This book encompasses the full breadth of the super-resolution imaging field, representing modern techniques that exceed the traditional diffraction limit, thereby opening up new applications in biomedicine. It shows readers how to use the new tools to increase resolution in sub-nanometer-scale images of living cells and tissue, which leads to new information about molecules, pathways and dynamics. The book highlights the advantages and disadvantages of the techniques, and gives state-of-the-art examples of applications using microscopes currently available on the market. It covers key techniques such as stimulated emission depletion (STED), structured illumination microscopy (SSIM), photoactivated localization microscopy (PALM), and stochastic optical reconstruction microscopy (STORM). It will be a useful reference for biomedical researchers who want to work with super-resolution imaging, learn the proper technique for their application, and simultaneously obtain a solid footing in other techniques.
Table of Contents
Overview of Super-Resolution Technologies. Introduction to the Main Techniques - (F)PALM/(d)STORM, STED, SIM. Super-Resolution Imaging of Single Molecules. Image Processing for Super-Resolution. Super-Resolution Fluorescent Probes. The Methods. Principles and Techniques for STED; SIM/SSIM; SOFI. Single-Molecule Localization Microscopy - PALM, (F)PALM, (d)STORM): Principles, Probes, and Algorithms. Principles and Techniques for SPDM; GSDIM. Applications in Biology and Medicine. Synaptic Protein Distributions Studied with (d)STORM/(F)PALM. Studies of Chromosomal Organization at Super Resolution. Nanoscale Organization of RNA Polymerase Using PALM. Super-Resolution Microscopy of Protein Machines in Bacteria. Super-Resolution Microscopy of Living Biological Nanostructures. SIM for the Study of Mitochondrial Dynamics in 3D. Neurobiological Applications Using STED. STED in HIV Research. Super-Resolution Imaging in Cardiovascular Research. Quantitative Super-Resolution Imaging of Proteins.
Alberto Diaspro, Ph.D, is a professor of applied physics and head of the Nanophysics Unit of the Italian Institute of Technology. He also founded the Laboratory for Advanced Microscopy, Bioimaging, and Spectroscopy (LAMBS).
Marc A.M.J. van Zandvoort, Ph.D, is the leader of the Two-Photon Core Facility in Aachen and head of the Advanced Microscopy Unit in the Department of Genetics and Cell Biology at Maastricht University, Netherlands.