Abstract
Total internal reflection fluorescence microscopy (TIRFM) is a powerful tool for analyzing 3D fluid flow in microchannel. To visualize the 3D fluid flow via TIRFM, intensity calibration of a fluorescent particle is required by using calibration plates, carrying a set of nano-steps of different heights. In the present study, to discern the micro-structures in water, TIRFM visualization is carried out by a refractive index-matching method using a water employed as a working fluid. The MEXFLON obstacles is chosen to be able to adjust its refractive index to match to that of the water with an index of 1.333. We have demonstrated a fabrication technique for making the micro-structures on a glass substrate by employing thermal nanoimprint. In summary, we have succeeded in the fabrication of micro-structures made of a polymer resin and used it for the visualization of the fluorescence intensity in a water.
