PMID: 9160163Apr 1, 1997Paper

Nucleoplasmic and cytoplasmic differences in the fluorescence properties of the calcium indicator Fluo-3

Cell Calcium
C Perez-TerzicD E Clapham

Abstract

The fluorescent indicator Fluo-3 is widely used to monitor the calcium concentration ([Ca2+]) in the cytoplasm and nucleus of various cells. Estimates of nuclear [Ca2+] are based on the assumption of identical behavior of Fluo-3 in different cellular compartments. The assumption is not valid if the fluorescence properties of the dye are altered by the nuclear environment, independent of the [Ca2+]. To determine the effects of the nucleoplasm on the behavior of Fluo-3, we applied laser scanning confocal microscopy and spectrophotometry to measure fluorescence intensity as well as emission and absorbance spectra of the Ca2+ indicator, Fluo-3. Spectra were measured in intact Xenopus oocytes, neuroblastoma cells, and cytoplasmic and nucleoplasmic homogenates. The fluorescence signal in intact cells loaded with Fluo-3 was approximately 2-times higher in the nucleus when compared to the cytoplasm. The fluorescence intensity of Fluo-3 in nucleoplasmic homogenates was higher than in cytoplasmic homogenates or internal buffers even when [Ca2+] was clamped. Despite identical [Ca2+], pH, and temperature, the emission and absorbance spectra of Fluo-3 from nuclear homogenates displayed a higher fluorescence at each wavelength measured when ...Continue Reading

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Citations

Sep 17, 2005·Journal of Bioenergetics and Biomembranes·Antonio LaxFrancisco Fernández-Belda
Jan 16, 2003·Cell Calcium·Masumi InoueAkira Warashina
May 28, 1998·Cell Calcium·M N BadmintonA K Campbell
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