A novel method for measuring dynamic changes in cell volume.
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.
Department of Biomedical Engineering, University of Virginia Health Sciences Center, Charlottesville, Virginia 22908, USA.
Cristina E
CE
Davis
Joshua J
JJ
Rychak
Bouvard
B
Hosticka
Scott C
SC
Davis
J Edward
JE
John
Amy L
AL
Tucker
Pamela M
PM
Norris
J Randall
JR
Moorman
D000818:Animals; D019556:COS Cells; D002138:Calibration; D048429:Cell Size; D002522:Chlorocebus aethiops; D003584:Cytological Techniques; D007668:Kidney / Q000166:cytology* / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D008954:Models, Biological; D010750:Phosphoproteins / Q000378:metabolism; D014160:Transducers, Pressure
D008565:Membrane Proteins; D010750:Phosphoproteins; C069141:phospholemman