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CO2-laser micromachining and back-end processing for rapid production of PMMA-based microfluidic systems
Henning Klank,* Jrg P. Kutter and Oliver Geschke Mikroelektronik Centret, DTU Building 345 East, rsteds Plads, DK-2800 Kgs. Lyngby, Denmark. E-mail: hk@mic.dtu.dk Received 2nd July 2002, Accepted 29th August 2002 First published as an Advance Article on the web 17th September 2002

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In this article, we focus on the enormous potential of a CO2-laser system for rapidly producing polymer microfluidic structures. The dependence was assessed of the depth and width of laser-cut channels on the laser beam power and on the number of passes of the beam along the same channel. In the experiments the laser beam power was varied between 0 and 40 W and the passes were varied in the range of 1 to 7 times. Typical channel depths were between 100 and 300 mm, while the channels were typically 250 mm wide. The narrowest produced channel was 85 mm wide. Several bonding methods for microstructured PMMA [poly(methyl methacrylate)] parts were investigated, such as solvent-assisted glueing, melting, laminating and surface activation using a plasma asher. A solvent-assisted thermal bonding method proved to be the most time-efficient one. Using laser micromachining together with bonding, a three-layer polymer microstructure with included optical fibers was fabricated within two days. The use of CO2-laser systems to produce microfluidic systems has not been published before. These systems provide a cost effective alternative to UV-laser systems and they are especially useful in microfluidic prototyping due to the very short cycle time of production.

Introduction
Microfluidic systems are used for various kinds of bio/chemical analyses, such as waste water monitoring, and for medical applications, such as drug delivery and point-of-care diagnostics.1 These systems have so far been mostly made using conventional silicon technology.2 As a material, silicon is a good choice when small numbers of long-lasting systems have to be produced. However, especially in clinical applications, single-use systems are desired in order to avoid contamination. Here, polymers are a good alternative to silicon, since microfluidic systems can be produced rapidly and inexpensively from polymers. For either material choice care has to be taken that the working fluids do not attack the microfluidic system. In the case of PMMA, the polymer used in this article, organic solvents cannot be used as working fluids in the system. Polymer microfluidic systems can be produced in several ways, some of which are based on replication from a micromachined master tool, while others rely on direct machining. The replication methods include hot embossing3,4 and injection molding. Silicon can be used as a master tool for hot embossing to structure a small number of complex microfluidic systems.5 The production of higher numbers of replicas on the other hand, calls for microinjection molding. Here, a more durable tool is needed, which may be machined with a computer numerical control (CNC) mill6 or which are produced by electroplating a silicon master.7 Both fabrication methods mentioned so far are not only relatively complex but also expensive compared to laser microfabrication. In this paper we present results using a CO2laser set-up,8 which was originally designed to mark objects with logos, signatures or bar codes. This laser set-up can fabricate even a complex microstructured channel system in a few seconds. Although laser microfabrication is well known, it is only UV lasers that have been used so far to produce microfluidic systems in plastic.9,10 To our best knowledge no publications have so far been published about the use of CO2laser set-ups. 242 Lab Chip, 2002, 2, 242246

Materials and apparatus

PMMA raw material (type 99530) was received from Rhm GmbH & Co. KG, Darmstadt, Germany in various thicknesses (0.5 mm6 mm) as films and plates. A conventional laboratory oven (Memmert GmbH, Schwabach, Germany) and home-built presses were used for thermal bonding. Samples were scanned under a Leo 1550 scanning electron microscope from LEO Electron Microscopy Ltd., Cambridge, UK. Ethanol (analytical grade, 97%) was purchased from Merck KGaA, Darmstadt, Germany. Bromothymol blue sodium salt in powder form was purchased from Sigma-Aldrich Denmark A/S. For the laser micromachining, a commercially available CO2laser system from Synrad Inc., Mukilteo, WA, USA was used. A computer aided design program was available to determine the movements of the laser beam and set the number of laser beam passes over a given design, the laser beam speed and the laser output power. The laser had a maximum output power of 65 W, which could be varied linearly. The laser system was equipped with a marking head containing a field lens and two swivel-mounted mirrors. The lens was able to focus the laser beam to a 185 mm diameter spot at a focal distance of 200 mm. Given the maximum output power and minimum focal spot, the maximum irradiance of the laser system was 2.3 GW m21. The mirrors allowed the placement of the focused laser beam anywhere on a 110 by 110 mm large working area. The maximum speed of the beam was 1000 mm s21.

Laser micromachining
Ablation process UV-lasers, such as excimer lasers and frequency-doubled solid state lasers, are frequently used for realizing microstructures in polymer material.11,12,13 The ablation mechanism is a complex combination of photochemical and photothermal processes.14 This means that some chemical bonds of the substance are DOI: 10.1039/b206409j

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broken directly during the process of photon absorption while others are broken thermally by the released heat of those excited molecules that do not break up photochemically. In contrast to the UV laser, a CO2-laser emits infrared radiation at a wavelength of 10.6 mm. This means that the CO2-laser beam always ablates the underlying material photothermally. The dimensions of any laser-ablated structure depend on the optical and mechanical set-up of the laser system on the one hand and on the material properties of the specific polymer on the other hand. The most important optical variables are the fnumber of the laser system and the wavelength of the laser beam in that a large f-number and a small wavelength will cause a small feature size. With respect to the material properties, the thermal diffusivity in particular should be as low as possible to ensure a small feature size. UV-lasers are typically operated in pulsed mode with pulses of nanosecond duration. These lasers have the advantage of being able to generate smaller features in comparison to a CO2laser, due to their smaller wavelength and their ablation mechanism. Either the chemical bonds are broken photochemically without creating excessive amounts of heat or the bonds are broken photothermally, in which case only small volumes are affected due to the very short laser pulses. However, the same pulses cause an uneven ablation of the treated surface15 and UV-laser equipment is relatively costly. Unlike a UV-laser, a CO2-laser emits radiation continuously. Wherever the focused laser beam meets the workpiece surface, the temperature of the irradiated spot will rise so rapidly, that the material will first melt and then decompose, leaving a void in the workpiece.16 The actual decomposition mechanism depends heavily on the strength of the chemical bonds of the monomers that make up the polymer and the structure of the polymer itself.17 In the case of PMMA, the material vaporizes in the form of monomers when it reaches its boiling temperature. This leads to cavities in the workpiece that are not contaminated by degradation components. When the laser beam is moving across the workpiece surface, it creates a pool of molten polymer in a spot where it impinges on the surface. The pool is driven away from the hot spot into all directions by the heated gases of the vaporizing plastic, while most of the melted material is resolidifying in the wake of the beam. In this manner the moving laser beam is able to cut structures such as channels and wells into the workpiece. A schematic diagram of the micromachining process can be seen in Fig. (1). The cross section of the channels depends on the thermal diffusivity of the workpiece material, i.e. on the speed of the heat dispersion in the material, and on the intensity distribution within the laser beam. Since the thermal diffusivity in polymeric materials is very low, it is the intensity distribution that mainly determines the channel cross section. An example of such a Gaussian-like profile is depicted in Fig. (2). Since the feature size of laser-cut structures depends so heavily on the material properties of the chosen polymer, especially the thermal diffusivity as well as the decomposition mechanism, we examined several different polymeric materials such as polyethyletherketone (PEEK), cyclic olefin copolymer (COC), polycarbonate (PC) and polydimethylmethoxysilane (PDMS) for their laser machinability. We found that by far the best results were achieved with PMMA, therefore we focus on PMMA in this article. In a future study, the remaining plastic materials will be investigated in greater detail. Using PMMA as a material has the additional advantage that the laser machining results can easily be compared with other micromachining techniques. It is important to note that even for a single polymer

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type such as PMMA, the material properties can differ considerably depending on the specific grade of PMMA and the manufacturer. Examples of how for instance the Vicat softening temperature alone varies from grade to grade are listed in Table 1. Apart from the choice of material, several variables are accessible to the user. The most important ones among these variables are the laser power setting, the moving speed of the laser beam and the repetition rate of the beam over the same channel. In addition, the distance between the workpiece surface and the working lens of the laser system can also be varied. If the lens-surface distance is equal to the focal length of the laser system, then the minimum spot size of the laser beam will be at the surface of the workpiece. It is important to keep in mind, that the beam will diverge beyond the minimum spot size in both directions, upwards and downwards. Therefore the channel profile may be changed within certain limits by adjusting the lenssurface distance. While keeping all other variables constant, a linear dependence between laser power and depth of the cut channels was found. As illustrated in Fig. (3), a slope of ~ 2.8 mm W21 laser power was measured for PMMA at a laser scanning speed of 381 mm s21. The cross section dimensions were measured on a fractured sample using a scanning electron microscope. Fig. (4)

Fig. 1 Schematic diagram of the laser beam, the micromachined groove, including the molten PMMA and the vaporized hemisperical cloud of MMA leaving the cut-zone.

Fig. 2 SEM micrograph of a channel cross-section in PMMA with overlayed Gaussian fit. The velocity of the laser beam was 381 mm s21.

Table 1 Vicat-softening temperature of various PMMA-grades, manufactured by Rhm GmbH, Germany Type (Manufacturers nomenclature) Vicat-softening temperature/C 6N 96 7M 104 7N 103 8N 108 7H 103 8G 113 8H 108 ZK50 75

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shows the dependence of structure dimensions on the number of laser passes at a given laser power of 6.5 W and a scan speed of 381 mm s21. The channel width increases only slightly from 240 mm to 270 mm during the first four passes and then stays constant after that, whereas the channel depth increases with a linear slope of 42 mm per pass. The narrowest channel that could be structured under these conditions was measured to be 85 mm. Depending on the material, the structures can be very accurate and smooth. However, a small risk remains that the cutting process leaves small bumps of resolidified material on the edges of the structures. Our experience shows that this happens, when channels are cut too slowly at a given intensity. At lower speeds, the gas pressure of the evaporating PMMA has more time to eject melted material out of the cutting groove. The

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slower the laser beam moves, the more melted material is ejected rather than slowly resolidifying. At a laser power setting of 52 W, the beam speed should therefore be higher than 300 mm s21, while at a power setting of 6.5 W the speed should be higher than 80 mm s21 to prevent undue large amounts of resolidified material in the channels. Fig. (5) shows a SEM-micrograph of a cross section of a PMMA channel with resolidified material at the channel side wall. The condensed polymer forms small droplets with diameters in the order of 15 mm. They can be found mainly in the upper area of the channel side walls and on top of the substrate surface, where they form a protruding bulge of excess material. We have not yet systematically studied the roughness of the machined channels. However, judging from Fig. (5) the smoothness of the wall surface can be estimated to be 1 to 2 mm, if the resolidified protrusions are ignored. The presence of the bumps as well as their form and size are specific to the material and can be an obstacle for the subsequent bonding process if their height differs a lot within the structure. If, on the contrary, the height differences are small, the bulge may even act as a built-in seal. In this case the structured surface merely has to be pressed against the hard surface of the lid. The bulge is easily deformed, since the excess polymer is usually softer than the substrate. No chemical technique has been tried yet to remove the soft excess polymer. However, we encountered no difficulties in bonding PMMA-channels having bumps of up to 30 mm as shown in Fig. (5). Back-end process/bonding

Fig. 3 Calibration curve for laser machined channels in PMMA. The channel depth increases linearly with the set laser power.The power is given in percent of 65 W. The velocity of the laser beam was 381 mm s21.

Fig. 4 Calibration curve for laser machined channels in PMMA. The channel depth increases linearly with the number of passes. The channel width changes comparatively little. Channel width and depth are defined as in Fig. (1).

Microfluidic structures such as channels and mixers often need to be diced, drilled and covered with a lid. Additionally, connections to the environment have to be made prior to use. Conventional mechanical drilling and sawing leads to rough edges and produces high material stress. In contrast, a CO2-laser system can fabricate holes and cuts with a smooth finish. It is possible to cut with high speed and high precision even through thick materials (30 mm or more). Additionally, the laser system as discussed in this article can produce throughholes in 1.5 mm thick PMMA sheets with the smallest diameter (waist) of the hole being as low as 185 mm. The throughholes are conical in shape, being broadest at the entrance and exit hole. The waist corresponds to the smallest focal spot of the laser system field lens, which has a focal length of 200 mm. Another field lens that can be mounted alternatively has a focal length of 80 mm and can produce waists of about 100 mm. Since the laser-machined holes have a conical shape, tubing can easily be inserted and fastened with a droplet of conventional epoxy glue after the bonding process. Covers for microfluidic structures should preferably, but not necessarily, be made out of the same material as used for the structure itself. If the microfluidic chip as a whole does consist

Fig. 5 SEM close-up of a cross section of a laser micromachined channel in PMMA-substrate (left picture: channel edge, right picture: close-up).

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of only one material, its thermal, chemical and electrical properties are uniform. This has several advantages, one of which is the avoidance of stress due to thermal expansion. Another important advantage is the fact that the electro-osmotic mobility is more homogeneous in such a microsystem. We investigated various methods of bonding, such as solventassisted glueing, melting, laminating and surface activation using a plasma asher. Prior to all bonding processes, it is recommended to anneal the polymer parts at a temperature slightly below the glass transition temperature of the respective polymer for at least 90 min in order to avoid stress cracks in the structure.18,19,20 Since the glass temperature of PMMA is about 105 C, we chose an annealing temperature of 80 C. Promising bonding results were achieved by lamination with commercially available lamination foil (Dansk Indbindings Kompagni), intended originally for a hot laminating machine (GBC 3500 Pro). The structure was heated up to about 100 C and the cold lamination foil was manually pressed onto the surface. Due to the low heat conductivity of the air-filled channels, the lamination foil was heated only in those regions where it touched the top layer surface. In this way, the foil adhered to the top layer surface without the lamination foil glue dissolving and clogging the channels. Hot-embossed squareshaped channels with a side length of down to 50 mm have been sealed and tested successfully with this method. As a second bonding method, we investigated surface treatment by plasma activation.21 Most polymers can be activated in an oxygen plasma and then bonded.22 Although the activation mechanism is not fully understood yet, it is likely that there is more than one process that is important for the subsequent bonding. As the most probable activation process, it is assumed that the surface is oxidized and, due to the high energy input from the plasma, radicals are formed. Possibly these radicals recombine and induce a polymerization process. The polarity of the polymer surface changes significantly after a plasma treatment, which can be seen for instance in the change of the wetting angle. However, in our experience it is not sufficient to merely press together the plasma-treated polymer surfaces without simultaneously raising the temperature. We found that reasonably strong bonds were only formed, when the surfaces were pressed together at temperatures slightly below their glass transition temperature. Another important process during plasma activation is the fact that the plasma removes dust and other foreign materials from the polymer surface and thereby cleans and polishes the substrate. Spierings et al.23 showed, that flat polymer parts can be bonded easily after polishing them thoroughly. A third mechanism that is likely to play a role is, that the polymer backbone is partly destroyed in the surface layer. If a considerable number of long-chained surface layer polymers are broken into shorter chains, this would mean that the melting temperature of the surface layer would be lower than that of the bulk material. The overall effect would be a coalescence of the surfaces while bonding, presumably leading to an adequately strong bond after cooling. In conclusion, the results of the plasma activated bonding experiments were satisfactory. While it was possible to bond a number of different polymers by surface activation in a plasma, the solvent-assisted bonding method described next was found to be faster and easier for PMMA. In order to obtain a tight bond, the surfaces were first cleaned with ethanol and then both parts, lid and structure, were immersed in ethanol for 10 min. Afterwards, the ethanol was wiped off with a lint-free tissue in order to avoid particle contamination on the surface. Any liquid ethanol in the structure has to be avoided, since channels could otherwise be clogged with softened excess material. Following the ethanol treatment, the parts were mounted in a press, which was placed in a preheated laboratory oven for 90 min at 85 C. Finally, the bonded structure was left to cool down to room temperature while remaining inside the press to avoid stress cracks in the

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structure. Fig. (6) shows a SEM-micrograph of the cross section of an ethanol-bonded PMMA-channel. The laser system together with the above described thermal bonding technique is capable of rapidly producing microfluidic systems. As an example we produced a microfluidic system, which at a later stage should be able to measure ammonia with the help of a colorimetric reaction.24,25 To test the functionality of the optical detection part of the system we used the dye bromothymol blue. This dye absorbs light mainly at around 630 nm, the same wavelength where indophenol blue, the end product of the above mentioned colorimetric reaction has its absorption maximum. Thus, bromothymol blue can be used to mimic the photometric detection of ammonia. Optical fibers were used in the microsystem to connect to the absorption cuvette within the system. We designed the layout for the optical fiber channels to form a T-shaped structure with different depths as depicted in Fig. (7). The leg of the T is narrower than the rest and hosts the optical fiber. The rest of the channel structure is deeper in order to reduce light scattering and to enable the use of index matching fluids. While laser machined structures would typically be produced on a single layer, it was also possible to fabricate multi-layer systems. The ammonia measuring microsystem presented here was built from three structured layers and a lid, all of which were bonded simultaneously. The resulting microsystem is shown in Fig. (8). In order to align the different layers, the individual layers were held by metal pins stuck in guiding holes that were cut into each layer. Optical fibers were inserted into designated channels from the edge of the structure and then glued into place. Although the optical fiber grooves are

Fig. 6 SEM micrograph of a cross section of a bonded laser micromachined channel in PMMA-substrate

Fig. 7 Schematic drawing of the cross section of the absorption cell. The T-grooves host optical fibers (narrow channel section) and can be filled with index matching fluid to increase sensitivity.

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designed to be used with index matching oil, no oil was used in the preliminary measurements presented here. Bromothymol blue solutions were inserted into the microsystem by applying droplets of the solution on top of the inlets and drawing them in by creating a vacuum on the waste outlet. The light source in the measurement set-up was a red diode laser and the detector used was a silicon photodiode detector system (HP8153A with 81520A optical head from Hewlett Packard). Measurements with a 2 mm optical pathlength cuvette showed the expected linear dependence of the absorbance on the bromothymol blue concentration between 5 mM and 100 mM. Since the molar absorptivity of the complex formed in the indophenol blue method at 630 nm is approximately 20 000 M21 cm21 and the molar absorptivity of the bromothymol blue dye26 is approximately 35 800 M21 cm21, this range would correspond to an ammonia concentration of 2.8 mM to 56.2 mM.
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this system for ammonia determination in a flow injection analysis set-up. More detailed investigations are underway.

Acknowledgement
Parts of this work have been carried out in the framework of the MicroChem-project, funded by the European Commission (BRITE/EURAM Project no. BE97-5134). Furthermore, the authors would like to thank Sren Utoft for his support during the experiments.

References
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Conclusion
Microfluidic systems in PMMA are comparatively easily produced by CO2-laser micromachining. The depth of microstructures can be controlled by the laser power, the writing speed and the number of laser beam passes, yet it is also strongly dependent on the material. Different bonding techniques have been investigated and an ethanol based bonding method was proven to be successful for bonding of a four-layer microsystem. The characteristic Gaussian-like cross sections of the micromachined channels allow self-alignment of inserted optical fibers following the bonding process. Therefore, polymeric microsystems can be designed as cheap and simple plug-and-play analyzers. First measurements with bromothymol blue showed the expected linear dependence of the absorbance on concentration. This indicates the possible use of
4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23

Fig. 8 Three-layered PMMA-microfluidic system for the detection of ammonia in aqueous samples. The system is supposed to mix the sample with three different reagents in sequence and then measure absorbance of a dye, which is the final product of the reaction sequence. In this paper only the optical part of the system was tested.

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