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Ratiometric Imaging with Oxygen PEBBLEs
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Oxygen PEBBLEs have been used for ratiometric fluorescence imaging in C6 glioma cells. This figure shows a Nomarski (differential phase contrast) image (top - a) of the cultured cells on a coverslip. The next set of images are the green and red channels (b,c) of a confocal image showing the separate fluorescence from the reference dye and the oxygen sensing dye. Computer software has then be used (d,e) to superimpose the images, showing where signals are coming from within the cells. Changes in the red fluorescence can be used to show local changes in oxygen concentration, and ratio images can be generated showing oxygen concentration based on test-tube calibrations of the sensors. |
