Chapter 3
Micromechanical Control of Cell-Cell Interactions

3.1 Introduction 3.1.1 Cell-cell interactions 3.1.2 Conventional co-cultivation models 3.1.3 Micromechanical reconfigurable culture 3.1.4 Application examples 3.1.4.1 Cell patterning 3.1.4.2 Decoupling of contact-mediated and soluble signals 3.1.4.3 Range of soluble signaling 3.1.4.4 Temporal control of cell-cell signaling 3.1.4.5 Deconvolution of overlapping cell processes 3.1.4.6 Separation of cell populations 3.1.4.7 Population-specific stimulation and interrogation 3.1.4.8 Investigation of reciprocal signaling and cell-cell crosstalk 3.2 Experimental Design 3.2.1 Experimental variables 3.2.2 Readout 3.3 Materials 3.3.1 Reagents/Supplies 3.3.2 Facilities/Equipment 3.4 Methods 3.4.1 Device handling and actuation 3.4.2 Preparing devices for cell culture 3.4.3 Cell seeding 3.4.4 Assay preparation 3.5 Discussion 3.6 Troubleshooting Table 3.7 Summary Points

Chapter 4
Mechanotransduction and the Study of Cellular Forces

4.1		Introduction
4.1.1		Cellular Forces: Functions and Underlying Mechanisms
4.1.2		Techniques for Studying Traction Forces
4.2		Materials
4.2.1		Reagents/Supplies
4.2.2		Facilities/Equipment/Software
4.3		Methods
4.3.1		Microfabrication of Micropost Arrays
4.3.1.1		Rationale for Different Master Fabrication Processes
4.3.1.2		Reticle Design
4.3.1.3		Projection Photolithography of Micropost Arrays
4.3.1.4		Deep Reactive Ion Etching of Silicon Microposts
4.3.1.5		Device Packaging
4.3.1.6		Soft Lithography of Micropost Array Substrates
4.3.1.7		Characterization of Micropost Spring Constant
4.3.2		Analysis of Traction Forces with Micropost Arrays
4.3.2.1		Substrate Preparation
4.3.2.2		Staining and Microscopy of Micropost Arrays
4.3.2.3		Image Analysis of Micropost Deflections
4.4		Discussion
4.4.1		Applications and Enhancements of the Micropost Arrays
4.4.1.1		Micropatterning Single Cells and Multicellular Aggregates
4.4.1.2		Magnetic Actuation of Microposts to Investigate Localized Force Application
4.4.2		Potential Pitfalls of Micropost Arrays
4.4.2.1		Accuracy of the Slender Beam Approximation
4.4.2.2		Scaling Down Micropost Array Geometry
4.4.3		Biological Insights from Using Micropost Arrays
4.4.4		Future Innovations for Studying Cellular Forces
4.5		Troubleshooting Table
4.6		Summary Points

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