographic Interferometry for In-Vivo Observations

17.1 Introduction: Theoretical Basis of Nano Fourier Transform Holography

17.2 Gratings Illuminated by Evanescent Waves

17.3 Basic Setup Utilized for the Observation of Nano-Objects

17.4 System of Fringes Contained in the Recorded Image: Multi-k Vector Fields

17.5 Determination of the Pitch of the Gratings

17.6 Formation of the Holograms at the Nanoscale

17.7 Procedures to Extract Metrological Information from the Recorded Images

17.8 Analysis of the Scales

17.9 Observation of Nano-Sized Objects

17.10 Conclusions


Chapter 18. Fluorescence Phase Microscopy (FPM) and Nanoscopy

18.1 Introduction

18.2 The Fluorescence Self-Interference Process

18.3 Experimental Setups of FPM

18.4 Applications of FPM

18.5 Conclusions and Outlook




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© Academic Press 2012
Academic Press
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About the Editor

Natan T. Shaked

Affiliations and Expertise

Senior Lecturer, Department of Biomedical Engineering, Tel Aviv University

Zeev Zalevsky

Affiliations and Expertise

Faculty of Engineering and the Center for Advanced Materials and Nanotechnology, Bar-Ilan University

Lisa Satterwhite

Affiliations and Expertise

Duke University, Durham


"This interesting and inspiring book is a collection of articles on specific applications in optical imaging of cells and tissues…Phase imaging is understood in a broad sense and related techniques of super-resolution, polarization and optical coherence microscopy are described in a concise but very informative manner."--Optics & Photonics News Online, June 13, 2013

"This book has been structured to be self-contained and therefore useful for researchers and students at all levels who need a cutting edge review of techniques and applications, particularly biological and medical applications."--Reference and Research Book News, December 2012