DETERMINATION OF VOLUME AND WATER PERMEABILITY OF PLATED CELLS FROM MEASUREMENTS OF LIGHT-SCATTERING

Citation
J. Fischbarg et al., DETERMINATION OF VOLUME AND WATER PERMEABILITY OF PLATED CELLS FROM MEASUREMENTS OF LIGHT-SCATTERING, The American journal of physiology, 265(5), 1993, pp. 30001412-30001423
Citations number
37
Categorie Soggetti
Physiology
ISSN journal
00029513
Volume
265
Issue
5
Year of publication
1993
Part
1
Pages
30001412 - 30001423
Database
ISI
SICI code
0002-9513(1993)265:5<30001412:DOVAWP>2.0.ZU;2-P
Abstract
Measurements of cell membrane water osmotic permeabilities can be inac curate because of the technical difficulties inherent to cell volume m easurements and because of the presence of an unstirred water layer in contact with the cells. We detail here a method we have developed to quantify transient changes in cell volumes from the intensity of light scattered by cells. For this, we theorize how an unstirred layer orig inates in a perfusing chamber, and we calculate values for both cell m embrane water osmotic permeability and unstirred layer thickness from time transient changes in scattered light. We apply a computer algorit hm that finds the best correspondence between experimental data and es timated values. This is done by solving a differential equation govern ing cell volume changes by numerical integration (Runge-Kutta) and ite rating the procedure varying the test values of osmotic permeability a nd unstirred layer thickness until the best fit is achieved. We exempl ify this procedure with experimental results obtained in adherent cult ured cells.