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