The goto resource for microscopists on biological applications of field emission gun scanning electron microscopy (FEGSEM)

The evolution of scanning electron microscopy technologies and capability over the past few years has revolutionized the biological imaging capabilities of the microscopegiving it the capability to examine surface structures of cellular membranes to reveal the organization of individual proteins across a membrane bilayer and the arrangement of cell cytoskeleton at a nm scale. Most notable are their improvements for field emission scanning electron microscopy (FEGSEM), which when combined with cryo-preparation techniques, has provided insight into a wide range of biological questions including the functionality of bacteria and viruses. This full-colour, must-have book for microscopists traces the development of the biological field emission scanning electron microscopy (FEGSEM) and highlights its current value in biological research as well as its future worth.

Biological Field Emission Scanning Electron Microscopy highlights the present capability of the technique and informs the wider biological science community of its application in basic biological research. Starting with the theory and history of FEGSEM, the book offers chapters covering: operation (strengths and weakness, sample selection, handling, limitations, and preparation); Commercial developments and principals from the major FEGSEM manufacturers (Thermo Scientific, JEOL, HITACHI, ZEISS, Tescan); technical developments essential to bioFEGSEM; cryobio FEGSEM; cryo-FIB; FEGSEM digital-tomography; array tomography; public health research; mammalian cells and tissues; digital challenges (image collection, storage, and automated data analysis); and more.

An important part of the Wiley-Royal Microscopical Series,Biological Field Emission Scanning Electron Microscopy is an ideal general resource for experienced academic and industrial users of electron microscopyspecifically, those with a need to understand the application, limitations, and strengths of FEGSEM.

ROLAND A. FLECK, PHD, FRCPath, FRMS, is a Professor in Ultrastructural Imaging and Director of the Centre for Ultrastructural Imaging at King's College London. Having specialised in basic research into cellular injury at low temperatures and during cryo-preservation regimes he has developed specialist knowledge of freeze fracture/freeze etch preparation of tissues and wider cryo-microscopic techniques. As director of the Centre for Ultrastructural Imaging he supports advanced three dimensional studies of cells and tissues by both conventional room temperature and cryo electron microscopy. He is a visiting Professor of the Faculty of Health and Medical Sciences, University of Copenhagen and Professor of the UNESCO Chair in Cryobiology, National Academy of Sciences of Ukraine, Institute for Problems of Cryobiology, Kharkiv, Ukraine.

BRUNO M. HUMBEL, Dr. sc. nat. ETH, is head of the Imaging Section at the Okinawa Institute of Science and Technology, Onna son, Okinawa, Japan. He is awarded a research professorship at Juntendo University, Tokyo, Japan. He got his PhD at the Federal Institute of Technology, ETH, Zurich, Switzerland, with Prof. Hans Moor and Dr. Martin Müller, both pioneers in cryo-electron microscopy (high-pressure freezing, freeze-fracturing, freeze-substitution and low-temperature embedding, cryo-SEM, cryo-sectioning). His research focuses on sample preparation for optimal, life-like imaging of biological objects in the electron microscope. The main interests are preparation methods based on cryo-fixation applied in Cell Biology. From here, hybrid follow-up methods like freeze-substitution or freeze-fracturing are used. He is also involved in immunolabelling technology, e.g., ultra-small gold particles and has been working on techniques for correlative microscopy and volume microscopy for a couple of years. He teaches cryo-techniques and immunolabelling and correlative microscopy in international workshops and has professional affiliations with Zhejiang University, Hangzhou, People's Republic of China as a distinguished professor and co-director of the Center of Cryo-Electron Microscopy and with the Federal University of Minas Gerais, Belo Horizonte, Brazil, as a FAPEMIG visiting professor at the Centro de Microscopia da UFMG.

About the Editors xix

List of Contributors xxi

Foreword xxv

1 Scanning Electron Microscopy: Theory, History and Development of the Field Emission Scanning Electron Microscope 1David C. Joy

2 Akashi Seisakusho Ltd SEM Development 19721986 7Michael F. Hayles

3 Development of FE-SEM Technologies for Life Science Fields 25Mitsugu Sato, Mami Konomi, Ryuichiro Tamochi and Takeshi Ishikawa

4 A History of JEOL Field Emission Scanning Electron Microscopes with Reference to Biological Applications 53Kazumichi Ogura and Andrew Yarwood

5 TESCAN Approaches to Biological Field Emission Scanning Electron Microscopy 79Jaroslav Jiru¨e, Vratislav Ko¨tál and Bohumila Lencová

6 FEG-SEM for Large Volume 3D Structural Analysis in Life Sciences 103Ben Lich, Faysal Boughorbel, Pavel Potocek and Emine Korkmaz

7 ZEISS Scanning Electron Microscopes for Biological Applications 117Isabel Angert, Christian Böker, Martin Edelman, Stephan Hiller, Arno Merkle and Dirk Zeitler

8 SEM Cryo-Stages and Preparation Chambers 143Robert Morrison

9 CryoSEM Specimen Preparation Workflows from the Leica Microsystems Design Perspective 167Guenter P. Resch

10 Chemical Fixation 191Bruno M. Humbel, Heinz Schwarz, Erin M. Tranfield and Roland A. Fleck

11 A Brief Review of Cryobiology with Reference to Cryo Field Emission Scanning Electron Microscopy 223Roland A. Fleck, Eyal Shimoni and Bruno M. Humbel

12 High-Resolution Cryo-Scanning Electron Microscopy of Macromolecular Complexes 265Sebastian Tacke, Falk Lucas, Jeremy D. Woodward, Heinz Gross and Roger Wepf

13 FESEM in the Examination of Mammalian Cells and Tissues 299Andrew Forge, Anwen Bullen and Ruth Taylor

14 Public Health/Pharmaceutical Research Pathology and Infectious Disease 311Paul A. Gunning and Bärbel Hauröder

15 Field Emission Scanning Electron Microscopy in Cell Biology Featuring the Plant Cell Wall and Nuclear Envelope 343Martin W. Goldberg

16 Low-Voltage Scanning Electron Microscopy in Yeast Cells 363Masako Osumi

17 Field Emission Scanning Electron Microscopy in Food Research 385Johan Hazekamp and Marjolein van Ruijven

18 Cryo-FEGSEM in Biology 397Paul Walther

19 Preparation of Vitrified Cells for TEM by Cryo-FIB Microscopy 415Yoshiyuki Fukuda, Andrew Leis and Alexander Rigort

20 Environmental Scanning Electron Microscopy 439Rudolph Reimer, Dennis Eggert and Heinrich Hohenberg

21 Correlative Array Tomography 461Thomas Templier and Richard H.R. Hahnloser

22 The Automatic Tape Collection UltraMicrotome (ATUM) 485Anwen Bullen

23 SBEM Techniques 495Christel Genoud

24 FIB-SEM for Biomaterials 517Lucille A. Giannuzzi

25 New Opportunities for FIB/SEM EDX in Nanomedicine: Cancerogenesis Research 533Damjana Drobne, Sara Novak, Andreja Erman and Goran Dra?ic

26 FIB-SEM Tomography of Biological Samples: Explore the Life in 3D 545Caroline Kizilyaprak, Damien De Bellis, Willy Blanchard, Jean Daraspe and Bruno M. Humbel

27 Three-Dimensional Field-Emission Scanning Electron Microscopy as a Tool for Structural Biology 567J.D. Woodward and R.A. Wepf

28 Element Analysis in the FEGSEM: Application and Limitations for Biological Systems 589Alice Warley and Jeremy N. Skepper

29 Image and Resource Management in Microscopy in the Digital Age 611Patrick Schwarb, Anwen Bullen, Dean Flanders, Maria Marosvölgyi, Martyn Winn, Urs Gomez and Roland A. Fleck

30 Part 1: Optimizing the Image Output: Tuning the SEM Parameters for the Best Photographic Results 625Oliver Meckes and Nicole Ottawa

31 A Synoptic View on Microstructure: Multi-Detector Colour Imaging, nanoflight® 659Stefan Diller

Index 679