title: Methods for two-dimensional cell confinement.
abstract: Protocols described in this chapter relate to a method to dynamically confine cells in two dimensions with various microenvironments. It can be used to impose on cells a given height, with an accuracy of less than 100 nm on large surfaces (cm(2)). The method is based on the gentle application of a modified glass coverslip onto a standard cell culture. Depending on the preparation, this confinement slide can impose on the cells a given geometry but also an environment of controlled stiffness, controlled adhesion, or a more complex environment. An advantage is that the method is compatible with most optical microscopy technologies and molecular biology protocols allowing advanced analysis of confined cells. In this chapter, we first explain the principle and issues of using these slides to confine cells in a controlled geometry and describe their fabrication. Finally, we discuss how the nature of the confinement slide can vary and provide an alternative method to confine cells with gels of controlled rigidity.
authors.affiliation: Systems Cell Biology of Cell Division and Cell Polarity, Institut Curie, CNRS, Paris, France.
authors.forename: Maël
authors.initials: M
authors.lastname: Le Berre
authors.affiliation: Systems Cell Biology of Cell Division and Cell Polarity, Institut Curie, CNRS, Paris, France.
authors.forename: Ewa
authors.initials: E
authors.lastname: Zlotek-Zlotkiewicz
authors.affiliation: Systems Cell Biology of Cell Division and Cell Polarity, Institut Curie, CNRS, Paris, France.
authors.forename: Daria
authors.initials: D
authors.lastname: Bonazzi
authors.affiliation: Systems Cell Biology of Cell Division and Cell Polarity, Institut Curie, CNRS, Paris, France.
authors.forename: Franziska
authors.initials: F
authors.lastname: Lautenschlaeger
authors.affiliation: Systems Cell Biology of Cell Division and Cell Polarity, Institut Curie, CNRS, Paris, France.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Piel
mesh_terms: D000180:Acrylic Resins / Q000737:chemistry; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D002465:Cell Movement; D002478:Cells, Cultured; D020378:Confined Spaces; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D006801:Humans; D008853:Microscopy / Q000295:instrumentation / Q000379:methods; D013314:Stress, Mechanical
chemical_list: D000180:Acrylic Resins; D004129:Dimethylpolysiloxanes; C016680:polyacrylamide gels; C013830:baysilon
keywords: Biophysics; Cell adhesion; Cell mechanics; Cell migration; Gel assay; Microenvironment; Microfabrication