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Single-Detector Simultaneous Optical Mapping of V m and [Ca2+]i in Cardiac Monolayers

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Abstract

Simultaneous mapping of transmembrane voltage (V m) and intracellular Ca2+ concentration (Cai) has been used for studies of normal and abnormal impulse propagation in cardiac tissues. Existing dual mapping systems typically utilize one excitation and two emission bandwidths, requiring two photodetectors with precise pixel registration. In this study we describe a novel, single-detector mapping system that utilizes two excitation and one emission band for the simultaneous recording of action potentials and calcium transients in monolayers of neonatal rat cardiomyocytes. Cells stained with the Ca2+-sensitive dye X-Rhod-1 and the voltage-sensitive dye Di-4-ANEPPS were illuminated by a programmable, multicolor LED matrix. Blue and green LED pulses were flashed 180° out of phase at a rate of 488.3 Hz using a custom-built dual bandpass excitation filter that transmitted blue (482 ± 6 nm) and green (577 ± 31 nm) light. A long-pass emission filter (>605 nm) and a 504-channel photodiode array were used to record combined signals from cardiomyocytes. Green excitation yielded Cai transients without significant crosstalk from V m. Crosstalk present in V m signals obtained with blue excitation was removed by subtracting an appropriately scaled version of the Cai transient. This method was applied to study delay between onsets of action potentials and Cai transients in anisotropic cardiac monolayers.

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Acknowledgments

This work was supported in part by American Heart Association predoctoral fellowships to Nima Badie (No. 0715178U) and Luke McSpadden (No. 0715288U), and NIH grant R01HL093711.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Duke University, Durham, NC, 27708, USA

    James A. Scull, Luke C. McSpadden, Herman D. Himel IV & Nima Badie

  2. Department of Biomedical Engineering, Faculty of Cardiology, Duke University, Durham, NC, 27708, USA

    Nenad Bursac

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  1. James A. Scull
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  2. Luke C. McSpadden
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  3. Herman D. Himel IV
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  4. Nima Badie
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Correspondence to Nenad Bursac.

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Associate Editor Nathalie Virag oversaw the review of this article.

James A. Scull and Luke C. McSpadden contributed equally to this work.

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Scull, J.A., McSpadden, L.C., Himel, H.D. et al. Single-Detector Simultaneous Optical Mapping of V m and [Ca2+]i in Cardiac Monolayers. Ann Biomed Eng 40, 1006–1017 (2012). https://doi.org/10.1007/s10439-011-0478-z

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