Live Cell Imaging

by Goldman, Robert D.; Spector, David L.

• ISBN13: 9780879696832
• ISBN10: 0879696834
• Edition: DVD
• Format: Paperback
• Copyright: 2004-11-30
• Publisher: Cold Spring Harbor Laboratory Pr

Summary

Recent advances in imaging technology reveal, in real time and great detail, critical changes in living cells and organisms. This manual is a compendium of emerging techniques, organized into two parts: specific methods such as fluorescent labeling, and delivery and detection of labeled molecules in cells; and experimental approaches ranging from the detection of single molecules to the study of dynamic processes in organelles, organs, and whole animals. Although presented primarily as a laboratory manual, the book includes introductory and background material and could be used as a textbook in advanced courses. It also includes a DVD containing movies of living cells in action, created by investigators using the imaging techniques discussed in the book. The editors, David Spector and Robert Goldman, whose previous book was Cells: A Laboratory Manual,are highly respected investigators who have taught microscopy courses at Cold Spring Harbor Laboratory, the Marine Biology Laboratory at Woods Hole, and Northwestern University.

Table of Contents

DVD Contents	xi
Preface	xv
SECTION 1: DETECTION AND APPROACHES TO LIVE CELL IMAGING	1	(326)
Fluorescent Protein Tracking and Detection	3	(22)
M. Rizzo
D. Piston
Constructing and Expressing GFP Fusion Proteins	25	(8)
D. Spector
R. Goldman
Viral Vectors for Introduction of GPF	33	(18)
R. Lansford
Gene Delivery by Direct Injection and Facilitation of Expression by Mechanical Stretch	51	(16)
D. Dean
Microinjection of Fluorophore-labeled Proteins	67	(20)
Y. Komarova
J. Peloquin
G. Borisy
CCD Cameras for Fluorescence Imaging of Living Cells	87	(14)
P. Tran
Photobleaching Techniques to Study Mobility and Molecular Dynamics of Proteins in Live Cells: FRAP, iFRAP, and FLIP	101	(26)
G. Rabut
J. Ellenberg
FRET and Fluorescence Lifetime Imaging Microscopy	127	(18)
M. Tramier
D. Sanvitto
V. Emiliani
C. Durieux
M. Coppey-Moisan
Monitoring Protein Dynamics Using FRET-based Biosensors	145	(14)
T. Chew
R. Chisholm
Application of Light-directed Activation of Caged Biomolecules and CALI to Problems in Cell Motility	159	(18)
D. Humphrey
Z. Rajfur
B. Imperiali
G. Marriott
P. Roy
K. Jacobson
Photoactivation-based Labeling and In Vivo Tracking of RNA Molecules in the Nucleus	177	(10)
J. Politz
R. Tuft
T. Pederson
Fluorescent Speckle Microscopy of Cytoskeletal Dynamics in Living Cells	187	(18)
T. Wittman
R. Littlefield
C. Waterman-Storer
Polarization Microscopy with the LC-PolScope	205	(34)
R. Oldenbourg
Confocal Microscopy, Deconvolution, and Structured Illumination Methods	239	(42)
J. Murray
Multiphoton and Multispectral Laser-scanning Microscopy	281	(22)
M. Dickinson
Analyzing Live Cell Data and Tracking Dynamic Movements	303	(24)
W. Tvarusko
J. Mattes
R. Eils
SECTION 2: IMAGING OF LIVE CELLS AND ORGANISMS	327	(290)
In Vivo Imaging of Mammalian Cells	329	(16)
J. Swedlow
P. Andrews
M. Platani
A Sealed Preparation for Long-term Observations of Cultured Cells	345	(6)
G. Sluder
J. Nordberg
F. Miller
E. Hinchcliffe
Live Cell Imaging of Yeast	351	(20)
D. Rines
D. Thomann
J. Dorn
P. Goodwin
P. Sorger
Live Imaging of Caenorhabditis elegans	371	(14)
B. Podbilewicz
Y. Gruenbaum
Time-lapse Cinematography in Living Drosophila Tissues	385	(24)
I. Davis
R. Parton
Single-cell Imaging in Animal Tumors In Vivo	409	(14)
J. Wyckoff
J. Segall
J. Condeelis
Long-term, High-resolution Imaging in the Neocortex In Vivo	423	(12)
B. Chen
J. Trachtenberg
A. Holtmaat
K. Svoboda
Intravital Microscopy of Normal and Diseased Tissues in the Mouse	435	(32)
R. Jain
E. Brown
L. Munn
D. Fukumura
Development of Mammalian Cell Lines with lac Operator-tagged Chromosomes	467	(16)
Y. Strukov
A. Belmont
Imaging Gene Expression in Living Cells	483	(8)
S. Janicki
D. Spector
Studying Mitosis in Cultured Mammalian Cells	491	(12)
P. Wadsworth
Imaging Hoechst-33342-labeled Chromosomes and Fluorescent Proteins During the Cell Cycle	503	(10)
T. Haraguchi
Y. Hiraoka
Imaging the Actin Cytoskeleton	513	(10)
A. Matus
V. Biou
H. Brinkhaus
M. Roelandse
Imaging Intermediate Filament Proteins in Living Cells	523	(14)
E. Kuczmarski
R. Goldman
Methods for Expressing and Analyzing GFP-Tubulin and GFP-Microtubule-associated Proteins	537	(18)
H. Goodson
P. Wadsworth
Imaging of Organelle Membrane Systems and Membrane Traffic in Living Cells	555	(14)
J. Lippincott-Schwartz
E. Snapp
Imaging Live Cells Under Mechanical Stress	569	(16)
B. Helmke
P. Davies
Imaging Single Molecules Using Total Internal Reflection Fluorescence Microscopy	585	(18)
N. Stuurman
R. Vale
Visualization and Quantification of Single RNA Molecules in Living Cells	603	(14)
Y. Shav-Tal
S. Shenoy
R. Singer
Appendix 1: Cautions	617	(8)
Index	625