Superresolution Optical Microscopy

1. Front Matter

   Pages i-xxii

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2. Development of Microscopes

   1. Front Matter

      Pages 1-3

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   2. Connections Between Light, Vision, and Microscopes

      • Barry R. Masters

      Pages 5-12

   3. Concepts and Criteria of Resolution

      • Barry R. Masters

      Pages 13-32

   4. Aberrations and Artifacts Confound Optical Resolution

      • Barry R. Masters

      Pages 33-39

   5. Insights into the Development of Light Microscopes

      • Barry R. Masters

      Pages 41-50

   6. Ernst Abbe and His Contributions to Optics

      • Barry R. Masters

      Pages 51-64

   7. Abbe’s Theory of Image Formation in the Microscope

      • Barry R. Masters

      Pages 65-108

   8. Helmholtz’s Contributions on the Theoretical Limits to the Resolution of the Microscope

      • Barry R. Masters

      Pages 109-139

   9. Further Insights into Abbe’s Theory of Image Formation in the Microscope Based on Diffraction

      • Barry R. Masters

      Pages 141-151

   10. Mathematical Description of Abbe’s Theory of Image Formation in the Microscope Based on Diffraction

       • Barry R. Masters

       Pages 153-161

3. Optical Techniques to Enhance Contrast in the Microscope

   1. Front Matter

      Pages 163-164

      PDF

   2. Richard Zsigmondy and Henry Siedentopf’s Ultramicroscope

      • Barry R. Masters

      Pages 165-172

   3. Light-Sheet Fluorescence Microscopy

      • Barry R. Masters

      Pages 173-211

   4. Phase Microscopy to Enhance Contrast

      • Barry R. Masters

      Pages 213-228

4. Far-Field Superresolution Optical Microscopy

   1. Front Matter

      Pages 229-232

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   2. Structured Illumination Microscopy

      • Barry R. Masters

      Pages 233-260

   3. Stimulated Emission Depletion Microscopy and Related Techniques

      • Barry R. Masters

      Pages 261-305

   4. Localization Microscopy with Active Control

      • Barry R. Masters

      Pages 307-369

   5. Coda: Trade-Offs, Cautions, and Limitations of Superresolution Optical Microscopes

      • Barry R. Masters

      Pages 371-377

This book presents a comprehensive and coherent summary of techniques for enhancing the resolution and image contrast provided by far-field optical microscopes. It takes a critical look at the body of knowledge that comprises optical microscopy, compares and contrasts the various instruments, provides a clear discussion of the physical principles that underpin these techniques, and describes advances in science and medicine for which superresolution microscopes are required and are making major contributions. 

The text fills significant gaps that exist in other works on superresolution imaging, firstly by placing a new emphasis on the specimen, a critical component of the microscope setup, giving equal importance to the enhancement of both resolution and contrast. Secondly, it covers several topics not typically discussed in depth, such as Bessel and Airy beams, the physics of the spiral phase plate, vortex beams and singular optics, photoactivated localizationmicroscopy (PALM), stochastic optical reconstruction microscopy (STORM), structured illumination microscopy (SIM), and light-sheet fluorescence microscopy (LSFM). Several variants of these techniques are critically discussed. Noise, optical aberrations, specimen damage, and artifacts in microscopy are also covered. The importance of validation of superresolution images with electron microscopy is stressed. Additionally, the book includes translations and discussion of seminal papers by Abbe and Helmholtz that proved to be pedagogically relevant as well as historically significant. 


This book is written for students, researchers, and engineers in the life sciences, medicine, biological engineering, and materials science who plan to work with or already are working with superresolution light microscopes. The volume can serve as a reference for these areas while a selected set of individual chapters can be used as a textbook for a one-semester undergraduate or first-year graduate course on superresolution microscopy. Moreover, the text provides a captivating account of curiosity, skepticism, risk-taking, innovation, and creativity in science and technology. Good scientific practice is emphasized throughout, and the author’s lecture slides on responsible conduct of research are included as an online resource which will be of interest to students, course instructors, and scientists alike.

• superresolution microscopy
• light-sheet microscopy
• nondiffractive beams
• structured illumination microscopy
• angular momentum of light
• nonlinear light microscopy
• resolution in microscopy
• contrast in microscopy
• aberrations in microscopy
• molecular spectroscopy
• Biological Microscopy

“The book is a must for anyone working in optics as well as for anyone interested in science in general. It is very clearly written, touches on many historical aspects of optics and has many photos supporting the text information.” (Daniela Dragoman, Optics & Photonics News, osa-opn.org, July 16, 2020)

• Previously, Visiting Scientist Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, USA

  Barry R. Masters

Professor Barry R. Masters received a PhD from The Weizmann Institute of Science, Israel, an MS degree from Florida State University, and a BS from The Polytechnic Institute of Brooklyn, New York. Professor Masters was Visiting Scientist in the Department of Biological Engineering at the Massachusetts Institute of Technology, Visiting Scholar in the History of Science Department at Harvard University, Visiting Professor in the Department of Ophthalmology at the University of Bern, and Professor in Anatomy and Cell Biology at the Uniformed Services University of the Health Sciences. He was a Visiting Professor in Japan, India, Australia, Switzerland, Taiwan, Turkey, The United Kingdom, P. R. China, and Egypt. He is a Fellow of the American Association for the Advancement of Science (AAAS), the Optical Society of America (OSA), and the international society for optics and photonics (SPIE). From 1999 to 2000, Professor Masters was an AAAS Congressional Science and Engineering Fellow and served as Legislative Assistant in the United States Congress. He is a recipient of the Vogt Prize for Ophthalmic Research (highest Swiss award for research in ophthalmology) with Dr. Matthias Böhnke in 1999 for their work on “confocal microscopy of the cornea.” Professor Masters has published 87 refereed papers, 143 book chapters and proceedings articles, 105 abstracts, and 239 book reviews. He is the editor or author of 10 books, among them Noninvasive Diagnostic Techniques in Ophthalmology, Confocal Microscopy and Multiphoton Excitation Microscopy: The Genesis of Live Cell Imaging, and Handbook of Biomedical Nonlinear Optical Microscopy (with Peter So). Masters wrote the essay “What Is Light?” (translated into 15 languages) for the International Commission for Optics (ICO), to inaugurate the International Year of Light, 2015. He lectures worldwide on the Responsible Conduct of Research, biomedical ethics, critical thinking, and biomedical photonics. His scholarly interests include developments in the fields of in vivo microscopy of the human eye and skin, biomedical imaging and spectroscopy, fractal analysis of branching vascular patterns, ethics in science and medicine, the history of nineteenth- and twentieth-century physics, science education, and art and science.

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