© 2005 • 854 pp., illus., appendices, index
Paperback • $167 • ISBN  978-087969692-4

This title also available in: Hardcover

As imaging studies have continued to expand in scope and sophistication, this new edition of the highly successful and well–received Imaging Neurons: A Laboratory Manual has expanded to include development, with over twenty new chapters on such topics as MRI microscopy, imaging early developmental events, and labeling single neurons. Chapters on FRET, FCS/ICS, FRAP, hyperresolution microscopy, single molecule imaging, imaging with quantum dots, and imaging gene expression are included. With over forty full chapters, the manual also includes over forty sections of protocols for imaging techniques.

  “The book originated at the Cold Spring Harbor summer course on imaging of neuronal structure and function of the nervous system. Course directors Yuste and Konnerth did a great service to the neuroscience community by compiling a manual that contains basic imaging principles as well as a long list of protocols. Though the concepts underlying imaging techniques are familiar to most people using a microscope, the devil is in the details. Why isn’t my dye going into solution? How can I prevent my sample from bleaching faster than I can image? What camera do I need? It is in the answers to these sorts of questions where this manual succeeds by providing a practical guide for researchers and students who want to pursue imaging experiments.”
      —Nature Neuroscience

  “Directed towards students and researchers whose work involves imaging in neuroscience, developmental biology, and cell biology, this new edition is designed for use at the setup or bench. The emphasis is on the practical techniques and procedures that are used to optically image a variety of preparations.  The text is augmented with an extensive section on microscopy principles and instrumentation, which could serve the researcher who works in biological imaging.  Since the majority of the chapters are composed of protocols, techniques, and detailed procedures, I think that this book can best be used as a laboratory manual to accompany an undergraduate or graduate course on optical imaging with emphasis on neurobiology and developmental biology.

...the production quality is very good; the figures (many in color) and line drawings are very sharp, and they augment the text. The book includes a comprehensive index as well.”
      —Journal of Biomedical Optics

  “The book offers a broad coverage of many of the most important aspects of imaging technologies widely used in the neuroscience and developmental biology practice and that have revolutionized research in many areas of biology.

The editors have made sincere efforts to diminish the gap between theoretical knowledge and practical setup of experiments by preparing this manual with wider objectives and indexes with focused application or experiment in mind. Given examples expose the strengths and weaknesses of the method for a given situation, aimed at helping readers choose appropriate protocols and utilize them for their own laboratory conditions...

The book comes at an appropriate time since it aims at providing professional technical education by, as editors claim themselves, ‘teaching the investigators often tricky art of applying the novel microscopy approaches to problems in cellular and developmental neurobiology’. Many manuals have been written on imaging in neuroscience, but few have applied it across such a wide swathe of field.  The book is well produced and errors are minor. I recommend this work as a major contribution for advanced undergraduates, postgraduates, basic neuroscientists and clinicians interested in imaging in neuroscience and developmental or cell biology.”
      —Journal of Chemical Neuroanatomy

SECTION 1: BASIC IMAGING

Chapter 1: Maintaining Live Cells and Tissue Slices in the Imaging Setup
M.E. Dailey, G.S. Marrs, and D. Kurpius

Chapter 2: Video Microscopy, Video Cameras, and Image Enhancement
M. Oshiro, L.A. Moomaw, and E. Keller

Chapter 3: The Application of Scientific–grade CCD Cameras to Biological Imaging
M. Christenson

Chapter 4: A Practical Guide: Differential Interference Contrast Imaging of Living Cells
N.E. Ziv and J. Schiller

Chapter 5: A Practical Guide: Infrared Video Microscopy
H.–U. Dodt, K. Becker, and W. Zieglgänsberger

Chapter 6: Confocal Microscopy: Principles and Practice
A. Fine

Chapter 7: Principles of Multiphoton–excitation Fluorescence Microscopy
W. Denk

Chapter 8: Two–Photon Microscopy for 4D Imaging of Living Neurons
S.M. Potter

Chapter 9: A Practical Guide: Building a Two–Photon Laser–scanning Microscope
J. Mertz

Chapter 10: A Practical Guide: How to Build a Two–Photon Microscope Using a Confocal Scan Head
V. Nikolenko and R. Yuste

SECTION 2: IMAGING DEVELOPMENT

Chapter 11: In Vivo Electroporation during Embryogenesis
C.E. Krull, R. McLennan, S. O’Connell, Y. Chen, and P.A. Trainor

Chapter 12: A Practical Guide: In Vivo Electroporation of Neurons
S.R. Price

Chapter 13: A Practical Guide: Single–Neuron Labeling Using Genetic Methods
L. Luo

Chapter 14: A Practical Guide: Ballistic Delivery of Dyes for Structural and Functional Studies of the Nervous System
W.–B. Gan, J. Grutzendler, R.O. Wong, and J.W. Lichtman

Chapter 15: A Practical Guide: Imaging Embryonic Development in Caenorhabditis elegans
W.A. Mohler and A.B. Isaacson

Chapter 16: Visualizing Morphogenesis in Frog Embryos
L.A. Davidson and J.B. Wallingford

Chapter 17: In Vivo Imaging of Synaptogenesis in the Embryonic Zebrafish
J.D. Jontes and S.J Smith

Chapter 18: A Practical Guide: In Ovo Imaging of Avian Embryogenesis
P.M. Kulesa and S.E. Fraser

Chapter 19: A Practical Guide: Time–lapse Imaging of the Formation of the Chick Peripheral Nervous System
J.C. Kasemeier, F. Lefcort, S.E. Fraser, and P.M. Kulesa

Chapter 20: Imaging Mouse Embryonic Development
E.A.V. Jones, A.–K. Hadjantonakis, and M.E. Dickinson

Chapter 21: Imaging the Developing Retina
C. Lohmann, J.S. Mumm, J. Morgan, L. Godinho, E. Schroeter, R. Stacy, W.T. Wong, D. Oakley, and R.O.L. Wong

Chapter 22: A Practical Guide: Long–term Two–Photon Transcranial Imaging of Synaptic Structures in the Living Brain
J. Grutzendler and W.–B. Gan

Chapter 23: In Vivo Time–lapse Imaging of Neuronal Development
E.S. Ruthazer, K. Haas, A. Javaherian, K. Jensen, W.C. Sin, and H.T. Cline

Chapter 24: Optical Projection Tomography: Imaging 3D Organ Shapes and Gene Expression Patterns in Whole Vertebrate Embryos
J. Sharpe

Chapter 25: Imaging the Development of the Neuromuscular Junction
M.K. Walsh and J.W. Lichtman

Chapter 26: A Practical Guide: Imaging Synaptogenesis by Measuring Accumulation of Synaptic Proteins
C. Dean and P. Scheiffele

Chapter 27: A Practical Guide: Imaging Retinotectal Synaptic Connectivity
S. Cohen–Corey

Chapter 28: A Practical Guide: Intrinsic Optical Imaging of Functional Map Development in Mammalian Visual Cortex
T. Bonhoeffer and M. Hübener

SECTION 3: CALCIUM IMAGING

Chapter 29: How Calcium Indicators Work
S.R. Adams

Chapter 30: Some Quantitative Aspects of Calcium Fluorimetry
E. Neher

Chapter 31: Calibration of Fluorescent Calcium Indicators
F. Helmchen

Chapter 32: A Single–Compartment Model of Calcium Dynamics in Nerve Terminals and Dendrites
F. Helmchen and D.W. Tank

Chapter 33: A Practical Guide: Dye Loading with Patch Pipettes
J. Eilers and A. Konnerth

Chapter 34: A Practical Guide: Calcium Imaging of the Retina
C. Lohmann, J. Demas, J.L. Morgan, and R.O.L. Wong

Chapter 35: Calcium Imaging of Identified Astrocytes in Hippocampal Slices
J. Kang, G. Arcuino, and M. Nedergaard

Chapter 36: Imaging Microscopic Calcium Signals in Excitable Cells
M.B. Cannell, A.J.C. McMorland, and C. Soeller

Chapter 37: Monitoring Presynaptic Calcium Dynamics with Membrane–permeant Indicators
W.G. Regehr

Chapter 38: A Practical Guide: Two–Photon Calcium Imaging of Spines and Dendrites
J.H. Goldberg and R. Yuste

Chapter 39: A Practical Guide: Measurement of Free Ca²⁺ Concentration in the Lumen of Neuronal Endoplasmic Reticulum
N. Solovyova and A. Verkhratsky

Chapter 40: Imaging Intracellular Calcium–concentration Microdomains at a Chemical Synapse
R. Llinás and M. Sugimori

Chapter 41: Monitoring Intramitochondrial Calcium with Rhod–2
M. Hoth and R.S. Lewis

Chapter 42: Generation of Controlled Calcium Oscillations in Nonexcitable Cells
R.E. Dolmetsch and R.S. Lewis

Chapter 43: A Practical Guide: High–Speed Imaging of Calcium Waves in Neurons in Brain Slices
W.N. Ross, T. Nakamura, S. Watanabe, M.E. Larkum, and N. Lasser–Ross

Chapter 44: A Practical Guide: Imaging Action Potentials with Calcium Indicators
J.N. MacLean and R. Yuste

Chapter 45: Imaging Calcium Transients in Developing Xenopus Spinal Neurons
N.C. Spitzer, L.N. Borodinsky, and C.M. Root

SECTION 4: PHOTOACTIVATION

Chapter 46: Basics of Photoactivation
G.C.R. Ellis–Davies

Chapter 47: Two–Photon Uncaging Microscopy
H. Kasai, M. Matsuzaki, and G.C.R. Ellis–Davies

Chapter 48: A Practical Guide: Chemical Two–Photon Uncaging
D.L. Pettit and G.J. Augustine

Chapter 49: A Practical Guide: Uncaging with Visible Light, Inorganic Caged Compounds
L. Zayat, L. Baraldo, and R. Etchenique

Chapter 50: A Practical Guide: Infrared–guided Laser Stimulation of Neurons in Brain Slices
H.–U. Dodt, M. Eder, A. Schierloh, and W. Zieglgänsberger

Chapter 51: Uncaging Calcium in Neurons
K.R. Delaney and V. Shahrezaei

Chapter 52: A Practical Guide: Building a Simple Uncaging System
K. Kandler, G. Kim, and B. Schmidt

Chapter 53: Ca²⁺ Uncaging in Nerve Terminals
R. Schneggenburger

Chapter 54: Direct Multiphoton Stimulation of Neurons and Spines
H. Hirase, V. Nikolenko, and R. Yuste

SECTION 5: ADVANCED IMAGING AND SPECIAL APPLICATIONS

Chapter 55: A Practical Guide: Imaging Microglia in Live Brain Slices and Slice Cultures
D. Kurpius and M.E. Dailey

Chapter 56: Single and Multiphoton Fluorescence Recovery after Photobleaching
E. Brown, A. Majewska, and R.K. Jain

Chapter 57: All–Optical, In Situ Histology of Neuronal Tissue with Ultrashort Laser Pulses
P.S. Tsai, B. Friedman, C.B. Schaffer, J.A. Squier, and D. Kleinfeld

Chapter 58: Imaging with Voltage–sensitive Dyes: Spike Signals, Population Signals, and Retrograde Transport
E.K. Kosmidis, L.B. Cohen, C.X. Falk, J.–Y. Wu, and B.J. Baker

Chapter 59: Dendritic Voltage Imaging
M. Djurisic, S. Antic, and D. Zecevic

Chapter 60: Second Harmonic Imaging of Membrane Potential
A.C. Millard, A Lewis, and L.M. Loew

Chapter 61: Imaging Synaptic Vesicle Dynamics with Styryl Dyes
S.O. Rizzoli, U. Becherer, J. Angleson, and W.J. Betz

Chapter 62: A Practical Guide: Imaging FM Dyes in Brain Slices
A.R. Kay

Chapter 63: A Practical Guide: Imaging Zinc in Brain Slices
A.R. Kay

Chapter 64: A Practical Guide: Imaging Exocytosis with Total Internal Reflection Microscopy
D. Zenisek and D. Perrais

Chapter 65: A Practical Guide: Measuring Light–scattering Changes Associated with Secretion from Nerve Terminals
B.M. Salzberg, M. Muschol, and A.L. Obaid

Chapter 66: Imaging with Quantum Dots
J.K. Jaiswal, E.R. Goldman, H. Mattoussi, and S.M. Simon

Chapter 67: Imaging Single Receptors with Quantum Dots
S. Lévi, M. Dahan, and A. Triller

Chapter 68: A Practical Guide: Tracking Receptors by Imaging Single Molecules
L. Cognet, B. Lounis, and D. Choquet

Chapter 69: A Practical Guide: Imaging Sodium in Dendrites
W. Ross, J.C. Callaway, and N. Lasser–Ross

Chapter 70: A Practical Guide: Two–Photon Sodium Imaging in Dendritic Spines
C.R. Rose

Chapter 71: Two–Photon Imaging of Chloride
O. Garaschuk and A. Konnerth

Chapter 72: A Practical Guide: Interferometric Detection of Action Potentials
A. LaPorta and D. Kleinfeld

Chapter 73: A Practical Guide: Intrinsic Optical Signal Imaging in Brain Slices
B.A. MacVicar and S.J. Mulligan

SECTION 6: GENETICALLY ENGINEERED FLUORESCENT PROBES

Chapter 74: Indicators Based on Fluorescence Resonance Energy Transfer
R.Y. Tsien

Chapter 75: Cellular Imaging of Bioluminescence
J.D. Plautz and S.A. Kay

Chapter 76: Introduction of Green Fluorescent Protein into Hippocampal Neurons through Viral Infection
R. Malinow, Y. Hayashi, M. Maletic–Savatic, S.H. Zaman, J.–C. Poncer, S.–H. Shi, J.A. Esteban, P. Osten, and K. Seidenman

Chapter 77: Green Fluorescent Proteins for Measuring Voltage
M.S. Siegel and E.Y. Isacoff

Chapter 78: Genetic Probes for Calcium Dynamics
A. Miyawaki, T. Nagai, and H. Mizuno

Chapter 79: A Practical Guide: Targeted Recombinant Aequorins
T. Pozzan and R. Rizzuto

Chapter 80: A Practical Guide: Imaging Synaptic Inhibition with Clomeleon, a Genetically Encoded Chloride Indicator
K. Berglund, R.L. Dunbar, P. Lee, G. Feng, and G.J. Augustine

Chapter 81: A Practical Guide: Synapto–pHluorins—Genetically Encoded Reporters of Synaptic Transmission
G. Miesenböck

Chapter 82: Imaging Gene Expression in Live Cells and Tissues
R.E. Dolmetsch, N. Gomez–Ospina, E. Green, and E.A. Nigh

Chapter 83: Imaging Olfactory Activity in Drosophila CNS with a Calcium–sensitive Green Fluorescent Protein
A.M. Wong, J. Flores, and J.W. Wang

Chapter 84: Tracking Molecules in Intact Zebrafish
R. Armisen, M.R. Gleason, J.R. Fetcho, and G. Mandel

SECTION 7: IN VIVO IMAGING

Chapter 85: Long–Term, High–Resolution Imaging of Neurons in the Neocortex In Vivo
A.J.G.D. Holtmaat, L. Wilbrecht, A. Karpova, C. Portera–Cailliau, B. Burbach, J.T. Trachtenberg, and K. Svoboda

Chapter 86: A Two–Photon Fiberscope for Imaging in Freely Moving Animals
F. Helmchen and W. Denk

Chapter 87: A Practical Guide: In Vivo Two–Photon Calcium Imaging Using Multicell Bolus Loading
O. Garaschuk and A. Konnerth

Chapter 88: A Practical Guide: In Vivo Calcium Imaging in the Fly Visual System
A. Borst, W. Denk, and J. Haag

Chapter 89: Intrinsic Signal Imaging in the Neocortex: Implications for Hemodynamic–based Functional Imaging
A. Grinvald, D. Sharon, H. Slovin, and I. Vanzetta

Chapter 90: Voltage–sensitive Dye Imaging of Neocortical Activity
A. Grinvald, D. Sharon, A. Sterkin, H. Slovin, and R. Hildesheim

Chapter 91: A Practical Guide: Whole–Cell Recording and Voltage–sensitive Dye Imaging In Vivo
C. Petersen

Chapter 92: A Practical Guide: In Vivo Imaging of Tumors
E. Brown, L.L. Munn, D. Fukumura, and R.K. Jain

Chapter 93: Two–Photon Imaging of Cortical Microcirculation
D. Kleinfeld and W. Denk

Chapter 94: Imaging Neuronal Activity with Calcium Indicators in Larval Zebrafish
J.R. Fetcho

SECTION 8: PRINCIPLES AND INSTRUMENTATION

Chapter 95: Microscopy and Microscope Optical Systems
F. Lanni and H.E. Keller

Chapter 96: Practical Limits to Resolution in Fluorescence Light Microscopy
E.H.K. Stelzer

Chapter 97: Lasers for Multiphoton Microscopy
F.W. Wise

Chapter 98: Acousto–optic Tunable Filters for Microscopy
E.S. Wachman

Chapter 99: A Practical Guide: Arc Lamps and Monochromators for Fluorescence Microscopy
R. Uhl

Chapter 100: The Use of Liquid–Crystal Tunable Filters for Fluorescence Imaging
K.R. Spring

Chapter 101: Fluorescence Grating Imager Systems for Optical–sectioning Microscopy
F. Lanni

Chapter 102: Analysis of Dynamic Optical Imaging Data
B. Pesaran, A.T. Sornborger, N. Nishimura, D. Kleinfeld, and P.P. Mitra

Appendices

1. Electromagnetic Spectrum

2. Microscopy: Lenses, Filters, and Emission/Excitation Spectra

3. Cautions

Index