NanoPolyPhotonik, Fraunhofer Institute for Applied Polymer Research, Potsdam-Golm, Germany.
Sensors (Basel). 2012;12(6):7015-32. doi: 10.3390/s120607015. Epub 2012 May 25.
Pioneered by Clark's microelectrode more than half a century ago, there has been substantial interest in developing new, miniaturized optical methods to detect molecular oxygen inside cells. While extensively used for animal tissue measurements, applications of intracellular optical oxygen biosensors are still scarce in plant science. A critical aspect is the strong autofluorescence of the green plant tissue that interferes with optical signals of commonly used oxygen probes. A recently developed dual-frequency phase modulation technique can overcome this limitation, offering new perspectives for plant research. This review gives an overview on the latest optical sensing techniques and methods based on phosphorescence quenching in diverse tissues and discusses the potential pitfalls for applications in plants. The most promising oxygen sensitive probes are reviewed plus different oxygen sensing structures ranging from micro-optodes to soluble nanoparticles. Moreover, the applicability of using heterologously expressed oxygen binding proteins and fluorescent proteins to determine changes in the cellular oxygen concentration are discussed as potential non-invasive cellular oxygen reporters.
半个多世纪前，Clark 的微电极开创了先河，此后人们一直热衷于开发新的、微型化的光学方法来检测细胞内的分子氧。虽然这种方法在动物组织测量中得到了广泛应用，但在植物科学中，细胞内光学氧生物传感器的应用仍然很少。一个关键的方面是绿色植物组织强烈的自发荧光会干扰常用氧探针的光学信号。最近开发的双频相位调制技术可以克服这一限制，为植物研究提供了新的视角。本文综述了基于不同组织中磷光猝灭的最新光学传感技术和方法，并讨论了在植物中应用的潜在问题。本文还综述了最有前途的氧敏感探针，以及从微光纤到可溶性纳米颗粒等不同的氧传感结构。此外，还讨论了异源表达的氧结合蛋白和荧光蛋白在确定细胞氧浓度变化方面的应用，作为潜在的非侵入性细胞氧报告器。
