Description
For more than a century, microscopy has been a centerpiece of extraordinary discoveries in biology. Along the way, remarkable imaging tools have been developed allowing scientists to dissect the complexity of cellular processes at the nano length molecular scales. Nanoimaging: Methods and Protocols presents a diverse collection of microscopy techniques and methodologies that provides guidance to successfully image cellular molecular complexes at nanometer spatial resolution. The book’s four parts cover: (1) light microscopy techniques with a special emphasis on methods that go beyond the classic diffraction-limited imaging; (2) electron microscopy techniques for high-resolution imaging of molecules, cells and tissues, in both two and three dimensions; (3) scanning probe microscopy techniques for imaging and probing macromolecular complexes and membrane surface topography; and (4) complementary techniques on correlative microscopy, soft x-ray tomography and secondary ion mass spectrometry imaging. Written in the successful format of the Methods in Molecular Biology(TM) series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Nanoimaging: Methods and Protocols highlights many of the most exciting possibilities in microscopy for the investigation of biological structures at the nano length molecular scales. 1. Introduction: Nanoimaging Techniques in Biology Alioscka A. Sousa and Michael J. Kruhlak Part I. Light Microscopy 2. Live-Cell Imaging of Vesicle Trafficking and Divalent Metal Ions by Total Internal Reflection Fluorescence (TIRF) Microscopy Merewyn K. Loder, Takashi Tsuboi, and Guy A. Rutter 3. 4Pi Microscopy Roman Schmidt, Johann Engelhardt, and Marion Lang 4. Fluorescence in situ Hybridization Applications for Super-Resolution 3D Structured Illumination Microscopy Yolanda Markaki, Daniel Smeets, Marion Cremer, and Lothar Schermelleh 5. Two-Color STED Imaging of Synapses in Living Brain Slices Jan Tnnesen, and U. Valentin Ngerl 6. Super-Resolution Imaging by Localization Microscopy Dylan M. Owen, Astrid Magenau, David Williamson, and Katharina Gaus 7. High-Content Super-Resolution Imaging of Live Cell by uPAINT Grgory Giannone, Eric Hosy, Jean-Baptiste Sibarita, Daniel Choquet, and Laurent Cognet 8. Super-Resolution Fluorescence Imaging with Blink Microscopy Christian Steinhauer, Michelle S. Itano, and Philip Tinnefeld 9. Photoswitchable Fluorophores for Single-Molecule Localization Microscopy Kieran Finan, Benjamin Flottmann, and Mike Heilemann 10. Single-Molecule Tracking of mRNA in Living Cells Mai Yamagishi, Yoshitaka Shirasaki, and Takashi Funatsu Part II. Electron Microscopy 11. Semiautomatic, High-Throughput, High-Resolution Protocol for Three-Dimensional Reconstruction of Single Particles in Electron Microscopy Carlos Oscar Sorzano, J. M. de la Rosa Trevn, J. Otn, J. J. Vega, J. Cuenca, A. Zaldvar-Peraza, J. Gmez-Blanco, J. Vargas, A. Quintana, R. Marabini, and J. M. Carazo 12. Mass Mapping of Amyloid Fibrils in the Electron Microscope using STEM Imaging Alioscka A. Sousa and Richard D. Leapman 13. Elemental Mapping by Electron Energy Loss Spectroscopy in Biology Maria A. Aronova and Richard D. Leapman 14. Cellular Nanoimaging by Cryo Electron Tomography Roman I. Koning and Abraham J. Koster 15. Large-Volume Reconstruction of Brain Tissue from High-Resolution Serial Section Images Acquired by SEM-Based Scanning Transmission Electron Microscopy Masaaki Kuwajima, John M. Mendenhall, and Kristen M. Harris 16. 3D Imaging of Cells and Tissues by Focused Ion Beam/Scanning Electron Microscopy (FIB/SEM) Damjana Drobne 17. Preparation of Gold Nanocluster Bioconjugates for Electron Microscopy Christine L. Heinecke and Christopher J. Ackerson Part III. Scanning Probe Microscopy 18. Atomic Force Microscopy Imaging of Macromolecular Complexes S. Santos, D.J. Billingsley, and N.H. Thomson 19. Imaging of Transmembrane Proteins Directly Incorporated Within Supported Lipid Bilayers Using Atomic Force Microscopy Daniel Levy and Pierre-Emmanuel Milhiet 20. Functional AFM Imaging of Cellular Membranes Using Functionalized Tips Lilia A. Chtcheglova and Peter Hinterdorfer 21. Near-Field Scanning Optical Microscopy for High Resolution Membrane Studies Heath A. Huckabay, Kevin P. Armendariz, William H. Newhart, Sarah M. Wildgen, and Robert C. Dunn Part IV. Complementary Techniques 22. Correlative Fluorescence and EFTEM Imaging of the Organized Components of the Mammalian Nucleus Michael J. Kruhlak 23. High Data Output Method for 3-D Correlative Light-Electron Microscopy Using Ultrathin Cryosections Katia Cortese, Giuseppe Vicidomini, Maria C. Gagliani, Patrizia Boccacci, Alberto Diaspro, and Carlo Tacchetti 24. Correlative Optical and Scanning Probe Microscopies for Mapping Interactions at Membranes Christopher M. Yip 25. Nanoimaging Cells Using Soft X-Ray Tomography Dilworth Y. Parkinson, Lindsay R. Epperly, Gerry McDermott, Mark A. Le Gros, Rosanne M. Boudreau, and Carolyn A. Larabell 26. Secondary Ion Mass Spectrometry Imaging of Biological Membranes at High Spatial Resolution Haley A. Klitzing, Peter K. Weber, and Mary L. Kraft
