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Measurement of intracellular Ca2+ concentration using Indo-1 during simultaneous flash photolysis to release Ca2+ from DM-nitrophen

  • Instruments and Techniques
  • Heart, Circulation, Respiration and Blood; Environmental and Exercise Physiology
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Abstract

We have constructed a modular instrument to measure intracellular [Ca2+] ([Ca2+]i) in single isolated cells while simultaneously imposing step changes in [Ca2+]i using “caged Ca2+”. By combining the outputs of a xenon arc lamp with a frequency-tripled (Nd: YAG) laser, the instrument can operate with low maintained illumination to measure [Ca2+]i using a ratiometric Ca2+-sensitive fluorophore and also activate the release of Ca2+ from a caged-Ca2+ compound with a high energy pulse of ultraviolet light. This instrument is simple to assemble, introduces little electrical noise, provides a wide range of illumination power, produces only moderate photobleaching of the Ca2+ indicator and can be readily adapted to diverse cellular preparations. We demonstrate the use of this system to measure step changes in [Ca2+]i in adult rat ventricular myocytes and a human embryonic kidney cell line (293 cells) in culture.

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Authors and Affiliations

  1. Department of Physiology, University of Maryland at Baltimore, 655 West Baltimore St., 21201, Baltimore, MD, USA

    Mark S. Kirby, Robert W. Hadley & W. J. Lederer

  2. Medical Biotechnology Center, University of Maryland at Baltimore, 618 West Lombard St., 21201, Baltimore, MD, USA

    W. J. Lederer

  3. Department of Pharmacology, University of Kentucky, 40536, Lexington, Kentucky, USA

    Robert W. Hadley

Authors
  1. Mark S. Kirby
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  2. Robert W. Hadley
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  3. W. J. Lederer
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Kirby, M.S., Hadley, R.W. & Lederer, W.J. Measurement of intracellular Ca2+ concentration using Indo-1 during simultaneous flash photolysis to release Ca2+ from DM-nitrophen. Pflügers Arch. 427, 169–177 (1994). https://doi.org/10.1007/BF00585957

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  • DOI: https://doi.org/10.1007/BF00585957

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