Electron Microscopy: Methods and Protocols (Methods in Molecular Biology (1117))

This third edition of __Electron Microscopy: Methods and Protocols__ expands upon the previous editions with current, detailed protocols on biological and molecular research techniques based on TEM and SEM as well as other closely related imaging and analytical methods. With new chapters on conventional and microwave assisted specimen, cryo-specimen preparation, negative staining and immunogold labelling techniques, DNA and RNA tracking using hybrization in TEM or Atomic Force Microscopy, TEM crystallography and cryo TEM 3D tomography, 3D tomography of resin embedded tissues using FIB-SEM, Correlative microscopy using fluorescence microscopy, confocal microscopy or immune labelling techniques for both TEM and FIB-SEM and Elemental and isotopic identification and their distribution in cells and tissues using TEM, SEM, Scanning Transmission Electron Microscopy (STEM), Secondary Ion Mass Spectrometry (SIMS) and Nano SIMS. Written in the highly successful __Methods in Molecular Biology__ series format, 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, __Electron Microscopy: Methods and Protocols, Third Edition__ provides the most up-to-date and essential information in electron microscopy techniques and methods provided in this edition will assist in advancing future molecular and biological research. Conventional specimen preparation techniques for transmission electron microscopy of cultured cells / John J. Bozzola Microwave-assisted processing and embedding for transmission electron microscopy / Paul Webster Processing plant tissues for ultrastructural study / John Kuo Staining sectioned biological specimens for transmission electron microscopy : conventional and En Bloc stains / E. Ann Ellis Metal shadowing for electron microscopy / Gregory M. Hendricks Freeze fracture and freeze etching / Douglas E. Chandler and William P. Sharp Conventional specimen preparation techniques for scanning electron microscopy of biological specimens / John J. Bozzola High-pressure freezing : current state and future prospects / Andres Kaech and Urs Ziegler Cryo-fixation by self-pressurized rapid freezing / Markus Grabenbauer, Hong-Mei Han, and Jan Huebinger Cryo-electron microscopy of vitreous sections / Petr Chlanda and Martin Sachse Negative staining and cryo-negative staining : applications in biology and medicine / J. Robin Harris and Sacha De Carlo Electron microscopy of microtubule cytoskeleton assembly in vitro / Margaret Coughlin, Aaron C. Groen, and Timothy J. Mitchison Cryosectioning fixed and cryoprotected biological material for immunocytochemistry / Paul Webster and Alexandre Webster Analysis of specificity in immunoelectron microscopy / Christian Hacker and John M. Lucocq Cryo-electron microscopy of membrane proteins / Kenneth N. Goldie [and five others] Tracking DNA and RNA sequences at high resolution / Dušan Cmarko, Anna Ligasová, and Karel Koberna Visualization of DNA and protein-DNA complexes with atomic force microscopy / Yuri L. Lyubchenko, Alexander A. Gall, and Luda S. Shlyakhtenko Biological applications of phase-contrast electron microscopy / Kuniaki Nagayama Single particle cryo-electron microscopy and 3-D reconstruction of viruses / Fei Guo and Wen Jiang Electron tomography for organelles, cells, and tissues / Wanzhong He and Yongning He Correlative light and electron microscopy : from live cell dynamic to 3D ultrastructure / Coralie Spiegelhalter, Jocelyn F. Laporte, and Yannick Schwab Nanometer-resolution fluorescence electron microscopy (Nano-EM) in cultured cells / Shigeki Watanabe [and nine others] Correlative fluorescence and electron microscopy of quantum dot labeled proteins on whole cells in liquid / Diana B. Peckys, Madeline J. Dukes, and Niels de Jonge FIB-SEM tomography in biology / Caroline Kizilyaprak [and three others] Correlative light and electron microscopy using immunolabeled sections / Heinz Schwarz and Bruno M. Humbel Correlative 3D imaging : CLSM and FIB-SEM tomography using high-pressure frozen, freeze-substituted biological samples / Miriam S. Lucas [and four others] Three-dimensional imaging of adherent cells using FIB/SEM and STEM / Clarissa Villinger [and six others] X-ray microanalysis in the scanning electron microscope / Godfried M. Roomans and Anca Dragomir Application of SEM and EDX in studying biomineralization in plant tissues / Honghua He and Yaowanuj Kirilak Freeze stabilization and cryopreparation technique for visualizing the water distribution in woody tissues by X-ray imaging and cryo-scanning electron microscopy / Yasuhiro Utsumi and Yuzou Sano Biological applications of energy-filtered TEM / Martin Saunders and Jeremy A. Shaw Secondary ion mass spectrometry imaging of biological cells and tissues / Nicholas P. Lockyer Elemental and isotopic imaging of biological samples using nanoSIMS / Matt R. Kilburn and Peta L. Clode 3D chemical mapping : application of scanning transmission (Soft) x-ray microscopy (STXM) in combination with angle-scan tomography in bio-, geo-, and environmental sciences / Martin Obst and Gregor Schmid. Annotation This third edition of "Electron Microscopy: Methods and Protocols "expands upon the previous editions with current, detailed protocols on biological and molecular research techniques based on TEM and SEM as well as other closely related imaging and analytical methods. With new chapters on conventional and microwave assisted specimen, cryo-specimen preparation, negative staining and immunogold labelling techniques, DNA and RNA tracking using hybrization in TEM or Atomic Force Microscopy, TEM crystallography and cryo TEM 3D tomography, 3D tomography of resin embedded tissues using FIB-SEM, Correlative microscopy using fluorescence microscopy, confocal microscopy or immune labelling techniques for both TEM and FIB-SEM and Elemental and isotopic identification and their distribution in cells and tissues using TEM, SEM, Scanning Transmission Electron Microscopy (STEM), Secondary Ion Mass Spectrometry (SIMS) and Nano SIMS. Written in the highly successful "Methods in Molecular Biology "series format, 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, "Electron Microscopy: Methods and Protocols, Third Edition" provides the most up-to-date and essential information in electron microscopy techniques and methods provided in this edition will assist in advancing future molecular and biological research Comprises two major areas of electron microscopy-transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The TEM area covers several key techniques, including: conventional specimen preparation methods for cultured cells and biomedical and plant tissues; cryospecimen preparation by high-pressure freezing and cryoultramicrotomy negative staining and immunogold labeling techniques; and TEM crystallography and cryo-TEM tomography. The SEM area similarly attends to conventional-, variable pressure-, environmental-, and cryoscanning microscopy techniques, as well as the application of X-ray microanalysis. Protocols for the application of X-ray microanalysis to SEM and mass spectrometry conclude the volume. Front Matter....Pages i-xviii Back Matter....Pages 1-19 ....Pages 21-37