Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466, Gatersleben, Germany.
PreSens Precision Sensing GmbH, Josef-Engert-Strasse 11, 93053, Regensburg, Germany.
New Phytol. 2012 Nov;196(3):926-936. doi: 10.1111/j.1469-8137.2012.04295.x. Epub 2012 Sep 17.
Biological samples are far from homogeneous, with complex compartmentation being the norm. Major physiological processes such as respiration do not therefore occur in a uniform manner within most tissues, and it is currently not possible to image its gradients in living plant tissues. A compact fluorescence ratiometric-based device is presented here, consisting of an oxygen-sensitive foil and a USB (universal serial bus) microscope. The sensor foil is placed on the sample surface and, based on the localized change in fluorescence signal over time, information about the oxygen consumption (respiration) or evolution (photosynthesis) can be obtained. Using this imaging technique, it was possible to demonstrate the spatial pattern of oxygen production and consumption at a c. 20-μm level of resolution, and their visualization in the rhizosphere, stem and leaf, and within the developing seed. The oxygen mapping highlighted the vascular tissues as the major stem sink for oxygen. In the leaf, the level of spatial resolution was sufficient to visualize the gas exchange in individual stomata. We conclude that the novel sensor set-up can visualize gradients in oxygen-consuming and producing processes, thereby facilitating the study of the spatial dynamics of respiration and photosynthesis in heterogeneous plant tissues.
生物样本远非均匀的，复杂的隔室化是常态。因此，大多数组织中的主要生理过程（如呼吸）不会以均匀的方式发生，目前还无法在活体植物组织中对其梯度进行成像。本文提出了一种基于紧凑荧光比率的设备，它由一个氧敏感箔片和一个 USB（通用串行总线）显微镜组成。传感器箔片放置在样品表面上，根据荧光信号随时间的局部变化，可以获得关于氧气消耗（呼吸）或演化（光合作用）的信息。使用这种成像技术，可以以约 20-μm 的分辨率展示在根际、茎和叶以及发育中的种子中氧气产生和消耗的空间模式，并对其进行可视化。氧映射突出了血管组织作为氧气的主要茎汇。在叶子中，空间分辨率足以可视化单个气孔的气体交换。我们得出结论，新的传感器装置可以可视化耗氧和产氧过程的梯度，从而促进对异质植物组织中呼吸和光合作用空间动态的研究。
