Skip to main content
Log in

A method for recording intracellular [Ca2+] transients in cardiac myocytes using calcium green-2

  • Original Article
  • Heart, Circulation, Respiration and Blood; Enviromental and Exercise Physiology
  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

Calcium green-2 (Ca green) is a non-ratio-metric fluorescent Ca2+ indicator with an affinity for Ca2+ (dissociation constant Kd=3 μM) that is lower than more commonly used indicators such as fura-2 and fluo-3. This low Ca2+ affinity, coupled with a high quantum yield, allows cells to be loaded with low concentrations of Ca green, avoiding problems of cytosolic Ca2+ buffering and a low signal-to-noise ratio. This communication presents a method for monitoring intracellular [Ca2+] changes in isolated rat ventricular myocytes loaded with Ca green and the fluorescent pH indicator carboxy SNARF-1 (SNARF). SNARF provides a Ca2+-insensitive signal with which Ca green fluorescence can be corrected for cell motion and dye-loading artifacts.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Backx PH, ter Keurs HEDJ (1993) Fluorescent properties of rat cardiac trabeculae micro-injected with fura-2 salt. Am J Physiol 264:H1098–1110

    PubMed  Google Scholar 

  2. Balke CW, Egan TM, Wier WG (1994) Processes that remove calcium from the cytoplasm during excitation-contraction coupling in intact rat heart cells. J Physiol (Lond) 474:447–462

    Google Scholar 

  3. Baylor SM, Hollingworth S (1988) Fura-2 calcium transients in frog skeletal muscle fibres. J Physiol (Lond) 403:151–192

    Google Scholar 

  4. Baylor SM, Harkins AB, Kurebayashi N (1994) Letter in reply to Westerblad and Alien. Biophys J 66:927–928

    Google Scholar 

  5. Berlin JR, Konishi M (1993) Ca2+ transients in cardiac myocytes measured with high and low affinity Ca2+ indicators. Biophys 165:1632–1647

    Google Scholar 

  6. Bers DM, Lederer WJ, Berlin JR (1990) Intracellular Ca transients in rat cardiac myocytes: role of Na-Ca exchange in excitation-contraction coupling. Am J Physiol 258:C944-C954

    PubMed  Google Scholar 

  7. Blatter LA, McGuigan JA (1991) Intracellular pH regulation in ferret ventricular muscle: the role of Na-H exchange and the influence of metabolic substrates. Circ Res 68:150–161

    PubMed  Google Scholar 

  8. Cannell MB, Berlin JR, Lederer WJ (1987) Effect of membrane potential changes on the calcium transient in single rat cardiac muscle cells. Science 238:1419–1423

    PubMed  Google Scholar 

  9. Cleemann L, Morad M (1991) Role of Ca2+ channel in cardiac excitation-contraction coupling in the rat: evidence from Ca2+ transients and contraction. J Physiol (Lond) 432:283–312

    Google Scholar 

  10. Hove-Madsen L, Bers DM (1992) Indo-1 binding to protein in permeabilized ventricular myocytes alters its spectral and Ca binding properties. Biophys J 63:89–97

    PubMed  Google Scholar 

  11. Konishi M, Berlin JR (1993) Ca transients in cardiac myocytes measured with a low affinity fluorescent indicator, furaptra. Biophys J 64:1331–1343

    PubMed  Google Scholar 

  12. Lipp P, Niggli E (1993) Microscopic spiral-waves reveal positive feedback in subcellular calcium signaling. Biophys J 65:2272–2276

    PubMed  Google Scholar 

  13. Lipp P, Niggli E (1993) Ratiometric confocal Ca2+-measurements with visible wavelength indicators in isolated cardiac myocytes Cell Calcium 14:359–372

    Article  PubMed  Google Scholar 

  14. Rijkers GT, Justement LB, Griffioen AW, Cambier JC (1990) Improved method for measuring intracellular Ca2+ with fluo-3. Cytometry 11:923–927

    PubMed  Google Scholar 

  15. Westerblad H, Allen DG (1994) Methods for calibration of fluorescent calcium indicators in skeletal muscle fibers. Biophys J 66:926–927

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Spencer, C.I., Berlin, J.R. A method for recording intracellular [Ca2+] transients in cardiac myocytes using calcium green-2. Pflugers Arch. 430, 579–583 (1995). https://doi.org/10.1007/BF00373895

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00373895

Key words

Navigation