E-Book Overview
The second edition of Adhesion Protein Protocols combines traditional techniques with cutting-edge and novel techniques that can be adapted easily to different molecules and cell types. The topics discussed in this updated second edition include novel techniques for studying cell-cell adhesion, neutrophil chemotaxis, in vitro assays used to study leukocyte migration through monolayers of cultured endothelial cells, and novel techniques to purify pseudopodia from migratory cells. This book also discusses the study of cell-matrix interaction, RNA interference, fluorescence recovery after photobleaching, actin purification, the application of microarray techniques, and the role of adhesion proteins in the study of proteomics.
The protocols discussed in this volume are suitable for both novice and expert scientists, who will gain further insight into the complex and incompletely understood processes involved in cellular adhesion.
E-Book Content
Bowtie-Shaped Microwells 1 1 Manipulation of Cell–Cell Adhesion Using Bowtie-Shaped Microwells Celeste M. Nelson, Wendy F. Liu, and Christopher S. Chen Summary Traditional methods to study cell–cell adhesion have been limited by their inability to manipulate cell–cell interactions without simultaneously affecting other microenvironmental factors. Here we describe a novel method that enables the culture of cells with precise simultaneous control of both cell–cell and cell–substratum adhesion. Using microfabricated stamps of poly(dimethylsiloxane), we construct bowtie-shaped agarose microwells into which cells can be cultured. The degree to which cells spread is controlled by the size of the microwell; cell–cell contacts form between neighboring cells within the microwell. This chapter describes the details of stamp fabrication, a