| Summary: | We demonstrate a method to discriminate the vertical spacing of fluorescent molecules in a guided mode structure. A function that depends only on depth and wavelength is constructed to detect the axial position of a pair of fluorescent molecules, which is especially suitable for the measurement of microstructure gaps and is not affected by the diffraction limit. Since directional fluorescence radiation information can be collected from the back focal plane in a leakage radiation microscope, we can utilize the appropriate emission angles and corresponding waveguide modes. The relative position of the fluorescent molecules is calculated by analyzing the intensity ratio between modes emission, so as to determine the vertical distance between the molecules.
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