Volume 55 in Methods in Cell Biology is a concise laboratory book that emphasizes the methods and technologies needed to use single polarized laser light source that functions simultaneously as an optical trap and a dual-beam interferometer.* * Provides a practical laboratory guide for methods and technologies used with laser tweezers* Includes comprehensive and easy-to-follow protocols
Volume 55 in Methods in Cell Biology is a concise laboratory book that emphasizes the methods and technologies needed to use single polarized laser light source that functions simultaneously as an optical trap and a dual-beam interferometer. * * Provides a practical laboratory guide for methods and technologies used with laser tweezers * Includes comprehensive and easy-to-follow protocols
A comprehensive guide to the theory, practice and applications of optical tweezers, combining state-of-the-art research with a strong pedagogic approach.
There are probably few people who do not dream of the good old times, when do ing science often meant fascination, excitement, even adventure. In our time, do ing science involves often technology and, perhaps, even business. But there are still niches where curiosity and fascination have their place. The subject of this book, technological as its title may sound, is one of the fortunate examples. It will report on lasers generating the coldest places in the Universe, and on table top laser microtools which can produce a heat "inferno" as it prevails in the interior of the Sun, or simulate, for specific plant cells, microgravity of the space around our plan et Earth. There will be some real surprises for the reader. The applications range from basic studies of the driving forces of cell division (and thus life) via genetic modification of cells (for example, for plant breeding) to medical applications such as blood cell analysis and finally in vitro fertilization. What are these instruments: laser microbeams and optical tweezers? Both are lasers coupled with a fluorescence microscope. The laser microbeam uses a pulsed ultraviolet laser. Light is focused, as well as possible, in space and time, in order to obtain extremely high light intensities - high enough to generate, for a very short instant, extremely hot spots which can be used to cut, fuse or perforate biological material.
This important volume contains selected papers and extensive commentaries on laser trapping and manipulation of neutral particles using radiation pressure forces. Such techniques apply to a variety of small particles, such as atoms, molecules, macroscopic dielectric particles, living cells, and organelles within cells. These optical methods have had a revolutionary impact on the fields of atomic and molecular physics, biophysics, and many aspects of nanotechnology.In atomic physics, the trapping and cooling of atoms down to nanokelvins and even picokelvin temperatures are possible. These are the lowest temperatures in the universe. This made possible the first demonstration of Bose-Einstein condensation of atomic and molecular vapors. Some of the applications are high precision atomic clocks, gyroscopes, the measurement of gravity, cryptology, atomic computers, cavity quantum electrodynamics and coherent atom lasers.A major application in biophysics is the study of the mechanical properties of the many types of motor molecules, mechanoenzymes, and other macromolecules responsible for the motion of organelles within cells and the locomotion of entire cells. Unique in vitro and in vivo assays study the driving forces, stepping motion, kinetics, and efficiency of these motors as they move along the cell's cytoskeleton. Positional and temporal resolutions have been achieved, making possible the study of RNA and DNA polymerases, as they undergo their various copying, backtracking, and error correcting functions on a single base pair basis.Many applications in nanotechnology involve particle and cell sorting, particle rotation, microfabrication of simple machines, microfluidics, and other micrometer devices. The number of applications continues to grow at a rapid rate.The author is the discoverer of optical trapping and optical tweezers. With his colleagues, he first demonstrated optical levitation, the trapping of atoms, and tweezer trapping and manipulation of living cells and biological particles.This is the only review volume covering the many fields of optical trapping and manipulation. The intention is to provide a selective guide to the literature and to teach how optical traps really work.
The technical development of optical tweezers, along with their application in the biological and physical sciences, has progressed significantly since the demonstration of an optical trap for micron-sized particles based on a single, tightly focused laser beam was first reported more than twenty years ago. Bringing together many landmark papers on
This monograph focuses on modern femtosecond laser microscopes for two photon imaging and nanoprocessing, on laser tweezers for cell micromanipulation as well as on fluorescence lifetime imaging (FLIM) in Life Sciences. The book starts with an introduction by Dr. Wolfgang Kaiser, pioneer of nonlinear optics and ends with the chapter on clinical multiphoton tomography, the novel high resolution imaging technique. It includes a foreword by the nonlinear microscopy expert Dr. Colin Sheppard. Contents Part I: Basics Brief history of fluorescence lifetime imaging The long journey to the laser and its use for nonlinear optics Advanced TCSPC-FLIM techniques Ultrafast lasers in biophotonics Part II: Modern nonlinear microscopy of live cells STED microscopy: exploring fluorescence lifetime gradients for super-resolution at reduced illumination intensities Principles and applications of temporal-focusing wide-field two-photon microscopy FLIM-FRET microscopy TCSPC FLIM and PLIM for metabolic imaging and oxygen sensing Laser tweezers are sources of two-photon effects Metabolic shifts in cell proliferation and differentiation Femtosecond laser nanoprocessing Cryomultiphoton imaging Part III: Nonlinear tissue imaging Multiphoton Tomography (MPT) Clinical multimodal CARS imaging In vivo multiphoton microscopy of human skin Two-photon microscopy and fluorescence lifetime imaging of the cornea Multiscale correlative imaging of the brain Revealing interaction of dyes and nanomaterials by multiphoton imaging Multiphoton FLIM in cosmetic clinical research Multiphoton microscopy and fluorescence lifetime imaging for resection guidance in malignant glioma surgery Non-invasive single-photon and multi-photon imaging of stem cells and cancer cells in mouse models Bedside assessment of multiphoton tomography
Physical Biology of the Cell is a textbook for a first course in physical biology or biophysics for undergraduate or graduate students. It maps the huge and complex landscape of cell and molecular biology from the distinct perspective of physical biology. As a key organizing principle, the proximity of topics is based on the physical concepts that
Advances in technology have revolutionized the development of light microscopy techniques in biomedical research, thus improving visualization of the microstructure of cells and tissues under physiological conditions. Fluorescence microscopy methods are non-contact and non-invasive and provide high spatial and temporal resolution that other laboratory techniques cannot. This well-illustrated book targets graduate students and scientists who are new to the state-of-the-art fluorescence microscopy techniques used in biological and clinical imaging. It explains basic concepts and imaging procedures for wide-field, confocal, multiphoton excitation, fluorescence resonance energy transfer (FRET), lifetime imaging (FLIM), spectral imaging, fluorescence recovery after photobleaching (FRAP), optical tweezers, total internal reflection, high spatial resolution atomic force microscopy (AFM), and bioluminescence imaging for gene expression. The usage of these techniques in various biological applications, including calcium, pH, membrane potential, mitochondrial signaling, protein-protein interactions under various physiological conditions, and deep tissue imaging, is clearly presented. The authors describe the approaches to selecting epifluorescence microscopy, the detectors, and the image acquisition and processing software for different biological applications. Step-by-step directions on preparing different digital formats for light microscopy images on websites are also provided.
This volume is essential for anyone struggling with the cellular heterogeneity field, and it connects this new area with the latest advances in innovative techniques combined with practical advice from international experts. The chapters provide a comprehensive overview of cellular heterogeneity analysis techniques, exploring cytometry, imaging and spectroscopy methods, metabolic and molecular biological methods in tracking, as well as chromatin and cell cycle heterogeneity. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, detailed lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Cellular Heterogeneity: Methods and Protocols encourages readers to explore new ways of studying cells that will help lead to exciting new discoveries.
