2014 Volume 2014 Issue 24 Pages 217-221
Cells of the body are exposed to anisosmotic environments under various physiological and pathological conditions, which cause mobilization of water and changes in cellular volume and morphology; these changes provide an indication of the cellular environment changes and predict the subsequent cellular damage. We have constructed confocal microscopy-based methods to analyze osmotic stimuli-induced changes in cell dimension and morphology using a membrane-bound fluorescent probe, PKH67. Low osmolarity stimulation of the renal tube derived MDCK cells resulted in an initial increase in their cell volume, which was followed by a subsequent decrease towards initial value. Transient exposure to high osmolarity formamide solution of mouse ventricular cardiomyocytes resulted in complete loss of their transverse tubules (T-tubules). Confocal microscopy in combination with a membrane-intercalating dye was shown to be simple, accurate and free of cellular damage, and would be useful for further studies on the regulation of cellular volume and morphology.
