A novel planar optical sensor for simultaneous monitoring of oxygen, carbon dioxide, pH and temperature

The first quadruple luminescent sensor is presented which enables simultaneous detection of three chemical parameters and temperature. A multi-layer material is realized and combines two spectrally independent dually sensing systems. The first layer employs ethylcellulose containing the carbon dioxide sensing chemistry (fluorescent pH indicator 8-hydroxy-pyrene-1,3,6-trisulfonate (HPTS) and a lipophilic tetraalkylammonium base). The cross-linked polymeric beads stained with a phosphorescent iridium(III) complex are also dispersed in ethylcellulose and serve both for oxygen sensing and as a reference for HPTS. The second (pH/temperature) dually sensing system relies on the use of a pH-sensitive lipophilic seminaphthorhodafluor derivative and luminescent chromium(III)-activated yttrium aluminum borate particles (simultaneously acting as a temperature probe and as a reference for the pH indicator) which are embedded in polyurethane hydrogel layer. A silicone layer is used to spatially separate both dually sensing systems and to insure permeation selectivity for the CO2/O2 layer. The CO2/O2 and the pH/temperature layers are excitable with a blue and a red LED, respectively, and the emissions are isolated with help of optical filters. The measurements are performed at two modulation frequencies for each sensing system and the modified Dual Lifetime Referencing method is used to access the analytical information. The feasibility of the simultaneous four-parameter sensing is demonstrated. However, the practical applicability of the material may be compromised by its high complexity and by the performance of individual indicators

Methods and techniques to measure molecular oxygen in plants

Designing and developing sensors for molecular oxygen (O2) has turned into a large, interdisciplinary field of research, with significant progress seen in the past decades. Until the early 1980s, the field of O2 sensing was dominated by polarographic electrode sensors, among which the most popular Clark-type electrode found wide application in plant science. Nevertheless, the great demand for more sophisticated, intracellularly applicable O2 sensors for real-time measurements in plants cannot be satisfied by the predominant techniques. Thus, optical sensors applying an O2-specific reduction of luminescent probes or dyes provide novel, promising tools and open new perspectives on the cellular or even subcellular level of O2 measurements. This chapter aims to give a comprehensive overview on the variety of methods and systems available in the field of O2 sensing with respect to application in plant tissue. Different types of the earlier polarographic electrode technique as well as emerging alternatives will be discussed, including fluorescent proteins as potential, genetically encoded intracellular O2 sensors. Due to the tremendous variety of materials and formats, the young field of optical O2 sensing will receive particular attention directing the focus towards the progress that has been made in developing new probes and dyes. Moreover, the current state of fluorescence measurements will be explored, particularly novel, plant-specific measurement modalities that mask plant autofluorescence. For the potential user, important practical aspects are also presented, revealing the limitations of the existing methods and further encouraging more interdisciplinary research in O2 sensing