A novel method for measuring dynamic changes in cell volume

1 Departments of Biomedical Engineering, 3 Internal Medicine (Cardiovascular Division), 4 Molecular Physiology and Biological Physics, and the Cardiovascular Research Center, and 2 Mechanical, Aerospace and Nuclear Engineering, University of Virginia Health Sciences Center, Charlottesville, Virginia 22908 Submitted 14 March 2003 ; accepted in final form 25 October 2003 Many cell types regulate their volume in response to extracellular tonicity changes through a complex series of adaptive mechanisms. Several methods that are presently used to measure cell volume changes include Coulter counters, fluorescent techniques, electronic impedance, and video microscopy. Although these methods are widely used and accepted, there are limitations associated with each technique. This paper describes a new method to measure changes in cell volume based on the principle that fluid flow within a rigid system is well determined. For this study, cos-7 cells were plated to line the inner lumen of a glass capillary and stimulated to swell or shrink by altering the osmolarity of the perfusing solution. The cell capillary was connected in series with a blank reference capillary, and differential pressure changes across each tube were monitored. The advantages of this method include 1 ) ability to continuously monitor changes in volume during rapid solution changes, 2 ) independence from cell morphology, 3 ) presence of physiological conditions with cell surface contacts and cell-cell interactions, 4 ) no phototoxic effects such as those associated with fluorescent methods, and 5 ) ability to report from large populations of cells. With this method, we could detect the previously demonstrated enhanced volume regulation of cells overexpressing the membrane phosphoprotein phospholemman, which has been implicated in osmolyte transport. regulatory volume decrease; cell swelling; phospholemman Address for reprint requests and other correspondence: J. R. Moorman, Box 6012, MR4 Bldg., UVAHSC, Charlottesville, VA 22908 (E-mail: rmoorman{at}virginia.edu ).