Three-Dimensional, Paper-Based Microfluidic Devices Containing Internal Timers for Running Time-Based Diagnostic Assays

This chapter describes a method for fabricating three-dimensional (3D), paper-based microfluidic devices that contain internal timers for running quantitative, time-based assays. The method involves patterning microfluidic channels into paper, and cutting double-sided adhesive tape into defined patterns. Patterned paper and tape are assembled layer by layer to create 3D microfluidic devices that are capable of distributing microliter volumes of a sample into multiple regions on a device for conducting multiple assays simultaneously. Paraffin wax is incorporated into defined regions within the device to provide control over the distribution rate of a sample, and food coloring is included in defined regions within the device to provide an unambiguous readout when the sample has reached the bottom of the device (this latter feature is the endpoint of the timer).