Chapter 13 Perfusion Chambers for High-Resolution Video Light Microscopic Studies of Vertebrate Cell Monolayers: Some Considerations and a Design

Abstract

Publisher Summary This chapter defines a perfusion chamber as a vessel, mounted on the stage of a light microscope that enables a living specimen to be followed continuously by high-resolution light microscopy (LM) while also allowing the fluid bathing the specimen to be rapidly exchanged during the observation period. Perfusion chambers are employed in a variety of biological and biomedical studies of cells and tissues. As they allow the media bathing the specimen to be removed and replenished, they are widely used to maintain specimen viability during long-term investigations, to study the effects of environmental changes, and to periodically sample for the production of cellular products. Perfusion chambers are also employed to deliver tracers for uptake studies and to assay the effects of drugs and other biologically active components on cell/tissue structure and function. The chapter discusses the topic in two-fold manner: (1) the first is to identify some of the variables that must be considered when constructing or choosing a perfusion chamber for high-resolution video-enhanced LM studies. (2) The second is to describe a rugged and simple laminar-flow chamber used in our laboratory for short-term (12-day) studies of vertebrate cell monolayers.