Improving Sample Distribution Homogeneity in Three-Dimensional Microfluidic Paper-Based Analytical Devices by Rational Device Design Giorgio Gianini Morbioli Thiago Mazzu-Nascimento Luis Aparecido Milan Amanda M. Stockton Emanuel Carrilho 10.1021/acs.analchem.6b04953.s003 https://acs.figshare.com/articles/media/Improving_Sample_Distribution_Homogeneity_in_Three-Dimensional_Microfluidic_Paper-Based_Analytical_Devices_by_Rational_Device_Design/4891769 Paper-based devices are a portable, user-friendly, and affordable technology that is one of the best analytical tools for inexpensive diagnostic devices. Three-dimensional microfluidic paper-based analytical devices (3D-μPADs) are an evolution of single layer devices and they permit effective sample dispersion, individual layer treatment, and multiplex analytical assays. Here, we present the rational design of a wax-printed 3D-μPAD that enables more homogeneous permeation of fluids along the cellulose matrix than other existing designs in the literature. Moreover, we show the importance of the rational design of channels on these devices using glucose oxidase, peroxidase, and 2,2′-azino-bis­(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) reactions. We present an alternative method for layer stacking using a magnetic apparatus, which facilitates fluidic dispersion and improves the reproducibility of tests performed on 3D-μPADs. We also provide the optimized designs for printing, facilitating further studies using 3D-μPADs. 2017-04-12 00:00:00 wax-printed 3 D -μPAD ABTS optimized designs layer devices glucose oxidase Three-Dimensional Microfluidic Paper-Based Analytical Devices alternative method Rational Device Design Paper-based devices cellulose matrix Sample Distribution Homogeneity 3 D -μPADs sample dispersion Three-dimensional microfluidic paper-based fluidic dispersion layer treatment