Fabrication of calibration plate for total internal reflection fluorescence microscopy using roll-type liquid transfer imprint lithography

摘要

A strong demand exists for the visualization of the fluid flow in microchannels, such as those in labs-on-a-chip. Multilayer nanoparticle image velocimetry (MnPIV) is a promising technique for this purpose, wherein the x and y-positions of a nanoparticle are measured using a camera and its z-position is estimated via its fluorescence intensity. To accurately estimate the z-position, however, MnPIV requires a fluorescence intensity calibration process. Water is typically used in the microfluidic devices and a nanoscale level with a refractive index matching method is conventionally used for the calibration of MnPIV. Therefore, a calibration plate with nanoscale levels whose refractive index is matched to water is necessary to calibrate the z-position of fluorescent particles in water. In this study, a fabrication technique of the calibration plate was performed using ultraviolet (UV) nanoimprint with a UV-curable resin, whose refractive index is matched to water and which possesses a high viscosity of 2900 cps. Although the residual layer thickness (RLT) of the level must be less than 100 nm to perform MnPIV, it is challenging to reduce the RLT with such a high viscosity of the resin using conventional UV nanoimprint lithography. To reduce the RLT, therefore, in this study roll-type liquid-transfer imprint lithography (LTIL) was used with the high-viscosity UV-curable resin, where a two-fold repetition of the roll-type LTIL process demonstrated a calibration pattern RLT of 40 nm or less. As a result, calibration of the MnPIV system was achieved with the calibration plate developed herein and fabricated by UV nanoimprint lithography. (C) 2017 Elsevier B.V. All rights reserved.