Effects of temperature on calcium-sensitive fluorescent probes

Ann E. Oliver, Gary A. Baker, Robert D. Fugate, Fern Tablin, John H. Crowe

Research output: Contribution to journalArticle

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Abstract

The effect of temperature on the binding equilibria of calcium-sensing dyes has been extensively studied, but there are also important temperature- related changes in the photophysics of the dyes that have been largely ignored. We conducted a systematic study of thermal effects on five calcium- sensing dyes under calcium-saturated and calcium-free conditions. Quin-2, chlortetracycline, calcium green dextran, Indo-1, and Fura-2 all show temperature-dependent effects on fluorescence in all or part of the range tested (5-40°C). Specifically, the intensity of the single-wavelength dyes increased at low temperature. The ratiometric dyes, because of variable effects at the two wavelengths, showed, in general, a reduction in the fluorescence ratio as temperature decreased. Changes in viscosity, pH, oxygen quenching, or fluorescence maxima could not fully explain the effects of temperature on fluorescence. The excited-state lifetimes of the dyes were determined, in both the presence and absence of calcium, using multifrequency phase-modulation fluorimetry. In most cases, low temperature led to prolonged fluorescence lifetimes. The increase in lifetimes at reduced temperature is probably largely responsible for the effects of temperature on the physical properties of the calcium-sensing dyes. Clearly, these temperature effects can influence reported calcium concentrations and must therefore be taken into consideration during any investigation involving variable temperatures.

Original languageEnglish (US)
Pages (from-to)2116-2126
Number of pages11
JournalBiophysical Journal
Volume78
Issue number4
StatePublished - Apr 2000

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Fluorescent Dyes
Calcium
Temperature
Coloring Agents
Fluorescence
Chlortetracycline
Fluorometry
Fura-2
Viscosity
Hot Temperature
Oxygen

ASJC Scopus subject areas

  • Biophysics

Cite this

Oliver, A. E., Baker, G. A., Fugate, R. D., Tablin, F., & Crowe, J. H. (2000). Effects of temperature on calcium-sensitive fluorescent probes. Biophysical Journal, 78(4), 2116-2126.

Effects of temperature on calcium-sensitive fluorescent probes. / Oliver, Ann E.; Baker, Gary A.; Fugate, Robert D.; Tablin, Fern; Crowe, John H.

In: Biophysical Journal, Vol. 78, No. 4, 04.2000, p. 2116-2126.

Research output: Contribution to journalArticle

Oliver, AE, Baker, GA, Fugate, RD, Tablin, F & Crowe, JH 2000, 'Effects of temperature on calcium-sensitive fluorescent probes', Biophysical Journal, vol. 78, no. 4, pp. 2116-2126.
Oliver AE, Baker GA, Fugate RD, Tablin F, Crowe JH. Effects of temperature on calcium-sensitive fluorescent probes. Biophysical Journal. 2000 Apr;78(4):2116-2126.
Oliver, Ann E. ; Baker, Gary A. ; Fugate, Robert D. ; Tablin, Fern ; Crowe, John H. / Effects of temperature on calcium-sensitive fluorescent probes. In: Biophysical Journal. 2000 ; Vol. 78, No. 4. pp. 2116-2126.
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