Paper-based microfluidics are microfluidic devices that consist of a series of hydrophilic cellulose or nitrocellulose fibers that transport fluid from an inlet through the porous medium to a desired outlet or region of the device, by means of capillary action. This technology builds on the conventional lateral flow test which is capable of detecting many infectious agents and chemical contaminants. The main advantage of this is that it is largely a passively controlled device unlike more complex microfluidic devices. Development of paper-based microfluidic devices began in the early 21st century to meet a need for inexpensive and portable medical diagnostic systems.
Architecture Paper-based microfluidic devices feature the following regions: Inlet: a substrate (typically cellulose) where liquids are dispensed manually. Channels: hydrophilic sub-millimeter networks that guide liquid throughout a device. Flow amplifiers: regions of varying geometry where the flow velocity is modified to impart a steady state flow of controllable velocity Flow resistors: a capillary element used to impart a reduced flow velocity in order to control the residence time of a fluid in a microfluidic deviceCapillary Flow Elements iMechanica Barriers: hydrophobic regions that prevent fluid from leaving the channel. Outlets: location where a chemical or biochemical reaction takes place.
Flow The movement of fluid through a porous medium such as paper is governed by permeability (earth sciences), geometry and evaporation effects. Collectively these factors results in evaporation limited capillary penetration that can be tuned by controlling porosity and device geometry. Paper is a porous medium in which fluid is transported primarily by wicking and evaporation. The capillary flow during wetting can be approximated by Washburn's equation, which is derived from Jurin's law and the Hagen–Poiseuille equation. The average velocity of fluid flow is generalized as, where is the surface tension, the contact angle, is the viscosity, and is the distance traveled by the liquid. More extensive models account for paper tortuosity, pore radius, and paper deformation.
Once the medium is fully wetted, subsequent flow is laminar and follows Darcy's law. The average velocity of fluid flow is generalized as, where is the medium permeability and is the pressure gradient. One consequence of laminar flow is that mixing is difficult and based solely on diffusion, which is slower in porous systems.
Manufacturing Paper-based microfluidic devices can be manufactured based on the dimensions, i.e. 2D and 3D. To fabricate 2D paper-based microfluidics, variations of methods, such as wax printing, inkjet printing, photolithography, flexographic printing, plasma treatment, laser treatment, etching (microfabrication), screen printing, digital light processing (DLP) 3-D printer, and wax screening, have been employed. Further lamination of multiple paper microfluidics creates pseudo-3D microfluidics that could provide an additional dimension of the fluidic network and increase the complexity. Each technique aims to create hydrophobic physical barriers on hydrophilic paper that passively transport aqueous solutions. Biological and chemical reagents must then be deposited selectively along the device by either dipping the substrate into a reagent solution or locally spotting a reagent onto the substrate.
Wax printing Wax printing uses a simple printer to pattern wax on paper in a desired design. The wax is then melted with a hotplate to create channels. This technique is fast and low cost, but has relatively low resolution due to the isotropy of the melted wax.
Inkjet printing Inkjet printing requires coating paper in a hydrophobic polymer, and then selectively placing an ink that etches the polymer to reveal paper. This technique is low cost with high resolution, but is limited by the speed of placing one ink droplet at a time.
Photolithography Photolithographic techniques are similar to inkjet printing, using a photomask to selectively etch a photoresist polymer. This technique has high resolution and is quick, but has high equipment and material costs.
DLP printing This technique utilizes a DLP printing technique in which photo-curable resin polymers are exposed to lights to form hydrophobic boundaries of open microchannels in a porous paper. If the effects of evaporation are of concern in the specific application then two additional layers of the curable resin can be used on the top and bottom of the channel. Excess uncured resin is then cleaned off using ethanol.Park, C., Han, Y. D., Kim, H. V., Lee, J., Yoon, H. C., & Park, S. (2018). Double-sided 3D printing on paper towards mass production of three-dimensional paper-based microfluidic analytical devices (3D-μPADs). Lab on a Chip, 18(11), 1533-1538. doi:10.1039/C8LC00367J This technique has relatively low equipment costs and utilizes readily available materials making it a promising candidate for mass production of point of care diagnostic devices.
Plasma processing In this technique, paper is first rendered hydrophobic using a hydrophobizing agent such as AKD or fluorocarbon plasma polymerization, and then O2 plasma etching with a mask is used to create hydrophilic patterns in the paper. One benefit of plasma based processes is that the complex designs and functionalities such as fully and semi-enclsoed channels,Raj N, Breedveld V, Hess D (2019) Fabrication of fully enclosed paper microfluidic devices using plasma deposition and etching Lab chip 19:3337-3343 on-off flow switches,Li X, Tian J, Nguyen T, Shen W (2008) Paper-based microfluidic devices by plasma treatment Anal Chem 80:9131-9134 and fluid flow control channelsRaj N, Breedveld V, Hess DW (2020) Flow Control in Fully Enclosed Microfluidics Paper Based Analytical Devices using Plasma Processes Sens Actuators B Chem 320:128606 can be incorporated relatively easily. However, cost of production is relatively higher than other fabrication methods.
Analytical applications
Mass spectrometry Paper-spray ionization is being rapidly developed as an interface for micro paper-based analytical devices μPAD and mass spectrometry. The technique, first described by Graham Cooks group at Purdue, involves applying a voltage to a triangular sheet of wet paper near the inlet of a mass spectrometer. Although the exact mechanism is not well understood, two modes of operation can occur: a multicone spray at high flow rates, and a single cone spray that occurs when solvent has been depleted. This is part of a larger effort to combine complex microfluidic manipulations with mass spectral detection. Wax printing hydrophobic barriers is a common method for creating distinct flow channels within paper devices, and this has been extended to μPAD-MS to enhance ionization efficiency (by enabling focusing of the analyte stream) and enable reaction mixing by wax printing on the triangular paper surface. Chromatographic separations have also been demonstrated on μPADs prior to paper-spray detection. Initially, paper-spray ionization was applied for the detection of small molecules, such as pharmaceuticals and drugs of abuse. However, it has also been shown that paper-spray ionization can ionize large proteins while retaining non-covalent interactions.
Separation methods Few analytical detectors are truly specific for a single species; therefore some type of separation step is often necessary prior to detection. Moreover, separation allows for detection of multiple analytes within a single platform. Separations based upon planar chromatography (TLC) are perhaps the easiest to implement, since many μPADs are constructed with chromatographic paper. Typically, the separation channel is defined by wax-printing two hydrophobic barriers. Electrochemical detection is perhaps most common, likely due to its ease of implementation, although colorimetry, chemiluminscence, and mass spectral detection have also been used in conjunction with paper-based chromatographic separations. Despite the ease of implementation, planar chromatography is hindered by relatively low plate height (i.e., poor separation efficiency). Since the Chakraborty group demonstrated the feasibility of electrokinetic flow on μPADs, several applications of electrophoretic separations on μPADs have appeared in the literature. The Crooks group at UT-Austin successfully demonstrated that electrophoretic separations on μPADs could be accomplished at relatively low applied voltages compared to conventional electrophoretic devices due to the high field strengths that can be generated on very thin (180 μm) sheets of origami paper. Simpler separation methods can also be used on μPADs, for instance, the Henry group demonstrated the separation of plasma from whole blood using blood separation membranes.
Flow control
There are various ways to control the fluid flow in the channels. They include changing the channel width and length, altering the wettability of the paper, diverting some fluid through a parallel channel, or changing the viscosity of the fluid. The flow in PADs can be turned off with dissolvable sugar bridges, Corona discharge treatment to alter a coating on the paper from a hydrophobic to hydrophilic state, or the use of an expandable polymer triggered by the flow to close the flow path.
Electronic integration
Integration of microfluidic platforms and electronic components have the potential to generate micro total analysis systems (μTAS), which are devices that include and automate all essential steps for sample preparation and analysis. Paper electronics rely on functional structures like conductors to be fabricated on the surface of paper, but paper-based microfluidics rely on channels and barriers to be fabricated inside the substrate. This incompatibility led to a majority of μTAS being developed using external electrodes contacted with the paper channels. However, in 2009, screen-printed electrodes were integrated into a paper-based microfluidic device to create a biosensor for glucose, lactate, and uric acid. This first report of electronic integration for paper-based microfluidics illustrated how this can improve the design of μTAS at a low cost. Since then, a variety of electrode fabrication techniques have been developed, including screen printing, inkjet printing, metal sputter deposition, pencil drawing, laser-induced pyrolysis and external electrodes to create a network of conductive traces.
Screen printing Screen printing is the most popular method to create electronic traces onto paper. In this process, the ink is transferred onto areas of the paper-based microfluidic channels using a stencil. Dungchai et al. demonstrated electrochemical sensing using screen-printed carbon ink for the working and counter electrodes and silver/silver chloride ink as the reference electrode at the end of the microfluidic channel. Screen-printed electrodes on paper-based microfluidic devices have been used not only to develop biosensors for metabolites, but also to detect bacteria and heavy metals in food and water. The scalabile nature of this process make it promising to create electrochemical devices at ultra-low cost suitable for field testing.
Inkjet printing