Cell Imaging Techniques: Methods and Protocols

552 Pages·2013·10.101 MB·English

by  Douglas W. Cromey (auth.), Douglas J. Taatjes, Jürgen Roth (eds.)

Methods in
Molecular Biology 931
Douglas J. Taatjes
Jürgen Roth Editors
Cell Imaging
Techniques
Methods and Protocols
Second Edition
M M B ™
ETHODS IN OLECULAR IOLOGY
Series Editor
John M. Walker
School of Life Sciences
University of Hertfordshire
Hat fi eld, Hertfordshire, AL10 9AB, UK
For further volumes:
http://www.springer.com/series/7651

Cell Imaging Techniques
Methods and Protocols
Second Edition
Edited by
Douglas J. Taatjes
Department of Pathology and Microscopy Imaging Center, College of Medicine,
University of Vermont, Burlington, VT, USA
Jürgen Roth
Department of Integrated OMICS for Biomedical Science, WCU Program of Yonsei
University Graduate School, Yonsei University, Seoul, Korea
Editors
Douglas J. Taatjes Jürgen Roth
Department of Pathology and Microscopy Department of Integrated OMICS
Imaging Center for Biomedical Science
College of Medicine WCU Program of Yonsei University
University of Vermont Graduate School
Burlington, VT, USA Yonsei University
Seoul, Korea
ISSN 1064-3745 ISSN 1940-6029 (electronic)
ISBN 978-1-62703-055-7 ISBN 978-1-62703-056-4 (eBook)
DOI 10.1007/978-1-62703-056-4
Springer New York Heidelberg Dordrecht London
Library of Congress Control Number: 2012946194
© Springer Science+Business Media, LLC 2013
This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is
concerned, specifi cally the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction
on microfi lms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation,
computer software, or by similar or dissimilar methodology now known or hereafter developed. Exempted from this
legal reservation are brief excerpts in connection with reviews or scholarly analysis or material supplied specifi cally for
the purpose of being entered and executed on a computer system, for exclusive use by the purchaser of the work.
Duplication of this publication or parts thereof is permitted only under the provisions of the Copyright Law of the
Publisher’s location, in its current version, and permission for use must always be obtained from Springer. Permissions
for use may be obtained through RightsLink at theCopyright Clearance Center. Violations are liable to prosecution
under the respective Copyright Law.
The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not
imply, even in the absence of a specifi c statement, that such names are exempt from the relevant protective laws and
regulations and therefore free for general use.
While the advice and information in this book are believed to be true and accurate at the date of publication, neither
the authors nor the editors nor the publisher can accept any legal responsibility for any errors or omissions that may be
made. The publisher makes no warranty, express or implied, with respect to the material contained herein.
Printed on acid-free paper
Humana Press is a brand of Springer
Springer is part of Springer Science+Business Media (www.springer.com)
Dedication
This book is dedicated to Heidi, Joanna, and Andrea, for whom no microscope is required
to observe the joy you have brought to my life; and to my mother Jane for her abiding keen
interest in my research.
DJT

Preface
…by the help of Microscopes, there is nothing so small, as to escape our inquiry;
hence there is a new visible World discovered to the understanding.
Robert Hooke
Micrographia, 1665
In the fi rst edition of C ell Imaging Techniques (published in 2006), we sought to assemble
a volume of imaging protocols particularly useful for those working in a core imaging facil-
ity. We chose an eclectic representation of protocols, including techniques in fl uorescence
microscopy, confocal microscopy, atomic force microscopy, and electron microscopy,
amongst others. No one imaging mode was covered in detail, since individual volumes
devoted to such techniques have been and continue to be published, and serve as invaluable
reference sources for those seeking detailed protocols on a single imaging modality. In this
second edition of Cell Imaging Techniques , we have reevaluated the state of microscopy-
based imaging and have assembled a completely revised and expanded book. Only three
updated chapters remain from the original 21, with 26 new chapters added. Familiar tech-
niques from the fi rst edition, such as confocal microscopy, transmission electron micros-
copy, atomic force microscopy, and laser microdissection, have once again been included,
but with a somewhat different focus. New chapters have been added to refl ect ongoing
advances in imaging protocols. For instance, chapters covering colocalization analysis of
fl uorescent probes, correlative light and electron microscopy, environmental scanning elec-
tron microscopy, light sheet microscopy, intravital microscopy, high-throughput micros-
copy, and stereological techniques have now been included, just to name a few. Moreover,
methods to image specifi c organelles, such as lipid droplets, peroxisomes, and mitochon-
dria, as well as cellular processes such as endocytosis and autophagy have been added to this
expanded second edition. Ultimately, however, the goal of this second edition of the book
remains the same as that of the fi rst: to provide an easily accessible volume of protocols to
be used with a variety of imaging-based equipment likely available in a core imaging facility.
We hope that by perusing this volume, investigators in academic, clinical, and industrial
settings may be prompted to utilize a variety of microscopy-based imaging systems in their
research, perhaps even ones they had not considered previously.
At fi rst glance, the absence of chapters on the new super-resolution techniques may
seem a glaring oversight for a volume titled C ell Imaging Techniques. However, given the
rapidly evolving nature of these techniques, as well as the multiple solutions developed for
sub-diffraction optical microscopy, we chose not to include them in this volume. We trust
that given the global popularity of these novel methods for advanced imaging in cell biol-
ogy, volumes dedicated to this specifi c imaging area will likely be forthcoming.
vii
viii Preface
We again would like to express our sincerest appreciation to all of the authors who
provided chapters for this volume; they were a pleasure to work with, providing state-of-
the-art protocols and reviews in a timely fashion, while cheerfully responding to all of our
queries. We would also like to thank Professor John Walker, editor of the Methods in
Molecular Biology series, for his invaluable input and insight in all facets of the formulation
of this book.
This is truly an exciting time to be involved in microscopy-based imaging, as new tech-
nologies continue to be introduced at break-neck speed. We hope you will take advantage
of these imaging advances, and that this volume of protocols and reviews will serve as a
bench-top companion on your journey!
Burlington, VT, USA Douglas J. Taatjes
Seoul, Korea Jürgen Roth
Contents
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xiii
1 Digital Images Are Data: And Should Be Treated as Such . . . . . . . . . . . . . . . . 1
Douglas W. Cromey
2 Epi-Fluorescence Microscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
Donna J. Webb and Claire M. Brown
3 Live-Cell Migration and Adhesion Turnover Assays. . . . . . . . . . . . . . . . . . . . . 61
J. Lacoste, K. Young, and Claire M. Brown
4 Multifluorescence Confocal Microscopy: Application
for a Quantitative Analysis of Hemostatic Proteins
in Human Venous Valves. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85
Winifred E. Trotman, Douglas J. Taatjes, and Edwin G. Bovill
5 Colocalization Analysis in Fluorescence Microscopy . . . . . . . . . . . . . . . . . . . . 97
Jeremy Adler and Ingela Parmryd
6 A Time-Lapse Imaging Assay to Study Nuclear Envelope Breakdown . . . . . . . 111
Sunita S. Shankaran, Douglas R. Mackay, and Katharine S. Ullman
7 Light Sheet Microscopy in Cell Biology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
Raju Tomer, Khaled Khairy, and Philipp J. Keller
8 Image-Based High-Throughput Screening
for Inhibitors of Angiogenesis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
Lasse Evensen, Wolfgang Link, and James B. Lorens
9 Intravital Microscopy to Image Membrane Trafficking in Live Rats . . . . . . . . . 153
Andrius Masedunskas, Monika Sramkova, Laura Parente,
and Roberto Weigert
10 Imaging Non-fluorescent Nanoparticles in Living Cells
with Wavelength-Dependent Differential Interference
Contrast Microscopy and Planar Illumination Microscopy. . . . . . . . . . . . . . . . 169
Wei Sun, Lehui Xiao, and Ning Fang
11 Laser Scanning Cytometry: Principles and Applications—An Update. . . . . . . . 187
Piotr Pozarowski, Elena Holden, and Zbigniew Darzynkiewicz
12 Laser Capture Microdissection for Protein
and NanoString RNA Analysis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213
Yelena Golubeva, Rosalba Salcedo, Claudius Mueller,
Lance A. Liotta, and Virginia Espina
ix