Atomic Force Microscopy In Cell Biology

This is the first book to cover the history, structure, and application of atomic force microscopy in cell biology. Presented in the clear, well-illustrated style of the Methods in Cell Biology series, it introduces the AFM to its readers and enables them to tap the power and scope of this technology to further their own research. A practical laboratory guide for use of the atomic force and photonic force microscopes, it provides updated technology and methods in force spectroscopy. It is also a comprehensive and easy-to-follow practical laboratory guide for the use of the AFM and PFM in biological research.

The book addresses new achievements in AFM instruments – e.g. higher speed and higher resolution – and how AFM is being combined with other new methods like NSOM, STED, STORM, PALM, and Raman. This book explores the latest advances in atomic force microscopy and related techniques in molecular and cell biology. Atomic force microscopy (AFM) can be used to detect the superstructures of the cell membrane, cell morphology, cell skeletons and their mechanical properties. Opening up new fields of in-situ dynamic study for living cells, enzymatic reactions, fibril growth and biomedical research, these combined techniques will yield valuable new insights into molecule and cell biology. This book offers a valuable resource for students and researchers in the fields of biochemistry, cell research and chemistry etc.

The atomic force microscope (AFM) has become one of the leading nanoscale measurement techniques for materials science since its creation in the 1980's, but has been gaining popularity in a seemingly unrelated field of science: biology. The AFM naturally lends itself to investigating the topological surfaces of biological objects, from whole cells to protein particulates, and can also be used to determine physical properties such as Young's modulus, stiffness, molecular bond strength, surface friction, and many more. One of the most important reasons for the rise of biological AFM is that you can measure materials within a physiologically relevant environment (i.e. liquids). This book is a collection of works beginning with an introduction to the AFM along with techniques and methods of sample preparation. Then the book displays current research covering subjects ranging from nano-particulates, proteins, DNA, viruses, cellular structures, and the characterization of living cells.

This first book on high-speed atomic force microscopy (HS-AFM) is intended for students and biologists who want to use HS-AFM in their research. It provides straightforward explanations of the principle and techniques of AFM and HS-AFM. Numerous examples of HS-AFM studies on proteins demonstrate how to apply this new form of microscopy to specific biological problems. Several precautions for successful imaging and the preparation of cantilever tips and substrate surfaces will greatly benefit first-time users of HS-AFM. In turn, the instrumentation techniques detailed in Chapter 4 can be skipped, but will be useful for engineers and scientists who want to develop the next generation of high-speed scanning probe microscopes for biology. The book is intended to facilitate the first-time use of this new technique, and to inspire students and researchers to tackle their own specific biological problems by directly observing dynamic events occurring in the nanoscopic world. Microscopy in biology has recently entered a new era with the advent of high-speed atomic force microscopy (HS-AFM). Unlike optical microscopy, electron microscopy, and conventional slow AFM, it allows us to directly observe biological molecules in physiological environments. Molecular “movies” created using HS-AFM can directly reveal how molecules behave and operate, without the need for subsequent complex analyses and roundabout interpretations. It also allows us to directly monitor morphological change in live cells, and dynamic molecular events occurring on the surfaces of living bacteria and intracellular organelles. As HS-AFM instruments were recently commercialized, in the near future HS-AFM is expected to become a common tool in biology, and will enhance and accelerate our understanding of biological phenomena.

Recent developments in atomic force microscopy (AFM) have been accomplished through various technical and instrumental innovations, including high-resolution and recognition imaging technology under physiological conditions, fast-scanning AFM, and general methods for cantilever modification and force measurement. All these techniques are now highly

A complete examination of the uses of the atomic force microscope in biology and medicine This cutting-edge text, written by a team of leading experts, is the first detailed examination of the latest, most powerful scanning probe microscope, the atomic force microscope (AFM). Using the AFM, in combination with conventional tools and techniques, readers gain a profound understanding of the cell, subcellular organelles, and biomolecular structure and function. The text begins with three chapters describing the molecular machinery and mechanism of cell secretion and membrane fusion in cells, using approaches that combine AFM, electron microscopy, X-ray diffraction, photon correlation spectroscopy, molecular biology, biochemistry, and electrophysiology. The discovery of a new cellular structure the "porosome" or fusion pore--the cells secretory machinery, the molecular mechanism of membrane fusion in cells, and the expulsion of intravesicular contents during cell secretion are outlined in the first three chapters. The book also covers: * Identification of the "porosome" in the growth hormone secreting cell of the pituitary gland * Probing the structural and physical properties of microbial cell surfaces * Scanning probe microscopic characterization of the higher plant cell wall and its components * Case studies of nano drug delivery systems using engineered dendrimers * AFM techniques for studying living cells * Investigating the intermolecular forces of leukocyte adhesion molecules * Protein-protein interactions * Micromechanical properties of lipid bilayers and vesicles The text concludes with four chapters that examine new and emerging approaches in the use of force microscopy in biology and medicine. This text is ideal for advanced undergraduate and graduate students and researchers in cell and molecular biology, genetics, genomics, physiology, neuroscience, biophysics, and biochemistry. Not only does it provide the theory, but also practical considerations such as the selection of the right tools and approach.

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).

Despite substantial evidence showing the feasibility of Atomic Force Microscopy (AFM) to identify cells with altered elastic and adhesive properties, the use of this technique as a complementary diagnostic method remains controversial. This book is designed to be a practical textbook that teaches how to assess the mechanical characteristics of living, individual cells by AFM. Following a step-by-step approach, it introduces the methodology of measurements in the case of both determination of elastic properties and quantification of adhesive properties.