Spectral evidence for non-calcium interactions of intracellular Indo-1.

Spectral evidence for non-calcium interactions of intracellular Indo-1.
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细胞内 Indo-1 非钙相互作用的光谱证据。

DOI:
10.1016/0006-291x(89)92139-6
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发表时间:
1989
影响因子:
3.1
通讯作者:
R. Shuler
R. Shuler
中科院分区:
生物学4区
文献类型:
--
作者:
C. Owen;R. Shuler

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Indo-1 广泛用于测量细胞内游离钙 [Ca2+]i,通过将 2 个或更多波长处的荧光发射与 Indo-1 在完全结合钙和完全不含钙时的发射(假设已知)进行比较。准确的定量需要在细胞内染料上获得这些“参考”值,并且本研究的全光谱表明,原因是无钙峰的显着光谱偏移,而不是钙结合峰的光谱偏移。数学分析表明,新峰一定是Indo-1分子的新状态,因为它不能简单地归因于细胞中残留的钙。当细胞内“参考”光谱用于数据分析时,[Ca2+]i 可以从整个光谱或两个波长下观察到的比率计算得出,并且具有良好的一致性。当使用细胞外“参考”光谱时,通过比率方法计算的值取决于波长的选择。
Indo-1 is widely used to measure intracellular free calcium, [Ca2+]i, by comparing the fluorescence emission at 2 or more wavelengths with the emissions, which are assumed to be known, of Indo-1 when it is fully calcium-bound and when it is fully calcium-free. Accurate quantitation requires that these “reference” values be obtained on intracellular dye, and the full spectra of this study show that the reason is a significant spectral shift of the calcium-free peak, but not the calcium-bound. A mathematical analysis shows that the new peak must be a new state of the Indo-1 molecule, since it cannot be simply due to residual calcium in the cell. When intracellular “reference” spectra were used in the data analysis, [Ca2+]icould be calculated from whole spectra or from the ratio of observations at two wavelengths with good agreement. When extracellular “reference” spectra were used, the value calculated by the ratio method depended on the choice of wavelengths.
DOI: --
发表时间: 1986
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影响因子: --
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发表时间: 1986-08
影响因子: 4.4
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