Marine Biological Section, Department of Biology, University of Copenhagen, Strandpromenaden 5, DK-3000 Helsingør, Denmark.
Water Res. 2011 Oct 15;45(16):5027-37. doi: 10.1016/j.watres.2011.07.007. Epub 2011 Jul 13.
A ratiometric luminescence intensity imaging approach is presented, which enables spatial O2 measurements in biofilm reactors with transparent planar O2 optodes. Optodes consist of an O2 sensitive luminescent dye immobilized in a 1-10 μm thick polymeric layer on a transparent carrier, e.g. a glass window. The method is based on sequential imaging of the O2 dependent luminescence intensity, which are subsequently normalized with luminescent intensity images recorded under anoxic conditions. We present 2-dimensional O2 distribution images at the base of a tap water biofilm measured with the new ratiometric method and compare the results with O2 distribution images obtained in the same biofilm reactor with luminescence lifetime imaging. Using conventional digital cameras, such simple normalized luminescence intensity imaging can yield images of 2-dimensional O2 distributions with a high signal-to-noise ratio and spatial resolution comparable or even surpassing those obtained with expensive and complex luminescence lifetime imaging systems. The method can be applied to biofilm growth incubators allowing intermittent experimental shifts to anoxic conditions or in systems, in which the O2 concentration is depleted during incubation.
提出了一种比率型发光强度成像方法，该方法可使用透明平面氧敏光学传感器在生物膜反应器中进行空间氧测量。光学传感器由氧敏发光染料固定在透明载体（例如玻璃窗口）上的 1-10 μm 厚的聚合物层中组成。该方法基于对氧依赖性发光强度的顺序成像，随后将其与在缺氧条件下记录的发光强度图像进行归一化。我们展示了用新的比率法在自来水生物膜底部测量的二维 O2 分布图像，并将结果与在相同生物膜反应器中用荧光寿命成像获得的 O2 分布图像进行比较。使用传统的数码相机，这种简单的归一化发光强度成像可以产生具有高信噪比和空间分辨率的二维 O2 分布图像，与昂贵且复杂的荧光寿命成像系统获得的图像相当，甚至更好。该方法可应用于生物膜生长培养箱，允许在实验过程中间歇性地切换到缺氧条件，或者在培养过程中 O2 浓度耗尽的系统中使用。
