Fluorescence lifetime microscopy of the sodium indicator sodium-binding benzofuran isophthalate in HeLa cells

Sanda Despa, Paul Steels, Marcel Ameloot

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The behavior of the sodium indicator sodium-binding benzofuran isophthalate (SBFI) is investigated in HeLa cells by time-resolved fluorescence microscopy. The fluorescence relaxation of SBFI in HeLa cells can be described by a triexponential for intracellular sodium concentration ([Na+](i)) between 0 and 90 mM. Changes in [Na+](i) affect neither the fluorescence relaxation times (0.21, 0.60, and 2.7 ns) nor the average decay time (2.2 ns). The preexponential factor of the shortest decay time is negative. However, the ratio of the fluorescence excitation signal at 340 nm to that at 380 nm increases with [Na+](i). To elucidate the behavior of SBFI in cells, experiments are performed on SBFI in buffer at various concentrations of sodium, potassium, and bovine serum albumin (BSA) and at various viscosities. The fluorescence decay is triexponential only in the presence of BSA. The relaxation times are independent of [Na+] and [BSA]. The preexponential factor of the shortest decay time is negative from a certain [BSA] on, which depends on [Na+]. The data indicate that interactions with intracellular components rather than microviscosity influence the SBFI behavior in cells. A model is suggested in which the fluorescence intensities are mainly determined by the signals from the Na+ subset of SBFI and SBFI subset of protein complexes. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)227-241
Number of pages15
JournalAnalytical Biochemistry
Volume280
Issue number2
DOIs
StatePublished - May 1 2000
Externally publishedYes

Fingerprint

HeLa Cells
Fluorescence Microscopy
Microscopic examination
Sodium
Fluorescence
Bovine Serum Albumin
Relaxation time
Fluorescence microscopy
sodium-binding benzofuran isophthalate
Viscosity
Potassium
Buffers
Proteins
Experiments

Keywords

  • Bovine serum albumin
  • HeLa cells
  • SBFI
  • Time-resolved fluorescence microscopy

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Fluorescence lifetime microscopy of the sodium indicator sodium-binding benzofuran isophthalate in HeLa cells. / Despa, Sanda; Steels, Paul; Ameloot, Marcel.

In: Analytical Biochemistry, Vol. 280, No. 2, 01.05.2000, p. 227-241.

Research output: Contribution to journalArticle

Despa, Sanda ; Steels, Paul ; Ameloot, Marcel. / Fluorescence lifetime microscopy of the sodium indicator sodium-binding benzofuran isophthalate in HeLa cells. In: Analytical Biochemistry. 2000 ; Vol. 280, No. 2. pp. 227-241.
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