Multiphoton Microscopy

370 Pages·2019·27.85 MB·English

by  Espen Hartveit

Neuromethods 148
Espen Hartveit
Editor
Multiphoton
Microscopy
N
euromethods
Series Editor
Wolfgang Walz
University of Saskatchewan
Saskatoon, SK, Canada
For further volumes:
http://www.springer.com/series/7657
Neuromethods publishes cutting-edge methods and protocols in all areas of neuroscience as
well as translational neurological and mental research. Each volume in the series offers tested
laboratory protocols, step-by-step methods for reproducible lab experiments and addresses
methodological controversies and pitfalls in order to aid neuroscientists in experimentation.
Neuromethods focuses on traditional and emerging topics with wide-ranging implications to
brain function, such as electrophysiology, neuroimaging, behavioral analysis, genomics,
neurodegeneration, translational research and clinical trials. Neuromethods provides inves-
tigators and trainees with highly useful compendiums of key strategies and approaches for
successful research in animal and human brain function including translational “bench to
bedside” approaches to mental and neurological diseases.
Multiphoton Microscopy
Edited by
Espen Hartveit
Department of Biomedicine, University of Bergen, Bergen, Norway
Editor
Espen Hartveit
Department of Biomedicine
University of Bergen
Bergen, Norway
ISSN 0893-2336 ISSN 1940-6045 (electronic)
Neuromethods
ISBN 978-1-4939-9701-5 ISBN 978-1-4939-9702-2 (eBook)
https://doi.org/10.1007/978-1-4939-9702-2
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Preface to the Series
Experimental life sciences have two basic foundations: concepts and tools. The Neuromethods
series focuses on the tools and techniques unique to the investigation of the nervous system
and excitable cells. It will not, however, shortchange the concept side of things as care has
been taken to integrate these tools within the context of the concepts and questions under
investigation. In this way, the series is unique in that it not only collects protocols but also
includes theoretical background information and critiques which led to the methods and
their development. Thus it gives the reader a better understanding of the origin of the
techniques and their potential future development. The Neuromethods publishing pro-
gram strikes a balance between recent and exciting developments like those concerning
new animal models of disease, imaging, in vivo methods, and more established techniques,
including, for example, immunocytochemistry and electrophysiological technologies. New
trainees in neurosciences still need a sound footing in these older methods in order to apply
a critical approach to their results.
Under the guidance of its founders, Alan Boulton and Glen Baker, the Neuromethods
series has been a success since its first volume published through Humana Press in 1985.
The series continues to flourish through many changes over the years. It is now published
under the umbrella of Springer Protocols. While methods involving brain research have
changed a lot since the series started, the publishing environment and technology have
changed even more radically. Neuromethods has the distinct layout and style of the Springer
Protocols program, designed specifically for readability and ease of reference in a laboratory
setting.
The careful application of methods is potentially the most important step in the process
of scientific inquiry. In the past, new methodologies led the way in developing new disci-
plines in the biological and medical sciences. For example, physiology emerged out of
anatomy in the nineteenth century by harnessing new methods based on the newly discov-
ered phenomenon of electricity. Nowadays, the relationships between disciplines and meth-
ods are more complex. Methods are now widely shared between disciplines and research
areas. New developments in electronic publishing make it possible for scientists that
encounter new methods to quickly find sources of information electronically. The design of
individual volumes and chapters in this series takes this new access technology into account.
Springer Protocols makes it possible to download single protocols separately. In addition,
Springer makes its print-on-demand technology available globally. A print copy can there-
fore be acquired quickly and for a competitive price anywhere in the world.
Saskatoon, SK, Canada Wolfgang Walz
v
Preface
The ability to visualize live and intact neural tissue at the cellular and subcellular level has
seen incredible advances over the past two decades. One of the core techniques driving this
continued progression and development is multiphoton excitation (MPE) microscopy. It is
remarkable that a clever application of a fundamental principle in physics has provided this
powerful tool to visualize and interrogate complex neural tissue in 3D. For those of us who
stubbornly insist on working with intact neural tissue, either a slice preparation in vitro or
a whole animal in vivo, the ability to look deep into the tissue with detailed subcellular reso-
lution is nothing less than magic. For a young scientist entering the field today, where MPE
microscopy can be found at almost every neuroscience research center, it must be difficult
to imagine the momentous transition into the MPE world of today when fluorescence
imaging of live neural tissue with supreme optical sectioning and minimal phototoxicity was
enabled.
During the early years, establishing a laboratory for MPE microscopy was very costly
and demanded specialized technical skills (or an in-house physicist) to manage the neces-
sary lasers. But this has changed, and the technique, although still on the expensive side for
individual laboratories, has matured and become much more accessible. Nevertheless, even
with a number of turnkey systems offered by commercial companies, many scientists still
decide to build their own microscopes. For readers who would like to try this, the first
chapter offers some invaluable advice.
My primary goal in editing this book was to assemble a collection of chapters covering
different applications of MPE microscopy that would highlight techniques and experimen-
tal strategies that expert researchers have found to be useful and successful in their own
work and laboratories. I have encouraged the authors to include as much detail as possible
when it comes to practical and effective “tips and tricks” that hopefully will benefit both
established researchers and newcomers with little or no previous experience in MPE micros-
copy. For some applications, this has resulted in chapters with a series of step-by-step pro-
cedures. For other applications, it has resulted in detailed discussions and explanations of
creative solutions to difficult problems. In both cases, I hope the chapters will inspire,
guide, and ease the path from novice to master. Essentially, I wanted to produce the book
I would have liked to read myself, more than 10 years ago when I eagerly unpacked and
assembled the “Movable Objective Microscope” (also known as the “Denk scope”) from
Sutter Instrument. Whereas the research and applications enabled by MPE microscopy are
not limited to neuroscience, the list of important discoveries in this field is already long and
continues to increase.
In this book, the reader will find chapters that together cover the fundamentals of MPE
microscopy as applied to both in vitro and in vivo experimental systems. Scientific advances
often involve combining different techniques, and several chapters provide information on
how to combine MPE microscopy with targeted electrophysiological recording and with
synthetic and genetically encoded sensors for signaling molecules, calcium, and transmem-
brane voltage. Pulsed laser light is useful not only for imaging but also for uncaging neuro-
active substances, and this is covered in several chapters that illustrate the application of this
vii
viii Preface
technique for investigating neural signaling and plasticity. Additional chapters provide valu-
able information for using MPE microscopy to investigate cellular and large-scale neural
morphology, signaling in astrocytes, and multi-site signaling with subcellular resolution in
neural networks of behaving animals. Finally, no book on MPE microscopy is complete
without a chapter detailing how this imaging technique was first used, and continues to be
used, to study the retina, the body’s own light sensor and image processor.
I would like to thank the series editor, Wolfgang Walz, for his patience, guidance, and
encouragement during this project. Last, but not least, I would also like to take this oppor-
tunity to express my sincere gratitude to each of the authors for their efforts in providing
their valuable contributions to this volume which I hope will enable many more to enjoy
the benefits of MPE microscopy.
Bergen, Norway Espen Hartveit
Contents
Preface to the Series ................................................ v
Preface ......................................................... vii
Contributors ..................................................... xi
1 Building a Two-Photon Microscope Is Easy .......................... 1
Spencer LaVere Smith
2 Using Multiphoton Imaging for Targeted Electrophysiological
Recording and Live Cell Imaging of Fluorescently Labeled
Neurons from Isolated Retinas .................................... 17
Qiang Chen and Wei Wei
3 Two-Photon Neurotransmitter Uncaging for the Study of Dendritic
Integration. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
Alexandra Tran-Van-Minh, Nelson Rebola, Andreas Hoehne,
and David A. DiGregorio
4 Two-Photon Glutamate Uncaging to Study Structural
and Functional Plasticity of Dendritic Spines ......................... 65
Ivar S. Stein, Travis C. Hill, Won Chan Oh, Laxmi K. Parajuli,
and Karen Zito
5 Two-Photon Fluorescence Imaging of Visually Evoked Glutamate
Release Using iGluSnFR in the Mouse Visual System ................... 87
Bart G. Borghuis
6 Imaging Neuronal Signal Transduction Using
Multiphoton FRET-FLIM ....................................... 111
Paul R. Evans, Long Yan, and Ryohei Yasuda
7 Combining Multiphoton Excitation Microscopy with Fast
Microiontophoresis to Investigate Neuronal Signaling .................. 131
Espen Hartveit and Margaret Lin Veruki
8 Multiphoton Excitation Microscopy for the Reconstruction
and Analysis of Single Neuron Morphology .......................... 161
Espen Hartveit, Bas-Jan Zandt, and Margaret Lin Veruki
9 Serial Multiphoton Tomography and Analysis of Volumetric
Images of the Mouse Brain ...................................... 195
Denise M. O. Ramirez, Apoorva D. Ajay, Mark P. Goldberg,
and Julian P. Meeks
10 Studying a Light Sensor with Light: Multiphoton Imaging in the Retina .... 225
Thomas Euler, Katrin Franke, and Tom Baden
11 Multiphoton Ca2+ Imaging of Astrocytes with Genetically Encoded
Indicators Delivered by a Viral Approach ............................ 251
Rune Enger, Rolf Sprengel, Erlend A. Nagelhus, and Wannan Tang
ix
x Contents
12 Two-Photon Imaging of Dendritic Calcium Dynamics In Vivo ............ 279
Lucy M. Palmer
13 Voltage Imaging with ANNINE Dyes and Two-Photon Microscopy ....... 297
Christopher J. Roome and Bernd Kuhn
14 High-Speed, Random-Access Multiphoton Microscopy for Monitoring
Synaptic and Neuronal Activity in 3D in Behaving Animals .............. 335
Mate Marosi, Gergely Szalay, Gergely Katona, and Balázs Rózsa
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 363