Microfluidic and Compartmentalized Platforms for Neurobiological Research

249 Pages·2015·10.094 MB·English

by  Emilia Biffi (eds.)

Neuromethods 103
Emilia Biffi Editor
Microfl uidic and
Compartmentalized
Platforms for
Neurobiological
Research
N
EUROMETHODS
Series Editor
Wolfgang Walz
University of Saskatchewan
Saskatoon, SK, Canada
For further volumes:
h ttp://www.springer.com/series/7657

Microfluidic and
Compartmentalized Platforms
for Neurobiological Research
Edited by
Emilia Biffi
Bioengineering Laboratory, Scientific Institute IRCCS E. Medea, Bosisio Parini (LC), Italy
Department of Electronics, Information and Bioengineering, Politecnico di Milano,
Milan, Italy
Editor
Emilia B iffi
Bioengineering Laboratory
Scientific Institute IRCCS E. Medea
Bosisio Parini (LC), Italy
Department of Electronics
Information and Bioengineering
Politecnico di Milano, Milan, Italy
ISSN 0893-2336 ISSN 1940-6045 (electronic)
Neuromethods
ISBN 978-1-4939-2509-4 ISBN 978-1-4939-2510-0 (eBook)
DOI 10.1007/978-1-4939-2510-0
Library of Congress Control Number: 2015932873
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Series P reface
Experimental life sciences have two basic foundations: concepts and tools. The N euromethods
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 N euromethods publishing program
strikes a balance between recent and exciting developments like those concerning new ani-
mal models of disease, imaging, in vivo methods, and more established techniques, includ-
ing, 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 N euromethods
series has been a success since its fi rst volume published through Humana Press in 1985. The
series continues to fl ourish 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 specifi cally 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 scientifi c 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 fi nd 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.
Wolfgang W alz
v

Prefa ce
Microfl uidics is a technology which features the manipulation of small amounts of fl uids in
channels with dimensions of tens to hundreds of micrometers. Microfl uidics takes advantage
of both soft lithography and poly(dimethylsiloxane) (PDMS), a silicon-based elastomeric
material which is cheap, easy to mold and with good optical properties, as well as nontoxic to
cells and gas permeable, offering a suitable solution for cell and tissue culture experiments.
The design of microfl uidic devices as platforms with different compartments had been
inspired by the so-called Campenot chambers, developed to compartmentalize axons and
cell bodies. Microfabricated versions of these devices have already been used in a wide range
of biological applications thanks to their low consumption of samples and reagents, the
ability to precisely control parameters within the cellular microenvironment, and the capa-
bility to perform highly concurrent and reproducible analyses. Thanks to their scale, com-
patible with neuron size, compartmentalized microfl uidic devices are nowadays a crucial
tool in the fi eld of neuroscience. They indeed provide a powerful platform for the manipu-
lation of subpopulation of neuronal cells and the study of complex system dynamics associ-
ated to the connectivity between two or more adjacent regions of the brain. Thanks to the
feature of fl uidically separated compartments, these devices are also suitable for local cell
stimulation, for the creation of dynamic concentration gradients, or for high-content phar-
macological screening.
In this volume, the main cutting-edge techniques to design and fabricate compartmen-
talized microfl uidic devices are described in P art I . In Chap. 1 , a microfl uidic cell coculture
platform that uses pneumatically or hydraulically controlled valves to reversibly separate cell
populations is described. Chapter 2 illustrates the procedures to fabricate microfl uidic
devices reversibly sealed to different fl at substrates through magnetic forces, which are a
suitable approach for long-term cultures of neurons due to their reliable hydraulic tight-
ness. In Chap. 3 , microfl uidic circuits for arraying neurons with single cell precision,
enabling high-throughput experimentation, are presented as well as approaches to couple
these minimalistic cocultures to open access reservoirs for electrophysiology recordings.
Part II is dedicated to the topic of axon guidance and manipulation. Chapter 4 pro-
vides details of a microfl uidic chip with a modular design for highly defi ned isolation of
axons, asymmetric genetic manipulation, and whole-cell patch-clamp recording. In Chap.
5 a unique laser cell-micropatterning system for the creation of a compartmentalized, axon-
isolating, polarized neuron-growth platform at the single-cell level is described. Chapter 6
explains how Campenot cultures and microfl uidics chambers can be coupled and used
together for biochemical analysis and for high resolution imaging of Dorsal Root Ganglia
(DRG) neuronal cell bodies and their extensive axons.
In Part III , compartmentalized devices for synapse manipulation are described. Chapter
7 illustrates an easy and inexpensive technique based on microfl uidics that provides a high
degree of control in positioning and guiding cells, thereby enabling the laying down of
desired cellular networks and facilitating the study of synaptic connections. In Chap. 8 , the
use of three-compartment microfl uidic devices to model in vitro activity-dependent synap-
tic plasticity with dual inputs is detailed and the synaptic competition model is presented.
vii
viii Preface
Part IV is dedicated to the study of how different cell populations interact in either
physiological or pathological condition. Chapter 9 describes a six-compartment neuron-
glia coculture microsystem platform, where interactions between the axon and glia can be
studied in isolation. In Chap. 1 0 , compartmentalized microfl uidic devices to study periph-
eral neuro-osteogenic interactions are discussed, and qualitative and quantitative analyses
for two- or three-dimensions cocultures are also presented. Chapter 11 provides details of
a compartmented in vitro model of the lower motor neuron-neuromuscular junction cir-
cuit, incorporating primary spinal motor neurons, supporting glia and skeletal muscle, and
spatially mimicking the unique anatomical and cellular interactions of this circuit.
In Part V , compartmentalized devices for pharmacological research and drug discovery
are described. Chapter 1 2 details a powerful tool based on microfl uidics used to mimic the
key pathological hallmarks of Alzheimer’s disease and observe the long-term disease spread-
ing at the microscale. In Chap. 1 3 a device for long-term growth of twin neuronal networks
and for their controlled biochemical stimulation and electrophysiological recording is
described. Finally, in Chap. 14 the protocol and methodological considerations for devel-
oping synapse microarrays enabling ultrasensitive, high-throughput and quantitative
screening of small molecules involved in synaptogenesis are illustrated.
The authors and I believe you will fi nd this volume invaluable in gaining an under-
standing of the practical skills needed to fabricate and use microfl uidics and compartmen-
talized platforms with cell cultures as well as the strengths of these exciting devices and their
precious contribution in the fi eld of neuroscience.
Milan, Italy E milia Biffi
Contents
Series Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi
PART I DESIGNS AND METHODS
1 The Fabrication of Microfluidic Platforms with Pneumatically/Hydraulically
Controlled PDMS Valves and Their Use in Neurobiological Research. . . . . . . 3
Bryson M . Brewer , D onna J . W ebb , and Deyu L i
2 A Reliable Reversible Bonding Method for Perfused Microfluidic Devices. . . . 2 5
Paola Occhetta , Emilia B iffi , and Marco Rasponi
3 Bridging Two Cultures: Minimalistic Networks Prepared by Microfluidic
Arraying, and Open Access Compartments for Electrophysiology . . . . . . . . . . 3 9
Jonathan West , N goc-Duy D inh , Heike H ardelauf , Y a-Yu Chiang ,
Tracey A . N ewman , Mariana V argas-Caballero , A yodeji A. Asuni ,
Katrin D einhardt , and Martin Arundell
PART II AXONAL GUIDANCE AND MANIPULATION
4 Asymmetric Genetic Manipulation and Patch Clamp Recording
of Neurons in a Microfluidic Chip. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
Prasanna S akha , Cecilia B runello , Joonas H eikkinen ,
Ville Jokinen , and Henri J. Huttunen
5 D evelopment of a Compartmentalized Biochip for Axonal Isolation
and Neuronal-Circuit Formation at the Single-Cell Level . . . . . . . . . . . . . . . . 8 3
Ting Huang , Russell K . Pirlo , Wan Q in , Y ongliang L in , L ina W ei ,
Lucas Schmidt , N ick E rdman , T ingfei Xi , Mauris N . D eSilva ,
and Bruce Z. Gao
6 Campenot Cultures and Microfluidics Provide Complementary
Platforms for Spatial Study of Dorsal Root Ganglia Neurons. . . . . . . . . . . . . . 105
Sara J. Fenstermacher , M aria F . P azyra-Murphy , and Rosalind A . S egal
PART III MANIPULATION OF SYNAPSES
7 Development of Microfluidic Devices for the Manipulation
of Neuronal Synapses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 27
Anika J ain and M artha U . Gillette
8 U se of a 3-Compartment Microfluidic Device to Study Activity
Dependent Synaptic Competition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
Ainsley Coquinco and Max C ynader
ix