Comparison of 4-Mercaptobenzoic Acid Surface-Enhanced Raman Spectroscopy-Based Methods for pH Determination in Cells.
Comparison of 4-Mercaptobenzoic Acid Surface-Enhanced Raman Spectroscopy-Based Methods for pH Determination in Cells.
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基于细胞中pH测定的基于4-羟基苯甲酸表面增强拉曼光谱法的比较。
DOI：
10.1177/0003702820950768
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发表时间：
2020-11
影响因子：
3.5
通讯作者：
Schultz ZD
中科院分区：
文献类型：
--
作者：
Scarpitti BT;Morrison AM;Buyanova M;Schultz ZD
细胞pH值的测量用于推断信息，如细胞周期的阶段，癌症和其他疾病的存在，以及治疗药物的递送或效果。基于纳米颗粒的pH探针的表面增强拉曼光谱（Sers）已被用于询问细胞内pH，与荧光指示剂相比具有避免光漂白的显著优点。4-巯基苯甲酸（MBA）是一种常用的pH敏感报告分子。通过表面增强拉曼光谱进行细胞内pH传感需要分析所观察到的MBA光谱，并且光谱干扰会影响pH测定。来自普通细胞容器的背景、成像太少的颗粒、信噪比和报告分子的降解是可能改变细胞中适当的基于SERS的pH测定的因素之一。在这里，我们比较了常见的光谱分析方法，看看不同的因素如何改变计算的pH值在拉曼地图MBA功能化Au nanostars在SW 620癌细胞。我们比较的方法包括使用ν（COO−）伸缩的相对强度，ν8a模式的化学计量学分析，以及分析ν8a模式的频移。这些方法对活细胞实验中的这些误差来源中的一些显示出不同的灵敏度。基于拉曼频移的pH测定似乎给出了更可靠的pH测定，尽管在高信噪比环境中，强度比可以为细胞成像提供对pH的小变化的更好的灵敏度。
Measurements of cellular pH are used to infer information such as stage of cell cycle, presence of cancer and other diseases, as well as delivery or effect of a therapeutic drug. Surface enhanced Raman spectroscopy (SERS) of nanoparticle-based pH probes have been used to interrogate intracellular pH, with the significant advantage of avoiding photobleaching compared to fluorescent indicators. 4-mercaptobenzoic acid (MBA) is a commonly used pH sensitive reporter molecule. Intracellular pH sensing by surface enhanced Raman spectroscopy requires analysis of the observed MBA spectrum and spectral interference can affect the pH determination. Background from common cell containers, imaging too few particles, signal to noise ratios, and degradation of reporter molecules are among the factors that may alter appropriate SERS-based pH determination in cells. Here, we have compared common methods of spectral analysis to see how different factors alter the calculated pH in Raman maps of MBA functionalized Au nanostars in SW620 cancer cells. The methods included in our comparison use the relative intensity of the ν(COO−) stretch, chemometric analysis of the ν8a mode, and analyzing the frequency shift of the ν8a mode. These methods show different sensitivity to some of these sources of error in live cell experiments. pH determination based on Raman frequency shift appears to give a more reliable pH determination, though in high signal to noise environments, intensity ratios may provide better sensitivity to small changes in pH for cellular imaging.
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影响因子：
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作者：
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DOI：
10.3390/s131216736
发表时间：
2013-12-05
期刊：
Sensors (Basel, Switzerland)
影响因子：
--
作者：
Benčina M
通讯作者：
Benčina M
影响因子：
3.7
作者：
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通讯作者：
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