Fluorescent microparticles for sensing cell microenvironment oxygen levels within 3D scaffolds.
Fluorescent microparticles for sensing cell microenvironment oxygen levels within 3D scaffolds.
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DOI：
10.1016/j.biomaterials.2009.02.021
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发表时间：
2009-06
期刊：
影响因子：
14
通讯作者：
Leach, Jennie B.
中科院分区：
文献类型：
--
作者：
Acosta, Miguel A.;Ymele-Leki, Patrick;Kostov, Yordan V.;Leach, Jennie B.
我们提出了荧光氧传感微粒的开发和表征，其设计用于测量标准细胞培养和透明三维（3D）细胞支架内存在的微环境中的氧浓度。微粒合成采用聚（二甲基硅氧烷）来封装与氧敏感发光团以及对氧不敏感的参考或归一化荧光团结合的硅胶颗粒。我们开发了一种基于荧光显微镜的快速，自动化和非侵入性传感器分析方法来测量水凝胶支架中的氧浓度。我们证明，微粒是非细胞毒性的，他们的反应是一个传统的溶解氧计。选择微粒尺寸（5-40 μm）用于氧浓度的微尺度映射，以允许对单个细胞进行局部测量。评估了两种校准方法，并揭示了传感器系统能够表征与细胞和组织生物学相关的缺氧到高氧条件的范围（即，pO2 10-160 mm Hg）。校准分析还显示，微粒具有高比例的淬灭发光体（0.90 ± 0.02），表明所报道的方法为传感器性能提供了显著的优势。因此，这项研究报告了一种多功能的氧传感技术，使未来的相关性的局部氧浓度与个别细胞的反应，在培养的工程组织。
We present the development and characterization of fluorescent oxygen-sensing microparticles designed for measuring oxygen concentration in microenvironments existing within standard cell culture and transparent three-dimensional (3D) cell scaffolds. The microparticle synthesis employs poly(dimethylsiloxane) to encapsulate silica gel particles bound with an oxygen-sensitive luminophore as well as a reference or normalization fluorophore that is insensitive to oxygen. We developed a rapid, automated and non-invasive sensor analysis method based on fluorescence microscopy to measure oxygen concentration in a hydrogel scaffold. We demonstrate that the microparticles are non-cytotoxic and that their response is comparable to that of a traditional dissolved oxygen meter. Microparticle size (5–40 μm) was selected for microscale-mapping of oxygen concentration to allow measurements local to individual cells. Two methods of calibration were evaluated and revealed that the sensor system enables characterization of a range of hypoxic to hyperoxic conditions relevant to cell and tissue biology (i.e., pO2 10–160 mm Hg). The calibration analysis also revealed that the microparticles have a high fraction of quenched luminophore (0.90 ± 0.02), indicating that the reported approach provides significant advantages for sensor performance. This study thus reports a versatile oxygen-sensing technology that enables future correlations of local oxygen concentration with individual cell response in cultured engineered tissues.
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