The mechanical response of a cell to external forces carries information about its structure and function. Because cell manipulation should ideally be non-invasive while performing sophisticated biophysical characterization, the radiation force of optical tweezers has become highly attractive. In this thesis, we explore three types of recently-developed optical tweezers: 1) static, 2) time-sharing and 3) oscillating. Using a full three-dimensional finite element method (3DFEM), modeling of each of these regimes allows us to fit experiments and access the cell mechanical properties. Combining optical trapping with cell mechanics requires interdisciplinary efforts. A survey of the various experimental approaches for optical trapping and measurements on isolated cells is presented. We then lay the theoretical background linking the interaction of optical fields to the cell's mechanical response. We are the first to implement a 3DFEM calculation including light scattering and the radiation stress distribution to predict the deformation of a biconcave cell -emulating a red blood cell- in static dual-trap optical tweezers. At equilibrium, the final deformation is given by the separation distance of the two trapping beams, revealing how the cell can be elongated or shrunk. Time-sharing optical tweezers realize multiple traps to manipulate objects ranging from macromolecules to biological cells. Our quantitative analysis shows how, although jumping, the local stress and strain is omnipresent in the cell. The viscoelastic object deformation and internal energy dissipation are analyzed. Another cell shape, a cubic rod, is also studied, elucidating novel symmetrical properties of the mechanical response. Finally, the analysis of the time-dependent deformation -creep testing- of a cell in static and time-sharing optical tweezers, shows that deformation of the object depends altogether on the object's viscoelasticity, significantly on its 3D shape and the mechanical loading. However, dynamic testing with oscillating optical tweezers surprisingly shows a phase shift between the loading stress (external force) and strain (deformation) independent on the 3D cell shape. This is a novel avenue giving access to the cell's viscoelasticity dynamic complex modulus directly in the time-domain.

A comprehensive guide to the theory, practice and applications of optical tweezers, combining state-of-the-art research with a strong pedagogic approach.

This detailed volume explores a wide variety of techniques involving optical tweezers, a technology that has become increasingly more accessible to a broad range of researchers. Beginning with recent technical advances, the book continues by covering the application of optical tweezers to study DNA-protein interactions and DNA motors, protocols to perform protein (un)folding experiments, the application of optical tweezers to study actin- and microtubule-associated motor proteins, and well as protocols for investigating the function and mechanical properties of microtubules and intermediate filaments, and more. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Optical Tweezers: Methods and Protocols, Second Edition serves as an ideal resource for expanding the accessibility and use of optical traps by scientists of diverse disciplines.

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.

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

Thanks to the pioneering works of Ashkin and coworkers, optical tweezers (OTs) have become an invaluable tool for myriad studies throughout the natural sciences. Their success relies on the fact that they can be considered as exceptionally sensitive transducers that are able to resolve pN forces and nm displacements, with high temporal resolution, down to μs. Hence their application to study a wide range of biological phenomena such as measuring the compliance of bacterial tails, the forces exerted by a single motor protein, and the mechanical properties of human red blood cells and of individual biological molecules. The number of articles related to them totals to a whopping 58,000 (source Google Scholar)! Microrheology is a branch of rheology, but it works at micrometer length scales and with microliter sample volumes. Therefore, microrheology techniques have been revealed to be very useful tools for all those rheological/mechanical studies where rare or precious materials are employed, such as in biological and biomedical studies. The aim of this book is to provide a pedagogical introduction to the physics principles governing both the optical tweezers and their application in the field of microrheology of complex materials. This is achieved by following a linear path that starts from a narrative introduction of the "nature of light," followed by a rigorous description of the fundamental equations governing the propagation of light through matter. Moreover, some of the many possible instrumental configurations are presented, especially those that better adapt to perform microrheology measurements. In order to better appreciate the microrheological methods with optical tweezers explored in this book, informative introductions to the basic concepts of linear rheology, statistical mechanics, and the most popular microrheology techniques are also given. Furthermore, an enlightening prologue to the general applications of optical tweezers different from rheological purposes is provided at the end of the book.

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