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.
Proceeding from basic fundamentals to applications, this volume provides a comprehensive overview of the use of AFM and related scanning probe microscopies for cell surface analysis. It covers all cell types, from viruses and protoplasts to bacteria and animal cells. It also discusses a range of advanced AFM modalities, including high-resolution imaging, nanoindentation measurements, recognition imaging, and single-molecule and single-cell force spectroscopy. The book covers methodologies for preparing and analyzing cells and membranes of all kinds and highlights recent examples to illustrate the power of AFM techniques in life sciences and nanomedicine.
This is the first book to bring together both the basic theory and proven process engineering practice of AFM. It is presented in a way that is accessible and valuable to practising engineers as well as to those who are improving their AFM skills and knowledge, and to researchers who are developing new products and solutions using AFM. The book takes a rigorous and practical approach that ensures it is directly applicable to process engineering problems. Fundamentals and techniques are concisely described, while specific benefits for process engineering are clearly defined and illustrated. Key content includes: particle-particle, and particle-bubble interactions; characterization of membrane surfaces; the development of fouling resistant membranes; nanoscale pharmaceutical analysis; nanoengineering for cellular sensing; polymers on surfaces; micro and nanoscale rheometry. - Atomic force microscopy (AFM) is an important tool for process engineers and scientists as it enables improved processes and products - The only book dealing with the theory and practical applications of atomic force microscopy in process engineering - Provides best-practice guidance and experience on using AFM for process and product improvement
This book presents methodological and application research in detecting cellular and molecular biophysical properties based on atomic force microscopy (AFM) nanorobotics. Series methods for in situ label-free visualizing and quantifying the multiple physical properties of single cells and single molecules were developed, including immobilization strategies for observing fine structures of living cells, measurements of single-cell mechanics, force recognition of molecular interactions, and mapping protein organizations on cell surface. The biomedical applications of these methods in clinical lymphoma treatments were explored in detail, including primary sample preparation, cancer cell recognition, AFM detection and data analysis. Future directions about the biomedical applications of AFM are also given.
Over the last two decades, Atomic Force Microscopy (AFM) has undoubtedly had a considerable impact in unraveling the structures and dynamics of microbial surfaces with nanometer resolution, and under physiological conditions. Moreover, the continuous innovations in AFM-based modalities as well as the combination of AFM with modern optical techniques, such as confocal fluorescence microscopy or Raman spectroscopy, increased the diversity and volume of data that can be acquired in an experiment. It is evident that these combinations provide new ways to investigate a broad spectrum of microbiological processes at the level of single cells. In this book, I have endeavored to highlight the wealth of AFM-based modalities that have been implemented over the recent years leading to the multiparametric and multifunctional characterization of, specifically, bacterial surfaces. Examples include the real-time imaging of the nanoscale organization of cell walls, the quantification of subcellular chemical heterogeneities, the mapping and functional analysis of individual cell wall constituents, and the probing of the nanomechanical properties of living bacteria. It is expected that in the near future more AFM-based modalities and complementary techniques will be combined into single experiments to address pertinent problems and challenges in microbial research. Such improvements may make it possible to address the dynamic nature of many more microbial cell surfaces and their constituents, including the restructuring of cellular membranes, pores and transporters, signaling of cell membrane receptors, and formation of cell-adhesion complexes. Ultimately, manifold discoveries and engineering possibilities will materialize as multiparametric tools allow systems of increasing complexity to be probed and manipulated.
With its ability to explore the surface of the sample by means of a local scanning probe and its use of dedicated software allows to be visualize results, atomic force microscopy (AFM) has revolutionized the study of the smallest aspects of life. Atomic Force Microscopy in Biomedical Research: Methods and Protocols proves that this technology is no longer simply just another form of microscopy but has given rise to a completely new way of using microscopy that fulfils the dreams of all microscopists: being able to touch, move, and interact with the sample while it is being examined, thus making it possible to discover not only morphological but also chemical and physical structural information. Covering such topics as molecule imaging, nanoscale surface analysis and cellular imaging, force-spectroscopy, investigating drug action, and AFM as a nanotool, this volume features the most up-to-date techniques currently in use. Written in the Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials, step-by-step, readily reproducible protocols, and expert tips on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, Atomic Force Microscopy in Biomedical Research: Methods and Protocols brings together different types of applications in order to provide examples from diverse fields in the hope of inspiring researchers to apply their ingenuity in their own specialization and add significant originality to their varying studies.
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).
Summarizing the latest trends and the current state of this research field, this up-to-date book discusses in detail techniques to perform localized alterations on surfaces with great flexibility, including microfluidic probes, multifunctional nanopipettes and various surface patterning techniques, such as dip pen nanolithography. These techniques are also put in perspective in terms of applications and how they can be transformative of numerous (bio)chemical processes involving surfaces. The editors are from IBM Zurich, the pioneers and pacesetters in the field at the forefront of research in this new and rapidly expanding area.
