Multimodal And Nanoscale Optical Microscopy

Optical microscopy is developing into nanoscopy and multimodal microscopy the better to decipher the functioning mechanisms in living systems, and investigating biological specimens at molecular level using fluorescence as a mechanism of contrast. Results have demonstrated the potential to provide information at the Angstrom level. Other optical methods now offer more in terms of spatial and temporal resolution, making it possible to study the delicate and complex relationship between structure and function in cells. Modern optical microscopes also use the decisive advantage provided by artificial intelligence algorithms. All in all, a rapidly changing field with an increasing number of questions to be answered. This book contains the lectures presented as Course 210 of the prestigious International School of Physics Enrico Fermi, Multimodal and Nanoscale Optical Microscopy, held in Varenna, Italy from 11 to 16 July 2021. Topics covered include fluorescence; linear and nonlinear microscopy; label-free with Mueller matrix and Brillouin microscopy; F-methods such as FRAP, FLIM, FRET and FCS; super-resolution, phototoxicity and photodamage; optical and magnetic trapping; image formation; and bioimage analysis, among others. The book also includes selected contributions of a number of young researchers. Packed with novel ideas, new instruments, challenging theoretical approaches and amazing applications, the book will be of interest to all those working in the field of multimodal and nanoscale optical microscopy.

"Alberto Diaspro has been choreographing light’s dance for over 20 years, and in Nanoscopy and Multidimensional Optical Fluorescence Microscopy, he has assembled a diverse group of experts to explain the methods they use to coax light to reveal biology’s secrets." — From the Foreword by Daniel Evanko, editor, Nature Methods Nanoscopy and Multidimensional Optical Fluorescence Microscopy demonstrates that the boundaries between sciences do blur at the bottom, especially those that might separate the optical work of physicists and the cellular work of microbiologists. In 18 chapters written by pioneering researchers, this work offers the first comprehensive and current documentation of the cutting-edge research being accomplished in a wide range of photonic devices with revolutionary application. The highlight of the book is its coverage of optical nanoscopy and super-resolution microscopy. The rapid advances in this area over the past few years offer researchers in both photonics and molecular biologya wealth of accomplishment upon which they can build. Offering a complete treatment of this emerging field, this volume: Describes how scientists have exploited the properties of light and its fluorophore partners to overcome the resolution limit of conventional light microscopy Delves into recent ways to minimize the photobleaching that has long hampered many methods including those that have the potential to capture previously unobtainable information on the movements of single molecules Discusses the principles, benefits, and implementation of fluorescence correlation spectroscopy and related methods, which simplifies analysis by limiting light to stationary focal points in a sample Considers the most basic as well as emerging methods for improving three-dimensional optical sectioning microscopy Reviews the basics of FRET (fluorescence resonance energy transfer) and considers its new use for investigating protein complexes The text also introduces those emerging nonfluorescence microscopy methods that can actually exert mechanical forces to trap and move a variety of objects ranging from beads to living cells and cellular organelles. Combining this technique with fluorescence microscopy provides an unparalleled ability to manipulate and visualize biological samples. In the half-century since Richard Feynman challenged scientists to come up with the tools to investigate and manipulate our world at the nanoscale, we have succeeded in placing tools in the hands of biophysicists that are leading to major breakthroughs in our understanding of life and our ability to diagnose, treat, and prevent many challenges to human health. This book reflects what has been accomplished to date while pointing the way to what still needs to be done.

"This groundbreaking book focuses on near-field microscopy which has opened up optical processes at the nanoscale for direct inspection. Further, it explores the emerging area of nano-optics which promises to make possible optical microscopy with true nanometer resolution. This frontline resource helps you achieve high resolution optical imaging of biological species and functional materials. You also find guidance in the imaging of optical device operation and new nanophotonics functionalities"--EBL.

The story of microscopy over the years is one of wonder, revelation, and even love. What better words could there be to describe the amazing things that we have been able to see, learn and accomplish thanks to the progress made in this field? A love story between a pieace of glass and the rainbow with an original soundtrack mad of poetry and music. From Galilei’s initial foray into basic optical microscopy, including the Camillo Golgi and Giuliano Toraldo di Francia lessons, to such later developments as time-resolved microscopy, multi-photon microscopy and three-dimensional microscopy to innovations such as optical nanoscopy, bioimaging and super resolution imaging, the book seeks to take the reader, be they scientist or layperson, on a journey through the evolution of the microscope and its many uses, including in the field of medicine. The author uses visible light as a through-line to unite the various chapters, as well as using fluorescence as a touchpoint from which to map the changes in the science, a significant choice, as it, along with label-free approaches and the addition of artificial intelligence, form the natural environment for development of the modern multi-messenger microscope towards bioimaging at the nanoscale.

This book describes developments in the field of super-resolution fluorescence microscopy or nanoscopy. In 11 chapters, distinguished scientists and leaders in their respective fields describe different nanoscopy approaches, various labeling technologies, and concrete applications. The topics covered include the principles and applications of the most popular nanoscopy techniques STED and (f)PALM/STORM, along with advances brought about by fluorescent proteins and organic dyes optimized for fluorescence nanoscopy. Furthermore, the photophysics of fluorescent labels is addressed, specifically for improving their photoswitching capabilities. Important applications are also discussed, such as the tracking and counting of molecules to determine acting forces in cells, and quantitative cellular imaging, respectively, as well as the mapping of chemical reaction centers at the nano-scale. The 2014 Chemistry Nobel Prize® was awarded for the ground-breaking developments of super-resolved fluorescence microscopy. In this book, which was co-edited by one of the prize winners, readers will find the most recent developments in this field.

In the last decade, fluorescence microscopy has evolved from a classical “retrospective” microscopy approach into an advanced imaging technique that allows the observation of cellular activities in living cells with increased resolution and dimensions. A bright new future has arrived as the nano era has placed a whole new array of tools in the hands of biophysicists who are keen to go deeper into the intricacies of how biological systems work. Following an introduction to the complex world of optical microscopy, this book covers topics such as the concept of white confocal, nonlinear optical microscopy, fluctuation spectroscopies, site-specific labeling of proteins in living cells, imaging molecular physiology using nanosensors, measuring molecular dynamics, muscle braking and stem cell differentiation.

Microscopy is at the forefront of multidisciplinary research. It was developed by physicists, made specific by chemists, and applied by biologists and doctors to better understand how the human body works. For this very reason, the field has been revolutionized in past decades. The objective of Optical Nanoscopy and Novel Microscopy Techniques is to choose some of those revolutionary ideas and serve a general audience from broad disciplines to achieve a fundamental understanding of these technologies and to better apply them in their daily research. The book begins with coverage of super-resolution optical microscopy, which discusses targeted modulation such as STED and SIM or localization methods such as PALM. It then discusses novel development of fluorescent probes, such as organic small-molecule probes, fluorescent proteins, and inorganic labels such as quantum dots. Finally, it describes advanced optical microscopy, such as fluorescence lifetime imaging, fiber optic microscopy, scanning ion conductance microscopy, and the joining of optics and acoustics—photoacoustic microscopy. Following each chapter, a detailed list of references is provided. Problems at the end of each chapter are also included.