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Extended depth of focus metal-induced energy transfer imaging with multiple axially shifted images

Because the axial resolution of an optical microscope is in the order of the wavelength of light, imaging a nano-scale object is very challenging. Many nanometer-sectioning imaging technics such as total internal reflection fluorescence (TIRF) microscopy and metal-induced energy transfer (MIET) imaging have been invented to overcome this limitation. However, the measurement ranges of these methods along the axial direction are too short to cover even a single cell. Here we propose a new long-measurement-range nanometer-sectioning imaging scheme by using MIET, focal plane shifting, and sophisticated signal analysis. We have verified the principle and the feasibility of our proposed method by using an artificial sample; it is shown that the axial measurement range is extended from 100 nm to 500 nm.