Atomic Force Microscopy For Biologists

Atomic force microscopy (AFM) is part of a range of emerging microscopic methods for biologists which offer the magnification range of both the light and electron microscope, but allow imaging under the "natural" conditions usually associated with the light microscope. To biologists, AFM offers the prospect of high resolution images of biological material, images of molecules and their interactions even under physiological conditions, and the study of molecular processes in living systems. This book provides a realistic appreciation of the advantages and limitations of the technique and the present and future potential for improving the understanding of biological systems. The second edition of this bestseller has been updated to describe the latest developments in this exciting field, including a brand new chapter on force spectroscopy. The dramatic developments of AFM over the past ten years from a simple imaging tool to the multi-faceted, nano-manipulating technique that it is today are conveyed in a lively and informative narrative, which provides essential reading for students and experienced researchers alike. Contents:ApparatusBasic PrinciplesMacromoleculesInterfacial SystemsOrdered MacromoleculesCells, Tissue and BiomineralsOther Probe MicroscopesForce Spectroscopy Readership: Undergraduates, postgraduates and researchers in biophysics. Keywords:AFM;Scanning Probe Microscopy (SPM);Biophysics;Cells;Macromolecules;Imaging;Biopolymers;Interfaces;DNA;PhospholipidsKey Features:Over 70 new pages, 700 new references, bringing it bang up-to-date with current literature, and 24 new figuresOnly book covering published research, machines and methodology from inception to the present day (2009)Updated to include and an entirely new chapter on force spectroscopyReviews:“This second edition of an excellent book updates considerably the information contained, and is expanded, too. The main focus of this book are the biological applications of AFM, and these are covered very well … The chapters are in-depth and very informative, and contain lots of useful and detailed information. In general the book is well written with an informal style, and contains useful information for beginners, including detailed information on how to carry out some experiments, answers to common questions, etc. … Overall this book is highly recommended for those wishing to get an overview of the biological applications of AFM.”Peter Eaton University of Porto, Portugal Reviews of the First Edition “This book fills an important niche by providing an introduction to AFM that will be understandable to a wide range of scientists … the reader is offered an introduction of sufficient brevity to actually be read and of sufficient depth to help in defining the potential uses of SPMs in his or her own research.” Scott Fraser (California Institute of Technology, USA) Nature Cell Biology “I found this book easy to read with clear, simple explanations of a complex topic. I think that the selection of the subjects treated in the book is sufficient for the non-specialist to understand both the basic concepts of the AFM and the production of AFM images in biological systems.”Microscopy and Analysis “… is an excellent introduction for anybody wishing to enter this field … In summary I would strongly recommend this book to any biologist planning on carrying out research using SPM imaging techniques and existing users in the field who wish to broaden their knowledge of SPM imaging and the research already carried out on common biological systems.”Drew Murray JPK Instruments, Berlin “This book provides an excellent survey of novel applications. It is nicely illustrated with numerous images from leading experts in the field. Clear descriptions of the apparatus and its basic principles are provided in a way accessible to students/scientists who do not have a strong physics background. It will be useful to biologists, but also non-biologists dealing with biosystems …”Prof Y F Dufrêne UCL, Belgium

Ch. 1. An introduction -- ch. 2. Apparatus. 2.1. The atomic force microscope. 2.2. Piezoelectric scanners. 2.3. Probes and cantilevers. 2.4. Sample holders. 2.5. Detection methods. 2.6. Control systems. 2.7. Vibration isolation : thermal and mechanical. 2.8. Calibration. 2.9. Integrated AFMs -- ch. 3. Basic principles. 3.1. Forces. 3.2. Imaging modes. 3.3. Image types. 3.4. Substrates. 3.5. Common problems. 3.6. Getting started. 3.7. Image optimisation -- ch. 4. Macromolecules. 4.1. Imaging methods. 4.2. Nucleic acids : DNA. 4.3. Nucleic acids : RNA. 4.4. Polysaccharides. 4.5. Proteins -- ch. 5. Interfacial systems. 5.1. Introduction to interfaces. 5.2. Sample preparation. 5.3. Phospholipids. 5.4. Liposomes and intact vesicles. 5.5. Lipid-protein mixed films. 5.6. Miscellaneous lipid films/surfactant films. 5.7. Interfacial protein films -- ch. 6. Ordered macromolecules. 6.1. Interfacial protein films. 6.2. Two dimensional protein crystals : an introduction. 6.3. AFM studies of 2D membrane protein crystals. 6.4. AFM studies of 2D crystals of soluble proteins -- ch. 7. Cells, tissue and biominerals. 7.1. Imaging methods. 7.2. Microbial cells : bacteria, spores and yeasts. 7.3. Blood cells. 7.4. Neurons and Glial cells. 7.5. Epithelial cells. 7.6. Non-confluent renal cells. 7.7. Endothelial cells. 7.8. Cardiocytes. 7.9. Other mammalian cells. 7.10. Plant cells. 7.11. Tissue. 7.12. Biominerals -- ch. 8. Other probe microscopes. 8.1. Overview. 8.2. Scanning tunnelling microscope (STM). 8.3. Scanning near-field optical microscope (SNOM). 8.4. Scanning ion conductance microscope (SICM). 8.5. Scanning thermal microscope (SThM). 8.6. Optical tweezers and the photonic force microscope (PPM) -- ch. 9. Force spectroscopy. 9.1. Force measurement with the AFM. 9.2. First steps in force spectroscopy: from raw data to force-distance curves. 9.3. Pulling methods. 9.4. Pushing methods. 9.5. Analysis of force-distance curves

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 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.

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 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).

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.