Microfluidics, a technology based on enclosed, micrometerdimension channels, first became popular in the early 1990s [1,2] as a tool for miniaturizing chemical analyses. Recently, a new form of "digital microfluidics" (DMF) has emerged, in which droplets of liquid are manipulated on an array of electrodes by means of electrowetting [3,4] and/or dielectrophoresis. [5][6][7][8] There is currently much enthusiasm for DMF, [9] as the device geometry seems a perfect match for array-based biochemical applications, such as enzyme assays [10,11] and profiling proteomics. [12][13][14] Despite this enthusiasm, the arduous procedure required to fabricate DMF devices, which typically requires metal deposition, photolithography, wet-etching, deposition or thermal growth of a dielectric layer, and deposition of a hydrophobic coating, [3,4,[11][12][13][14][15][16][17][18][19] is a barrier to its growth and development. Consequently, DMF devices are limited to use in a few laboratories worldwide.The availability of an accessible microfabrication technique is critical for the growth and continued development of DMF. An analogy can be made to the state of conventional, channel-based microfluidics in the mid-1990s. A database search reveals that approximately eight articles per year were published on the subject of channel microfluidics in 1992-1998; this number exploded to approximately 53 articles per year in 1999-2000.[20] This jump in popularity, which has greatly expanded the scope and range of applications for channel microfluidics, was driven in part by increased accessibility resulting from rapid prototyping microfabrication methods. [21][22][23] We report here, for the first time, two rapid prototyping techniques for the fabrication of DMF chips, making use of commercially available printed circuit board (PCB) substrates. In the first method, actuation electrodes were patterned on PCB substrates by using photolithography, in a manner similar to what has been reported for other applications. [24,25] This method is fast, inexpensive, and easy relative to conventional microfabrication. In the second technique, actuation electrodes were patterned directly onto substrates using a desktop laser printer; this method enabled ultra-highthroughput fabrication. We anticipate that these methods will increase the accessibility of DMF, an effect that will significantly expand the impact of this promising technology. The first rapid prototyping method, relying on photolithography, was used to form devices from two kinds of substrates: industrial-grade flexible sheets (9 lm thick copper on 50 lm polyimide) and low-grade inflexible boards (35 lm thick copper). Prior to use, devices were coated with Parylene-C (see Experimental section) or poly(dimethyl siloxane) (PDMS) as a dielectric layer, and then coated with Teflon-AF. Devices formed in this manner were used to actuate droplets sandwiched between two plates, as depicted in Figure 1a. This configuration is the most popular, as it is capable of performing all of the critical fluidic oper...