Biomedical Light Microscopy

New interest in light microscopy of the last few years has not been backed up by adequate general literature. This book intends to fill the gap between specialized texts on detailed topics and general introductory booklets, mostly dealing with the use of the conventional light microscope only. In this short textbook both new developments in microscopy and basic facts of image formation will be treated, including often neglected topics such as axial resolving power, lens construction, photomicrography and correct use of phase-en interference contrast systems. Theoretical background will be dealt with as far as necessary for a well-considered application of these techniques enabling a deliberate choice for the approach of a certain problem. Over 150 illustrations (photomicrographs and diagrams) complete the information on microscopy of the nineties in the biomedical field, intended for scientists, doctors, technicians and research students. Many drawings have been contributed by the illustrator R. Kreuger; the photographic work has been executed by J. Peeterse. Secretarial assistance in preparing the manuscript was given by Ms T. M. S. Pierik. Dr M. J. Pearson has corrected the English of the final text.

The second volume of the series Manuals in Biomedical Research, this book is aimed to be both a concise introduction to the diverse field of microscopy and a practical guide those who require the use of microscopic for methods in their research. It provides young as well as experienced scientists a state-of-the-art multidisciplinary overview of microscopic techniques, covering all the major microscopy fields in biomedical sciences and showing their application in evaluating samples ranging from molecules to cells and tissues.Microscopy has revolutionized our understanding of biological events. Within the last two decades, microscopic techniques have provided insights into the dynamics of biological processes that regulate such events. Biological discovery, to a large extent, depends on advances in imaging techniques and various microscopic techniques have emerged as central and indispensable tools in the biomedical sciences.The four authors bring with them extensive experiences spanning across disciplines such as Microbiology, Molecular and Cell Biology, Tissue Engineering, Biomedical and Regenerative Medicine and so forth, reinforcing the fact that microscopy has proven useful in countless investigations into the mysteries of life.

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. 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.

The Handbook of Biomedical Nonlinear Optical Microscopy provides comprehensive treatment of the theories, techniques, and biomedical applications of nonlinear optics and microscopy for cell biologists, life scientists, biomedical engineers, and clinicians. The chapters are separated into basic and advanced sections, and provide both textual and graphical illustrations of all key concepts. The more basic sections are aimed at life scientists without advanced training in physics and mathematics, and tutorials are provided for the more challenging sections. The first part of the Handbook introduces the historical context of nonlinear microscopy. The second part presents the nonlinear optical theory of two- and multiphoton excited fluorescence (TPE, MPE) spectroscopy, second and third harmonic generation (SHG, THG) spectroscopy, and coherent anti-Stokes Raman spectroscopy (CARS). The third part introduces modern microscopic and spectroscopic instrumentation and techniques that are based on nonlinear optics. The fourth part provides key applications of nonlinear microscopy to the biomedical area: neurobiology, immunology, tumor biology, developmental biology, dermatology, and cellular metabolism. There are also chapters on nonlinear molecular probes, cellular damage, and nanoprocessing.

Ideal for cell biologists, life scientists, biomedical engineers, and clinicians, this handbook provides comprehensive treatment of the theories, techniques, and biomedical applications of nonlinear optics and microscopy.

This open access book provides a comprehensive overview of the application of the newest laser and microscope/ophthalmoscope technology in the field of high resolution imaging in microscopy and ophthalmology. Starting by describing High-Resolution 3D Light Microscopy with STED and RESOLFT, the book goes on to cover retinal and anterior segment imaging and image-guided treatment and also discusses the development of adaptive optics in vision science and ophthalmology. Using an interdisciplinary approach, the reader will learn about the latest developments and most up to date technology in the field and how these translate to a medical setting. High Resolution Imaging in Microscopy and Ophthalmology – New Frontiers in Biomedical Optics has been written by leading experts in the field and offers insights on engineering, biology, and medicine, thus being a valuable addition for scientists, engineers, and clinicians with technical and medical interest who would like to understand the equipment, the applications and the medical/biological background. Lastly, this book is dedicated to the memory of Dr. Gerhard Zinser, co-founder of Heidelberg Engineering GmbH, a scientist, a husband, a brother, a colleague, and a friend.

Biomedical optical imaging is a rapidly emerging research area with widespread fundamental research and clinical applications. This book gives an overview of biomedical optical imaging with contributions from leading international research groups who have pioneered many of these techniques and applications. A unique research field spanning the microscopic to the macroscopic, biomedical optical imaging allows both structural and functional imaging. Techniques such as confocal and multiphoton microscopy provide cellular level resolution imaging in biological systems. The integration of this technology with exogenous chromophores can selectively enhance contrast for molecular targets as well as supply functional information on processes such as nerve transduction. Novel techniques integrate microscopy with state-of-the-art optics technology, and these include spectral imaging, two photon fluorescence correlation, nonlinear nanoscopy; optical coherence tomography techniques allow functional, dynamic, nanoscale, and cross-sectional visualization. Moving to the macroscopic scale, spectroscopic assessment and imaging methods such as fluorescence and light scattering can provide diagnostics of tissue pathology including neoplastic changes. Techniques using light diffusion and photon migration are a means to explore processes which occur deep inside biological tissues and organs. The integration of these techniques with exogenous probes enables molecular specific sensitivity.

The natural, biological, medical, and related sciences would not be what they are today without the microscope. After the introduction of the optical microscope, a second breakthrough in morphostructural surface analysis occurred in the 1940s with the development of the scanning electron microscope (SEM), which, instead of light (i. e. , photons) and glass lenses, uses electrons and electromagnetic lenses (magnetic coils). Optical and scanning (or transmission) electron microscopes are called “far-field microscopes” because of the long distance between the sample and the point at which the image is obtained in comparison with the wavelengths of the photons or electrons involved. In this case, the image is a diffraction pattern and its resolution is wavelength limited. In 1986, a completely new type of microscopy was proposed, which, without the use of lenses, photons, or electrons, directly explores the sample surface by means of mechanical scanning, thus opening up unexpected possibilities for the morphostructural and mechanical analysis of biological specimens. These new scanning probe microscopes are based on the concept of near-field microscopy, which overcomes the problem of the limited diffraction-related resolution inherent in conventional microscopes. Located in the immediate vicinity of the sample itself (usually within a few nanometers), the probe records the intensity, rather than the interference signal, thus significantly improving resolution. Since the most we- known microscopes of this type operate using atomic forces, they are frequently referred to as atomic force microscopes (AFMs).