Thin layer fluorescence microscopy based on one-dimensional photonic crystal

Abstract

A new method of specimen illumination for wide-field fluorescence microscopy has been presented. This method allows to excite the fluorescence in a thin near-surface layer of the studied object. As a result, the captured images have greater contrast and signal-to-background ratio in comparison with the epifluorescence ones. The long-range surface waves in one-dimensional photonic crystal have been used to localize the electromagnetic field exciting the fluorescence. An experimental setup has been created to excite the surface waves and obtain images of the objects from the near-surface layer. For an illustration of the possibilities of our method, we conducted several experiments with specimens that are typical for fluorescence microscopy, such as bacteria and eukaryotic cells.