Skip to main content
SpringerLink
Log in
Book cover

Cell Membrane Transport pp 257–272Cite as

A Practical Introduction to the Use of Intracellular Fluorescent Indicators

  • Chapter

  • 108 Accesses

Abstract

The concentrations of intracellular ions such as Cat-’, Na+ and H+ are important in the regulation of cell function. Much effort has therefore been expended in measuring them. However, until the last few years, most direct methods for measurement of these concentrations were only suitable for large cells. The situation has changed completely with the introduction of fluorescent indicators. Here we describe the properties of some of these indicators.

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

Chapter
USD   29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   169.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   219.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Almers, W. and Neher, E. (1985). The calcium signal from Fura-2 loaded mast cells depends strongly on the method of dye loading. FEBS Letters 192, 13–18

    Article  PubMed  CAS  Google Scholar 

  • Arslan, P.,De Virgilio, R. Y., Tsien, R. Y., and Pozzan, T.(1984). Cytosolic Ca2+ homeostasis in Ehrlich and Yoshida carcinomas. A new membrane-permeant chelator of heavy metals reveals that these ascite tumor cells have normal cytosolic free Ca2+. J. Biol. Chem. 260,2719–2727.

    Google Scholar 

  • Albert, P. R., and Tashjian, A. H. (1984).Thyrotropin-releasing hormone-induced spike and plateau in cytosolic free Ca2+ concentration in pituitary cells. Relation to prolactin release. J. Biol. Chem. 259, 5827–5832.

    Google Scholar 

  • Borle, A. B., and Snowdowne, K. W. (1986). Methods for the measurement of intracellular ionized calcium in mammalian cells: comparison of four classes of calcium indicators. In Calcium and cell function ed Cheung, W.Y. vol 7, pp 159–200. Orlando: Academic Press.

    Google Scholar 

  • Becker, P. L., and Fay.,F. (1987). Photobleaching of fura-2 and its effects on determination of calcium concentrations. Am. J. Physiol. 253, C613–C61ß.

    Google Scholar 

  • du Bell, W. H., Philips, Ch. P., and Houser, S. R. (1988). A technique for measuring cytosolic free Ca2+ with Indo-1 in feline myocytes. in “Biology of isolated adult cardiac myocytes” eds Clark, W.A., Decker, R.S. & Borg, T.K. Elsevier Science Publishing Co.

    Google Scholar 

  • Chaillet, J.R. & Boron, W.F. (1985). Intracellular calibration of a pH-sensitive dye in isolated, perfused salamander proximal tubules. J. gen. Physiol. 86, 765–794.

    Google Scholar 

  • Chance, B., Cohen, P., Jobsis, F. & Schoener, B. (1962). Intracellular oxidation-reduction states in vivo. Science (Wash D.C.) 137, 499–508.

    Article  CAS  Google Scholar 

  • Cobbold, P. H. and Rink, T. J. (1987). Fluorescence and bioluminescence measurements of cytoplasmic free calcium. Biochem. J. 248, 313–328.

    Google Scholar 

  • Davis, M. H., Altschuld, R. A., Jung, D. W., and Brierley, G. P., (1987). Estimation of intramitochondrial pCa nd pH by fura-2 and 2,7 biscarboxyethyl-5(6)-carboxyfluorescein (BCECF) fluorescence. Biochem. Biophys. Res. Comm. 149, 40–45.

    Google Scholar 

  • Deber, C. M., Tom-Kun, J., Mack, E., and Grinstein, S. (1985). Bromo A23187: a fluorescent calciun ionophore for the use with fluorescent probes. Anal. Biochem. 146, 349–352.

    Google Scholar 

  • De feo, T. T., Briggs, G. M. and Morgan, K. G. (1987). CaZ+ signals obtained with multiple indicators in mammalian vascular muscle cells. Am. J.Physiol. 253, H1456–H1461.

    Google Scholar 

  • Donoso, P. Eisner, D.A. & O’Neill, S.C. (1989a). Measurement of intracellular [Na+] in isolated rat cardiac myocytes using the fluorescent indicator SBFI. J. Physiol. 418, 48 P.

    Google Scholar 

  • Donoso, P., Eisner, D.A., O’Neill, S.C. & Valdeolmillos, M. (1989b). the fluorescent probe Indo-1 can indicate [CaZ+]i and caffeine concentration simultaneously. J. Physiol. 416, 42P.

    Google Scholar 

  • Eisner, D. A., Kenning, N. A., O’Neill, S. C., Pocok, G., Richards, C. D., and Valdeolmillos, M., (1989a). A novel method for absolute calibration of intracellular pH indicators. Pflugers Arch. 413, 145–157.

    Google Scholar 

  • Eisner, D. A., Nichols, C. G., O’Neill, S. C., Smith, G. L., and Valdeolmillos, M., (1989b). The effects of metabolic inhibition on intracellular calcium and pH in isolated rat ventricular myocytes. J. Physiol. 411, 393–418.

    PubMed  CAS  Google Scholar 

  • Eng, J., Lynch, R. M., and Balaban, R. S. (1989). Nicotinamide adenine dinucleotide fluorescence spectroscopy and imaging of isolated cardiac myocytes. Biophys. J. 55, 621–630.

    Google Scholar 

  • Garcia-Sancho, J. (1984). Inhibition of glycolysis in the human erytrocyte by formaldehyde and Ca-chelator esters, J. Physiol. 357, 99.

    Google Scholar 

  • Grynkiewicz, G., Ponie, M., and Tsien, R. Y. (1985). A new generation of Ca2+ indicators with greatly improved fluorescence properties. J. Biol. Chem. 260, 3440–3450.

    Google Scholar 

  • Gunter, T. H., Restrepo, D., and Gunter, K. K. (1988). Conversion of esterified fura-2 and indo-1 to CaZ+ sensitive forms by mitochondria. Am. J. Physiol. 255, C304–C310.

    Google Scholar 

  • Hesketh, T.R., Smith, G.A., Moore, J.P., Taylor, M.V. & Metcalfe, J.C. (1983). Free cytoplasmic calcium concentration and the m i t o g e n i c stimulation of lymphocytes. J. biol. Chem. 258, 4876–4882.

    Google Scholar 

  • Illsley, N.P. & Verkman, A.S. (1987). Membrane chloride transport censured using a chloride-sensitive fluorescent probe. Biochemistry 26, 1215–1219

    CAS  Google Scholar 

  • Johnson, P. C., Ware, J. A., Cliveden, P. B., Smith, H., Dvorak, A. H., and Salzman, E. W. (1985). Measurement of ionized calcium in blood platelets with the photoprotein aequorin. Comparison with quin-2. J. Biol. Chem. 260, 2069–2076.

    Google Scholar 

  • Knight, D. E., and Kesteven, N. T. (1983). Evoked transient, intracellular free CaZ+ changes and secretion in isolated bovine adrenal medullary cells. Proc. R. Soc. London Ser. B 218, 177–199.

    Google Scholar 

  • Konishi, M., Olson, A., Hollingworth, S., and Baylor, S. M. (1988). Myoplasmic binding of fura-2 investigated by steady-state fluorescence and absorbance measurements. Biophys. J. 54, 1089–1104.

    Google Scholar 

  • Kurtz, I. & Balaban, R.S. (1985). Fluorescence emission spectroscopy of 1,4-dihydroxyphthalonitrile. A method for determining intracellular pH ni cultured cells. Biophys. J. 48, 499–508.

    Google Scholar 

  • Lakowicz, J. R. (1983). “Principles of fluorescence spectroscopy” Plenum Press N.Y.

    Google Scholar 

  • Li, Q., Altschuld, R.A. & Stokes, B.T. (1987). Quantitation of intracellular free calcium in single adult cardiomyocytes by Fura-2 fluorescence microscopy: calibration of fura-2 ratios. Biochem. Biophys. res. Comm. 147, 120–126.

    Google Scholar 

  • Macintyre, E.A., Tatham, P.E.R., Abdul-Gaffar, R. & Linch, D.C. (1988). The effects of pertussis toxin on human T lymphocytes. Immunology 64, 427432.

    Google Scholar 

  • Minta, A., Kao, J.P. & Tsien, R.Y. (1989). Fluorescent indicators for cytosolic calcium based on rhodamine and fluorescein chromophores..J biol. Chem. 264, 8171–8178.

    Google Scholar 

  • Minta, A. & Tsien, R.Y. (1989). Fluorescent indicators for cytosolic sodium J. biol. Chem. 264, 19449–19457

    Google Scholar 

  • Moore, E.D.W., Tsien, R.Y., Minta, A. & Fay, F.S. (1988). Measurement of intracellular sodium with SBFP. A newly developed sodium sensitive fluorescent dye. FASEB J. 2, A754.

    Google Scholar 

  • Oakes, S. G., Martin, W. J., Lisek,. A., and Powis, G. (1988). Incomplete hydrolysis of the calcium indicator precursor Fura-2 pentaacetoxymethyl ester (Fura-2 AM) by cells. Anal. Biochem. 169, 159–166.

    CAS  Google Scholar 

  • Owen, Ch. S. (1988). Quantitation of lymphocyte intracellular free calcium signals using indo-1. Cell Calcium 9, 141–149.

    CAS  Google Scholar 

  • Popov, E. G., Gavrilov, I. Yu., Pozin, E. Ya., and Gabbasov, Z. A. (1988). Multiwavelength method for measuring concentration of free cytosolic calcium using the fluorescent probe indo-1. Arch. Biochem. Biophys. 261, 91–96.

    Google Scholar 

  • Rink, T. J., and Pozzan, T. (1985). Using Quin-2 in cell suspensions. Cell Calcium 6, 133–144.

    Article  CAS  Google Scholar 

  • Rink, T. J., Tsien, R. Y., and Pozzan, T. (1982). Cytoplasmic pH and Mgt+ ni lymphocytes. J. Cell. Biol. 92, 189–196.

    Google Scholar 

  • Szatkowski, M. S., and Thomas, R. C. (1986). New method for calculating pH, from accurately measured changes in pH; induced by a weak acid and base. Pflugers Arch. 407, 59–63.

    CAS  Google Scholar 

  • Thomas, J.A., Buchsbaum, R.N., Zimniak, A. & Racker, E. (1979). Intracellular pH measurements in Ehrlich ascites tumour cells utilizing spectroscopic probes generated in situ. Biochemistry 18, 2210–2218.

    CAS  Google Scholar 

  • Timmerman, M. P., and Ashley, C. C. (1986). Fura-2 difussion and its use sa an indicator of transient free calcium changes in single striated muscle cells. FEBS Lett. 209, 1–8.

    CAS  Google Scholar 

  • Tsien, R. Y. (1988). Fluorescence measurement and photochemical manipulation of cytosolic free calcium. Trends Neuros. 11, 419–424.

    CAS  Google Scholar 

  • Tsien, R.Y. & Minta, A. (1987). J. cell. Biol. 105 89a

    Google Scholar 

  • Tsien, R. Y., Pozzan, T., and Rink, T. J. (1982). Calcium homeostasis in intact lymphocytes: cytoplsmic free calcium monitored with a new intracellularly trapped fluorescent indicator. J. Cell Biol. 94,325–334.

    Google Scholar 

  • Tsien, R. Y., Rink, T. J., and Poenie, M. (1985). Measurement of cytosolyc free calcium Cat+ in individual small cells using fluorescence microscopy with dual excitation wavelengths. Cell Calcium 6, 145–157.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Departamento de Fisiologia, Facultad Medicina, Universidad de Alicante, Spain

    M. Valdeolmillos

  2. Department of Physiology, University College London, Gower St., London, WC1E 6BT, Great Britain

    D. A. Eisner

Authors
  1. M. Valdeolmillos
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D. A. Eisner
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. University of London, London, UK

    David L. Yudilevich

  2. University of Chile, Santiago, Chile

    Rosa Devés

  3. University of Málaga, Málaga, Spain

    Salvador Perán

  4. The Hebrew University of Jerusalem, Jerusalem, Israel

    Z. Ioav Cabantchik

Rights and permissions

Reprints and permissions

Copyright information

© 1991 Springer Science+Business Media New York

About this chapter

Cite this chapter

Valdeolmillos, M., Eisner, D.A. (1991). A Practical Introduction to the Use of Intracellular Fluorescent Indicators. In: Yudilevich, D.L., Devés, R., Perán, S., Cabantchik, Z.I. (eds) Cell Membrane Transport. Springer, Boston, MA. https://doi.org/10.1007/978-1-4757-9601-8_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-4757-9601-8_14

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4757-9603-2

  • Online ISBN: 978-1-4757-9601-8

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation