Journal of Applied Physiology
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J Appl Physiol 76: 1394-1399, 1994;
8750-7587/94 $5.00
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Journal of Applied Physiology, Vol 76, Issue 3 1394-1399, Copyright © 1994 by American Physiological Society

ARTICLES

Fura 2 determination of [Ca2+]i in isolated perfused heart using R wave-gated electromechanical shutters

D. P. Chen, E. Jimenez, K. Ataka, S. Levitsky and H. Feinberg
Division of Cardiothoracic Surgery, New England Deaconess Hospital, Boston, Massachusetts 02215.

We describe a novel and relatively inexpensive spectrofluorescence system that supplies rapidly alternating wavelengths to either a standard cuvette or an isolated perfused heart. Its use is illustrated by determining changes in cytosolic intracellular Ca2+ concentration ([Ca2+]i) by using the Ca(2+)-sensitive fluorescent dye fura 2 in a rabbit heart preparation. The system uses two precision electromechanical shutters (capable of gating with respect to the electrocardiographic R wave for signal averaging) allowing alternate fura 2 excitation wavelengths (340 and 380 nm) without moving optical components and uses a fiber optic for conducting excitation and collecting epifluorescence. Sample recordings tracing the [Ca2+]i transient in an entire cardiac cycle and in capturing specific isolated regions (diastole and systole) of the cycle are presented. Limitations of this low-cost but easily implemented system are discussed.


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