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首页> 外文期刊>Journal of Neuroscience Methods >Intracellular ionic concentration by calibration from fluorescence indicator emission spectra, its relationship to the K(d), F(min), F(max) formula, and use with Na-Green for presynaptic sodium.
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Intracellular ionic concentration by calibration from fluorescence indicator emission spectra, its relationship to the K(d), F(min), F(max) formula, and use with Na-Green for presynaptic sodium.

机译：通过荧光指示剂发射光谱的校准对细胞内离子浓度进行校准，确定其与K（d），F（min），F（max）公式的关系，并与Na-Green一起用于突触前钠。

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The emission spectra calibration curves for a fluorescence indicator and the F(min), F(max), and K(d) formula were shown to be related. Using the known calibrated fluorescence emitted by Sodium Green (Na-Green) and photo-multiplier-tube quantum efficiency, we calculated the detection signal over a range of sodium concentrations. The calculated calibration curves were compared for optical filters passing a narrow band, medium band or full spectrum. We found that a method based on the full emission spectrum was the most appropriate. Given a known resting concentration of intracellular sodium, calibrated readings can be converted to concentration values. This method is applicable to any fluorescence indicator when curves for emission spectra over a range of concentrations are available. We measured sodium concentration changes during trains of action potentials (APs) at a crayfish motor axon's presynaptic terminals injected with Na-Green. During low frequency AP trains, net sodium increases asymptotically with frequency. Average net Na-flux per AP decreases for increasing terminal size. The terminals of crayfish motor axon have surface area to volume ratio which is 7700 times larger than for squid. Thus, in comparison to squid, crayfish terminals exhibit a larger change in [Na(+)](i) during equivalent AP activity.
机译：荧光指示剂的发射光谱校准曲线与F（min），F（max）和K（d）公式相关。使用由钠绿色（Na-Green）发射的已知校准荧光和光电倍增管量子效率，我们计算了钠浓度范围内的检测信号。比较了通过窄带，中带或全光谱的光学滤波器的计算出的校准曲线。我们发现，基于全发射光谱的方法是最合适的。给定已知的细胞内钠静息浓度，可以将校准后的读数转换为浓度值。当可以获得一系列浓度范围内的发射光谱曲线时，该方法适用于任何荧光指示剂。我们测量了在小龙虾运动轴突突触前末端注射Na-Green的一系列动作电位（APs）中钠浓度的变化。在低频AP训练期间，净钠随频率渐近增加。随着终端尺寸的增加，每个AP的平均净Na-通量减少。小龙虾运动轴突的末端的表面积与体积之比为鱿鱼的7700倍。因此，与鱿鱼相比，小龙虾末端在同等的AP活性下表现出[Na（+）]（i）较大的变化。

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