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Prototyping Paper-Based Microfluidics
Joshua Pearce, N. Anzalone, C. Heldt
To cite this version:
Joshua Pearce, N. Anzalone, C. Heldt. Open-Source Wax RepRap 3-D Printer for Rapid Proto-typing Paper-Based Microfluidics. Journal of Laboratory Automation, 2016, 21 (4), pp.510-516. �10.1177/2211068215624408�. �hal-02113499�
Open-source Wax RepRap 3-D Printer for Rapid Prototyping Paper-Based Microfluidics
J. M. Pearce1,2,*, N. C. Anzalone3, and C. L. Heldt4
1. Department of Materials Science & Engineering, Michigan Technological University, MI 2. Department of Electrical & Computer Engineering, Michigan Technological University, MI 3. Department of Mechanical Engineering & Engineering Mechanics, Michigan Technological University, MI 4. Department of Chemical Engineering, Michigan Technological University, MI *Corresponding author: Michigan Technological University, 601 M&M Building, 1400 Townsend Drive, Houghton, MI 499311295 (pearce@mtu.edu) ph: 9064871466
Keywords: 3-D printing, wax, paper-based microfluidics, lab-on-a-chip, electrochemical detection
Abstract The opensource release of selfreplicating rapid prototypers (RepRaps), has created a rich opportunity for lowcost distributed digital fabrication of complex three dimensional objects such as scientific equipment. For example, 3D printable reactionware devices offer the opportunity to combine open hardware microfluidic handling with lab on a chip reactionware to radically reduce costs and increase the number and complexity of microfluidic applications. To further drive down the cost while improving the performance of labonachip paperbased microfluidic prototyping this study reports on the development of a RepRap upgrade capable of converting a Prusa Mendel RepRap into a wax 3D printer for paperbased microfluidic applications. An opensource hardware approach is used to
demonstrate a 3D printable upgrade for the 3D printer, which combines a heated syringe pump with the RepRap/Arduino 3D control. The bill of materials, designs, basic assembly and use instructions are provided along with a completely free and open source software tool chain. The open source hardware device described here accelerates the potential of the nascent field of electrochemical detection combined with paperbased microfluidics by dropping the marginal cost of prototyping to nearly zero while accelerating the turnover between paperbased microfluidic designs. Introduction Bowyer's opensource release of selfreplicating rapid prototypers (RepRaps) [13], has created a rich opportunity for lowcost distributed digital fabrication of complex three dimensional objects. There has been a surge of interest among scientists from a wide swath of fields in using RepRap 3D printers to design, manufacture and share the opensource digital designs of scientific equipment [46]. As these opensource digitally replicable scientific tools are freely distributed and thus widely accessible for the cost of materials, 3D printing components or entire devices results in substantial economic savings – generally 9099% less than proprietary commercial equipment [5]. Preliminary analysis of the economic value of an open hardware approach [7] indicates scientific funders can obtain a return on such investments in the hundreds or thousands of percent [8]. This has resulted in an explosion of highquality 3D printable lab tools for: biology labs [9] and biotechnological and chemical labwares [1012], optics and optical system components [13], colorimetery [14], nephelometer [15] and turbidimeter [16], nitrate testing [17], liquid autosampling [18] with robot arms [19], automated sensing arrays [20] and compatible components for medical applications [21]. Not only are relatively inexpensive 3D printed parts being used to replace conventional scientific tools, they are
also being used to create new scientific opportunities for investigation. For example, utilizing a 3D printer in the lab makes it easier for researchers to design and build laboratory tools faster and more closely aligned to specific research needs. For example, a highly configurable reaction vessel made with a 3D printer can integrate reagents, catalysts and a purification apparatus into a single reactionware [22,23], which can significantly simplify multistep reactions by integrating equipment into a single component. These 3D printable reactionware devices offer the opportunity to combine open hardware microfluidic handling [24] with simple “labonachip” reactionware [25].
There is a large interest in creating such inexpensive labonachip devices for rapid detection of health-related biomarkers [26,27], explosives [28,29], and gas components [30]. The goal of such
applications is to save lives by detecting molecules in short periods of time, typically less than one
hour. While the field of microdevices is currently exploding, cost is a constant concern with these
devices. In 2008, the Whitesides group pioneered paper microfluidics [31]. This discovery was quickly
followed by other complex 3-D structures made in paper that controlled the capillary flow of biological
liquids by creating channels bordered with hydrophobic wax [32-34]. Such paper microfluidic devices
have reduced or eliminated the need for pumping of fluids, and this reduces both the footprint and cost
of the overall device. The paper base is also inexpensive and readily available throughout the world,
making it an ideal point of care platform. Originally, groups were creating wax channels using
lithography [32], but this is time consuming and expensive. Recently, wax printing has been developed
that provides more flexibility in design and reduces the cost of device creation [33].
To further drive down the cost while improving the performance of lab on a chip paper microfluidic prototyping this study reports on the development of a RepRap upgrade capable of converting a Prusa Mendell RepRap into a wax 3D printer for paperbased microfluidic applications. An opensource hardware approach is used to demonstrate a 3D printable upgrade for the 3D printer,
which combines a heated syringe pump with the RepRap/Arduino 3D control. The bill of materials, designs, basic assembly and use instructions are provided along with a completely free and open source software tool chain. The results of this approach are discussed.
Materials and Methods
The mechanical and electrical design of the wax 3-D printer upgrade followed standard RepRap
design methods for scientific equipment outlined in detail elsewhere [5], thus relying on only open
source electronics, RepRap printable custom parts, off-the-shelf globally available non-printed parts
(referred to as vitamins in the RepRap literature), and a completely open source software tool chain.
The designs are released under a CC-BY-SA license. In addition, no expensive tools, specialty
equipment or advanced skills are needed for assembly.
Both the custom 3-D printed parts as well as the wax print designs are created in OpenSCAD
[35]. Open-source and freely available OpenSCAD is a script-based, parametric CAD program
possessing powerful 3-D modeling capabilities. 3-D models are created by adding and subtracting
primitive objects created by code to produce the desired complex shapes. The script language is based
upon C++. As only a small number of methods are required to produce very complex designs the
learning curve is short for students with programming experience even if they are not familiar with
conventional CAD. The scripts are written such that designs are parametric, thus by changing the
values of a single variable the design can easily be altered. Models rendered in OpenSCAD are saved
as a (.scad) file and are also exported as a stereolithography (.stl) file for the slicing process. Slicing
converts an stl file into layers of equal thickness in the z direction in G-code, a human-readable file
format specifying the path the print head must follow to produce a physical object. Cura, an open
The mass of the printed parts was calculated with Cura Lulzbot Edition 14.09-1.18 using
normal quality setting for polylactic acid (PLA) for the syringe carriage and the remainder of the parts
were printed in acrylonitrile butadiene styrene (ABS) for its relatively higher temperature resistance.
Results and Discussion
The syringe design was derived from the Wijnen et al. open-source 3-D printable syringe pump
library [37]. There are five 3-D printed custom parts needed for the conversion including a motor
mount (Figure 1), syringe carriage (Figure 2), x carriage (Figure 3) and two syringe retainers (Figure
4). The 3-D printed parts can be printed in about 5 hours on any standard RepRap 3-D printer
consuming $3.11 of commercial PLA filament. The entire bill of materials including part, count, cost
and source totals $87.49 as seen in Table 1. This BOM assumes that only a single conversion kit is
purchased and it should be pointed out that there are discounts for most of the components when
purchased in bulk (e.g. fasteners). The conversion kit assembly follows the basic steps outlined by
Wijnen et al. [37] for syringe pumps and is shown completed without the syringe in Figure 5.
The Prusa Mendell RepRap [38], which can be fabricated for less than $600 and assembled in
24 hours [39], is itself capable of printing all of the conversion components (Figures 1-4). After
printing the ABS and PLA components with the standard direct drive head, the mechanical components
of the syringe depressor are assembled as shown in Figure 5 and the heater made up of the copper pipe
and nichrome heating element wrapped in Kapton polyimide tape for the syringe is installed (Figure 6).
Then the syringe is placed into position along with the temperature feedback provided by the thermistor
and the assembly is insulated as shown in Figure 7. Finally, the extruder driver and standard hot end is
removed to be replaced by the assembled syringe pump (Figure 8). It should be noted that similar
work is needed to convert delta style 3-D printers with Bowden sheaths.
The wax printer, can then be fully assembled, utilizing a Prusa Mendel open source RepRap 3-D
printer for the frame and electronics as shown in Figure 8. The resultant wax 3-D printer has a
horizontal print area of 150 mm width by 200 mm length, and about 30 mm in height. The printer uses
an open source electronics prototyping platform of the Arduino Atmega 2560 [40], with a RepRap
Arduino Mega Pololu Shield (RAMPS) to control the motors and heating element [41]. The glass
syringe is then filled with a wax and heated to melting and the wax is printed on a paper substrate as
shown in Figure 9 at 5mm/s.
Similar to the design of the conversion kit, the design of the microfluidic device takes place in
OpenSCAD using parametric variables (e.g. changing the gap width variable alters the distance
between the two lines of wax in a test print). The design is then exported as an stl in order to be sliced
with Cura into several layers of G-code, a language that is understood and controls the movements of
the 3-D printer. Thus complex geometries for the wax print can be easily designed and printed as
shown in the example in Figure 10. The settings for Cura, that are used to dictate the boundaries and
how the motors move, are specific to each printer. With the wax printer, Cura is set up to produce slow
moving prints, around 5 mm/s, with an extrusion rate less than 0.5 mm/s for the wax (as compared to
conventional RepRap printing materials), in order to maintain print quality. The print is meant to be
printed, hot enough to melt the wax, but is close enough to the phase transition temperature to have the
wax solidify shortly after it is deposited on the print bed. In the example shown in Figures 9 and 10 the
Cerita wax (normally used for investment casting) is printed at 70oC. The G-code is then imported into
one of several printer control software packages such as Repetier [42], Printrun [43] or Franklin [44]),
which provides a user interface to control the printer. This software is used to load the G-code into the
the marginal cost of changing a paper-based microfluidic design is reduced to the cost of the substrate
paper and the wax used for the experiment. The far more time consuming and expensive
lithography-based prototyping steps, including transparency mask fabrication, SU-8 photoresisit for the channels,
wafer cleaning, lithography, developing, sealing, and vacuum degassing, are eliminated.
Conservatively, creating a single new microfluidic design using lithography based methods costs more
than the wax printer conversion kit capable of unlimited design options for nearly zero marginal cost. It
however, should be noted that there are other lower cost methods of microfluidic design available, such
as the paper-based methods in [33]. Future work is needed to compare these two methods, which would
involve first optimizing the printing of the 3-D wax printer described. This is accomplished for each
specific wax in a short power vs velocity study. First, an approximate extrusion temperature is found
by ramping up the temperature while printing a constant velocity spiral. When this is done a simple zig
zag pattern is designed that increases the print head velocity at each corner. Then this pattern is
repeatedly printed with a small x-offset while varying the extrusion temperature and extrusion rate in a
systematic fashion. After the optimal print head velocity and extrusion rate are found a final set of runs
are made while adjusting the temperature again by small increments to obtain optimal printing
parameters. These parameters are then used in printing identical microfluidic devices using the two
methods and the resultant devices are tested.
The Prusa has an x-y position accuracy of 20 microns and a minimum z accuracy of 0.4
microns. The heated bed on the Prusa can also be used to anneal the wax and/or further drive it into the
paper substrate to obtain channel widths that the printer itself is not capable of obtaining. This,
however, can only be achieved for a subset of geometries. Future work is needed to improve this
resolution, by for example moving to finer pitched lead screws and belts, higher-quality stepper motors,
and easy removal of the plunger for wax filling; multi material printing with multiple heads, and
indirect automated head swapping.
In addition, to wax printing to replicate the work that has been accomplished in the past [33] for lower costs so that experiments are more accessible to a greater selection of scientists [45], this 3D printer also enables fabrication of fully integrated microdevices using a single tool and the creation of complex 3D geometries for yet unexplored microdevice applications. For example, The most common detection method for paper microfluidic devices is colorimetric [46,47] and there are already well established techniques for utilizing Arduinobased open source electronics and RepRap 3D printing to make handheld detection devices [14,15,17]. However, colorimetric detection works well for yes/no detection, but is difficult to calibrate to a quantitative measures in microfluidic applications. For a microfluidic device this is the difference between a pregnancy test and a blood glucose level test. A more quantitative measure uses electrochemical detection. Electrochemical paperbased devices have been created to detect the oxidation of glucose with an indiumdoped tin oxide electrode [47] and a more standard glassy carbon electrode [48]. Other examples include human tears being analyzed for ion detection [49] and the detection of cholesterol levels in human serum [50]. While electrochemical detection combined with paperbased microfluidics is still in its infancy, it has a high potential to become a useful and cost effective microdevice platform for many different analytes, as well as serving to detect environmental hazards. The open source hardware device described in this paper accelerates this potential by dropping the marginal cost of prototyping microdevices to nearly zero, while accelerating the turnover between paperbased microfluidic designs. Acknowledgements
The authors would like to acknowledge NSF (CBET-1159425 and CBET-1510006) for funding of this
work.
Declaration of Conflicting Interests
The authors declared no potential conflicts of interest with respect to the research, authorship, and/or
publication of this article.
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Tables and table legends
Table 1. Bill of materials for wax 3-D printer conversion kit.
Part Name Quantity $/unit Cost $ Source
Glass Syringe, 3 ml 1
$ 9.93
$
9.93 Zoro.com
30 RW x 2" Blunt Needle 1 $ 14.00 $ 14.00 Labemco.com
LM6UU Linear Bearing 2
$ 0.65
$
1.30 Robotdigg.com
Semitec 104-GT2 Thermistor 1 $ 1.12 $ 1.12 Mouser.com
1" x 24" Copper Pipe 1
$ 4.89
$
4.89 HomeDepot.com
Nema 11 Motor 1 $ 15.50 $ 15.50 Kysanelectronics.com
5 mm x 5 mm Shaft Coupling 1
$ 2.98
$
2.98 Adafruit.com
625zz Ball Bearing 1 $ 7.77 $ 7.77 VXB.com
M5 Hex Nut 2 $ 0.17 $ 0.34 McMasterCarr.com M5 Washer 1 $ 0.14 $ 0.14 McMasterCarr.com M3 Hex Nut 8 $ 0.30 $ 2.43 McMasterCarr.com
M3 x 16 mm socket head cap screw 8 $ 0.82 $ 6.56 McMasterCarr.com
M2.5 x 12mm socket head cap screw 4
$ 0.17
$
0.68 McMasterCarr.com
Stainless Steel M5 Threaded Rod 150mm 1 $ 2.61 $ 2.61 McMasterCarr.com
6 mm A2 Tool Steel Smooth Rod 150mm 2
$ 6.83
$
13.66 McMasterCarr.com
Ni-Chrome Wire 150mm 1 $ 0.48 $ 0.48 McMasterCarr.com
insulation 1 scavenged
Vitamin subtotal $ 84.38
Printed Parts mass (g)
Motor mount (ABS) 39 $ 42.95 $ 1.68 Lulzbot.com
Syringe carriage (PLA) 18
$ 24.95
$
0.45 Lulzbot.com
Syringe retainers (ABS) 6 $ 42.95 $ 0.26 Lulzbot.com
X carriage (ABS) 17
$ 42.95
$
0.73 Lulzbot.com
Printed part subtotal $ 3.11 Total
$ 87.49
Figure Legends
Figure 1. Cura rendering of motor mount.
Figure 2. Cura rendering of syringe carriage.
Figure 3. Cura rendering of x carriage.
Figure 4. Cura rendering of the small and large syringe retainers.
Figure 5. Assembled conversion kit without syringe.
Figure 6. Assembled conversion kit with heater and copper pipe installed.
Figure 7. Fully assembled conversion kit with syringe, thermistor wiring and insulation.
Figure 8. Prusa Mendel open source RepRap Converted Wax 3-D printer.
Figure 9. Wax printing on paper substrate.
