Accurate Quantitative Sensing of Intracellular pH based on Self-ratiometric Upconversion Luminescent Nanoprobe.
Accurate Quantitative Sensing of Intracellular pH based on Self-ratiometric Upconversion Luminescent Nanoprobe.
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基于自比例上转换发光纳米探针的细胞内 pH 值精确定量检测
DOI：
10.1038/srep38617
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发表时间：
2016-12-09
影响因子：
4.6
通讯作者：
Kong X
中科院分区：
文献类型：
--
作者：
Li C;Zuo J;Zhang L;Chang Y;Zhang Y;Tu L;Liu X;Xue B;Li Q;Zhao H;Zhang H;Kong X
细胞内pH（pHi）的准确定量对揭示细胞活性和疾病预警具有重要意义。已经开发了一系列由不同纳米颗粒或/和染料对组成的基于荧光的纳米生物探针用于pHisensing。目前，生物自发荧光背景和染料的严重光漂白是影响pHi准确定量检测的两个主要因素。在此，我们开发了一种基于Förster共振能量转移（FRET）的自比率发光纳米探针，用于探测pHi，其中pH敏感的异硫氰酸荧光素（FITC）和上转换纳米颗粒（UCNPs）分别作为能量受体和供体。在980 nm激发下，NaYF 4：Yb 3+，Tm 3 +UCNPs在475 nm和645 nm处的上转换发射带分别用作pH响应和自比率参考信号。这种直接定量感测方法规避了传统的基于软件的图像后续处理，这可能导致结果的相对较大的不确定性。由于高效的FRET和无荧光背景，实现了高灵敏度和准确的传感，其特征在于在pH值3.0-7.0的每单位变化3.56，偏差小于0.43。这一方法将有助于pH相关领域的研究和细胞内给药系统的开发。
Accurate quantitation of intracellular pH (pHi) is of great importance in revealing the cellular activities and early warning of diseases. A series of fluorescence-based nano-bioprobes composed of different nanoparticles or/and dye pairs have already been developed for pHisensing. Till now, biological auto-fluorescence background upon UV-Vis excitation and severe photo-bleaching of dyes are the two main factors impeding the accurate quantitative detection of pHi. Herein, we have developed a self-ratiometric luminescence nanoprobe based on förster resonant energy transfer (FRET) for probing pHi, in which pH-sensitive fluorescein isothiocyanate (FITC) and upconversion nanoparticles (UCNPs) were served as energy acceptor and donor, respectively. Under 980 nm excitation, upconversion emission bands at 475 nm and 645 nm of NaYF4:Yb3+, Tm3+UCNPs were used as pHiresponse and self-ratiometric reference signal, respectively. This direct quantitative sensing approach has circumvented the traditional software-based subsequent processing of images which may lead to relatively large uncertainty of the results. Due to efficient FRET and fluorescence background free, a highly-sensitive and accurate sensing has been achieved, featured by 3.56 per unit change in pHivalue 3.0–7.0 with deviation less than 0.43. This approach shall facilitate the researches in pHirelated areas and development of the intracellular drug delivery systems.
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