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Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah, Salt Lake City, Utah 84112; and University Laboratory of Physiology, Oxford OX1 3PT, United Kingdom
This study describes the use of a microperfusion system to create rapid, large regional changes in intracellular pH (pHi) within single ventricular myocytes. The spatial distribution of pHi in single myocytes was measured with seminaphthorhodafluor-1 fluorescence using confocal imaging. Changes in pHi were induced by local external application of NH4Cl, CO2, or sodium propionate. Local application was achieved by simultaneously directing two parallel square microstreams, each 275 µm wide, over a single myocyte oriented perpendicular to the direction of flow. One stream contained the control solution, and the other contained a weak acid or base. End-to-end, stable pHi gradients as large as 1 pH unit were readily created with this technique. This result indicates that pH within a single cardiac cell may not always be spatially uniform, particularly when weak acid or base gradients are present, which can occur, for example, in regional myocardial ischemia. The microperfusion method should be useful for studying the effects of localized acidosis on myocyte function, estimating intracellular ion diffusion rates, and, possibly, inducing regional changes in other important intracellular ions.
confocal imaging; intracellular acidosis; ventricular myocytes
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