Laser Scanning Microscopy and Quantitative Image Analysis of Neuronal Tissue

321 Pages·2014·11.682 MB·English

by  Lidia Bakota, Roland Brandt (eds.)

Neuromethods 87
Lidia Bakota
Roland Brandt Editors
Laser Scanning
Microscopy
and Quantitative
Image Analysis
of Neuronal Tissue
N
EUROMETHODS
Series Editor
Wolfgang Walz
University of Saskatchewan
Saskatoon, SK, Canada
For further volumes:
h ttp://www.springer.com/series/7657

Laser Scanning Microscopy
and Quantitative Image Analysis
of Neuronal Tissue
Edited by
Lidia Bakota
Department of Neurobiology, University of Osnabrück, Osnabrück, Germany
Roland Brandt
Department of Neurobiology, University of Osnabrück, Osnabrück, Germany
Editors
Lidia B akota Roland Brandt
Department of Neurobiology Department of Neurobiology
University of Osnabrück University of Osnabrück
Osnabrück, Germany Osnabrück, Germany
ISSN 0893-2336 ISSN 1940-6045 (electronic)
ISBN 978-1-4939-0380-1 ISBN 978-1-4939-0381-8 (eBook)
DOI 10.1007/978-1-4939-0381-8
Springer New York Heidelberg Dordrecht London
Library of Congress Control Number: 2014931752
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Preface to t he Series
Under the guidance of its founders Alan Boulton and Glen Baker, the Neuromethods series
by Humana Press has been very successful since the fi rst volume appeared in 1985. In about
17 years, 37 volumes have been published. In 2006, Springer Science + Business Media
made a renewed commitment to this series. The new program will focus on methods that
are either unique to the nervous system and excitable cells or which need special consider-
ation to be applied to the neurosciences. The program will strike a balance between recent
and exciting developments like those concerning new animal models of disease, imaging, in
vivo methods, and more established techniques. These include 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. The careful
application of methods is probably the most important step in the process of scientifi c
inquiry. In the past, new methodologies led the way in developing new disciplines 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 discovered phenome-
non of electricity. Nowadays, the relationships between disciplines and methods are more
complex. Methods are now widely shared between disciplines and research areas. New
developments in electronic publishing also make it possible for scientists to download chap-
ters or protocols selectively within a very short time of encountering them. This new
approach has been taken into account in the design of individual volumes and chapters in
this series.
Saskatoon, S K , C anada Wolfgang Walz
v

Prefa ce
Laser scanning microscopy (LSM) has become an established and broadly used method to
obtain optical images with depth selectivity. LSM includes confocal laser scanning micros-
copy (cLSM) as its most common and commercially mature application, but also multipho-
ton microscopy gets increasingly used because it allows for obtaining images from living
specimens with minimal phototoxicity and increased tissue penetration. The key features of
both the techniques are that they permit optical sectioning, that data are obtained by scan-
ning in a point-by-point fashion, and that images are then reconstructed by electronic
means, which generates a digital data set that allows three-dimensional reconstructions and
further analysis of complex objects.
Many novel advances in LSM are due to increased digital data processing power and
mathematical analysis of the data sets, which allows users, for example, to perform 4D
recordings, i.e., to follow 3D reconstructions over time in order to determine the dynamics
of biological processes and scrutinize morphogenetic mechanisms. In addition, quantitative
colocalization and connectivity analyses have become possible. Such applications are in
particular useful for research in neuroscience, which deals with one of the most complex
tissue, different cell types with extensive morphological differentiation and signifi cant struc-
tural plasticity during nervous system development, maintenance, and aging.
The aim of this book is to introduce applications and quantitative image analysis using
data sets obtained from confocal and multiphoton laser-scanning microscopy of neuronal
tissue. The idea is to introduce key biological questions, to provide detailed information
how to acquire data by laser-scanning microscopy, and to examine how to use the often
huge digital data set in an effi cient manner to extract maximum of information. Thus the
book not only provides a compilation of diverse protocols but also aims to bring together
biological bench work, laser scanning microscopy, and mathematical, computer-assisted
data analysis to grasp novel insights of form, dynamics, and interactions of microscopy-
sized biological objects. We are confi dent that the scope of this book goes much beyond
current protocols compilations that introduce selected lab and microscopy techniques. We
hope that the book will stimulate the reader to make effi cient use of the application of laser
scanning microscopy for his or her own research question. To achieve this goal, we have
brought together contributions from research institutions all over the world covering pio-
neering applications in laser scanning microscopy and quantitative image analysis, and pro-
viding information about the power and limitations of this quickly developing fi eld at the
interface between biology, physics, informatics, and mathematics.
Osnabrück, G ermany L idia Bakota
Roland Brandt
vii

Contents
Preface to the Series. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix
1 Translation, Touch, and Overlap in Multi-fluorescence Confocal
Laser Scanning Microscopy to Quantitate Synaptic Connectivity. . . . . . . . . . . 1
Floris G. Wouterlood and Jeroen A.M. Beliën
2 Surgical Procedures to Study Microglial Motility in the Brain
and in the Spinal Cord by In Vivo Two-Photon
Laser- Scanning Microscopy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
Alexander Cupido, Bogdan Catalin, Heinz Steffens,
and Frank Kirchhoff
3 Analysis of Brain Projection Systems Using Third-G eneration
Neuroanatomical Tracers and Multiple Fluorescence
Laser Scanning Microscopy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
Floris G. Wouterlood
4 Combining Multichannel Confocal Laser Scanning Microscopy
with Serial Section Reconstruction to Analyze Large Tissue Volumes
at Cellular Resolution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
Federico Luzzati
5 Modeling Excitotoxic Ischemic Brain Injury of Cerebellar Purkinje
Neurons by Intravital and In Vitro Multi-photon Laser
Scanning Microscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105
Amanda J. Craig, Gary D. Housley, and Thomas Fath
6 Analysis of Morphology and Structural Remodeling of Astrocytes . . . . . . . . . . 129
Tatjana C. Jakobs
7 Quantitative Analysis of Axonal Outgrowth in Mice . . . . . . . . . . . . . . . . . . . . 145
Rosa-Eva Huettl and Andrea B. Huber
8 Zebrafish Brain Development Monitored by Long-Term
In Vivo Microscopy: A Comparison Between Laser
Scanning Confocal and 2-Photon Microscopy. . . . . . . . . . . . . . . . . . . . . . . . . 163
Nicolas Dross, Carlo Antonio Beretta, Peter Bankhead,
Matthias Carl, and Ulrike Engel
9 Analysis of Actin Turnover and Spine Dynamics
in Hippocampal Slice Cultures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189
Kristin Michaelsen-Preusse, Yves Kellner, Martin Korte,
and Marta Zagrebelsky
ix