Evaluating Nanoparticle Sensor Design for Intracellular pH Measurements
Evaluating Nanoparticle Sensor Design for Intracellular pH Measurements
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DOI：
10.1021/nn201643f
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发表时间：
2011-07-01
期刊：
影响因子：
17.1
通讯作者：
Andresen, Thomas L.
中科院分区：
文献类型：
--
作者：
Benjaminsen, Rikke V.;Sun, Honghao;Andresen, Thomas L.
过去十年来，基于颗粒的纳米传感器被设计用于基于光学荧光的比率测量活细胞中的 pH 值。然而，使用颗粒纳米传感器对内涵体和溶酶体中的 pH 值进行定量和时间分辨的细胞内测量具有挑战性，并且需要改进测量方法。在本文中，我们使用纳米颗粒传感器成功地对活细胞内体和溶酶体进行了时间分辨 pH 测量，并展示了传感器选择对于成功定量的重要性。我们研究了两种通过内吞作用内化的基于纳米粒子的传感器系统，并阐明了纳米传感器设计中在未来开发新传感器时应考虑的重要因素。从我们的实验中可以清楚地看出，使用具有广泛测量范围的传感器非常重要，因为当测量 pH 值太接近传感器敏感范围的极限时，错误的 pH 值量化是一个不幸的结果。 pH测量范围为3.2-7.0的三重标记纳米传感器是通过向每个传感器添加两个具有不同pK(a)的pH敏感荧光团而合成的，似乎可以解决在内体-溶酶体途径中测量pH时发现的一些早期问题。
Particle-based nanosensors have over the past decade been designed for optical fluorescent-based ratiometric measurements of pH in living cells. However, quantitative and time-resolved intracellular measurements of pH in endosomes and lysosomes using particle nanosensors are challenging, and there is a need to improve measurement methodology. In the present paper, we have successfully carried out time-resolved pH measurements in endosomes and lyosomes in living cells using nanoparticle sensors and show the importance of sensor choice for successful quantification. We have studied two nanoparticle-based sensor systems that are internalized by endocytosis and elucidated important factors in nanosensor design that should be considered In future development of new sensors. From our experiments it is dear that it is highly Important to use sensors that have a broad measurement range, as erroneous quantification of pH is an unfortunate result when measuring pH too close to the limit of the sensitive range of the sensors. Triple-labeled nanosensors with a pH measurement range of 3.2-7.0, which was synthesized by adding two pH-sensitive fluorophores with different pK(a) to each sensor, seem to be a solution to some of the earlier problems found when measuring pH In the endosome-lysosome pathway.