Chips and Tips

Luis G. Rigat-Brugarolas^1,2, Antoni Homs-Corbera^1,2 and Josep Samitier^1,2,3

¹ Nanobioengineering group, Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain ² Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Zaragoza, Spain.

³ Department of Electronics, Barcelona University (UB), Martí I Franques, 1, Barcelona, 08028, Spain.

Why is this useful?

At present, normal photolithographic techniques constitutes binary image transfer methodologies, where the developed pattern consists of regions with or without photoresist depending whether the UV Light has been in contact with the sample or not during the exposure process.¹ Complex 3D patterns construction is of increasing importance in the miniaturization of fluidic devices.² In the following work we introduce a photoresist-based technique to produce three-dimensional ramped microstructures for lab-on-a-chip applications.

We present a new technique that can be used to form multilevel features in SU-8 or any other negative photoresist using a single photolitograpy step, thus minimizing stages in the fabrication process in a simple and cheap way. This method thereby allows using a normal photomask without needing to add a complementary grayscale pattern, enabling complex microchannel structures.

What do I need?

• Common items and devices used in photolitographic processes (mask aligner, hot plates, chemical baths, negative photoresist and transparent substrate)
• Step Variable Metallic Neutral Density Filters (Thorlabs, Inc., NJ, USA).

What do I do?

1. Dispose the sample in the mask aligner with the SU-8 photoresist on the bottom side as depicted in Figure 1. This will force UV light to cross through the transparent substrate and to first polymerize those photoresist regions in contact with the substrate.
2. Place the photomask in the aligner standard position, normal to the light beam.
3. Select the filter (continuous, step, etc.) according to your needs (see an example in Figure 2).
4. Place the filter in the position between the photomask and the UV light source, with the filter’s design in contact with the photomask (see Figure 3).
5. Enter a correct UV exposure time; since another element is going to be added in the UV light trajectory, this value has to be adjusted (final result in Figure 4).

Fig 1: Scheme of disposal of the SU-8 photoresist in the mask aligner for achieving relief structures.

Fig 2: Example of rectangular step filter available at Thorlabs, Inc.

Fig 3: Step filter placed between the photomask and the UV light source.

Fig 4: Example of three-dimensional ramped structure constructed using SU-8. Relief characterization obtained with a profilometer. The white line represents the structure obtained after the development process (with an angle value of 30º), showing a slope from 0 µm to 12 µm (in this case, a rectangular step filter was used). The red line shows what it would look like the profile if no filter had been dispose between the photomask and the UV light source.

What else should I know?

As with any negative photoresist, grayscale exposure in conventional processes will lead to hardening the surface, removing the substrate if unattached during the development, in a methodology normally used to create cantilever structures. This is why it is important, when trying to create relief structures, to turn the sample and expose it from the glass substrate side leaving the SU-8 or any other negative photoresist on the bottom side.

Acknowledgments

We thank David Izquierdo and Juan Pablo Agusil for their technical help and for providing the material.

Reference

[1] S.D. Minteer. Microfluidic techniques: reviews and protocols. Humana Press, 2006. ISSN: 1064-3745.

[2] C. Chen, D. Hirdes, A. Folch. Gray-scale photolithography using microfluidic photomasks. PNAS, 2003. DOI:10.1073

Luis G. Rigat-Brugarolas^1,2, Antoni Homs-Corbera^1,2 and Josep Samitier^1,2,3

¹ Nanobioengineering group, Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain

² Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Zaragoza, Spain.

³ Department of Electronics, Barcelona University (UB), Martí I Franques, 1, Barcelona, 08028, Spain.

Why is this useful?

Nowadays it is common to fabricate multi-layered microfluidic microdevices by means of photolithographic techniques to create sophisticated structures allowing novel functionalities.^1,2 Without any doubt, one of the critical steps in this manufacturing process is the alignment of the different transparent layers to perform the final device.

Several approaches have been made and studied to obtain a correct structuring of the three-dimensional device like, for example, the use of gold deposition, by means of sputtering techniques, for drawing the alignment marks in the substrate,³ or using expensive mask aligners with integrated alignment protocols. Those methods are expensive and laborious. In this work we present a novel, simple and non-time-consuming methodology for drawing alignment marks using Ordyl SY330 negative photofilm, a photoresist that can be easily displayed in the substrate and, thanks to its green color, it can be readily seen in a standard microscope.

What do I need?

• Common items and devices used in photolithographic processes (mask aligner, hot plates and transparent substrate)
• Sheets of Ordyl SY330 negative photofilm.
• Ordyl Developer, SU-8 Developer or acetone.
• Photomask with the alignment marks details.
• Hot laminator.

What do I do?

A scheme of the alignment marks’ fabrication process can be seen in Figure 1. The steps are as follows:

1. Dispose the Ordyl photofim over the substrate (Figures 1A-B and 2).
2. Introduce both the substrate and the Ordyl film in a hot laminator in order to attach it firmly (see Figure 3).
3. For the exposure, use an acetate or chrome-on-glass photomask (an example can be seen in Figure 4) with the design of the alignment marks. Because Ordyl is a negative photoresist, the design should have the marks in transparent in order to polymerize them and draw them in the substrate (Figure 1C).
4. After exposure to UV Light (Figures 1D-E), an Ordyl Developer is needed (or, failing this, SU-8 developer or acetone), for having the final marks drawn in the substrate (an example can be seen in Figure 5).

Fig 1: Scheme of the fabrication process of the Ordyl alignment marks.

Fig 2: Placement of Ordyl photofilm on a microscope slide.

Fig 3: The hot laminator is used to attach the Ordyl film to the substrate.

Fig 4: Photomask with the design of the alignment marks.

Fig 5: Example of Ordyl alignment marks on a glass substrate.

Reference

[1] Dongeun Huh, Hyun Jung Kim, Jacob P Fraser, Daniel E Shea, Mohammed Khan, Anthony Bahinski, Geraldine A Hamilton and Donald E Ingber. Microfabrication of human organs-on-chips. Nature protocols. 2013 Nov. 11, vol.8. Doi:10.1038/nprot.2013.137.

[2] Michael P Cuchiara, Alicia CB Allen, Theodore M Chen, Jordan S Miller, Jennifer L West. Multilayer microfluidic PEGDA hydrogels. Biomaterials. 2010 31 5491e5497

[3] Eugene JH Wee, Sakandar Rauf, Kevin MS Koo, Muhammad JA Shiddiky, and Matt Trau. µ-eLCR: A Microfabricated Device for Electrochemical Detection of DNA Base Changes in Breast Cancer Cell Lines. LabChip. 2013 Nov 21;13(22):4385-91. DOI: 10.1039/c3lc50528f.

Vivek Kamat, KM Paknikar and Dhananjay Bodas*
Centre for Nanobioscience, Agharkar Research Institute, GG Agarkar road, Pune 411 004
E-mail: dsbodas[at]aripune.org; Tel: +91-20-25653680      

Why is this useful?       

At present many techniques are employed for fabricating channels and chambers, most of them using photolithography and soft lithography [1]. The fabrication of a circular channel and chamber in a monolithic design is challenging, which can be achieved using copper wires of varying diameters (from 20 µm). This simplistic process also eliminates usage of expensive equipment, can be performed in a normal laboratory environment (doesn’t require clean room facilities) and high fidelity structures could be obtained.      

Fabrication of chambers can be achieved in a simple, fast and novel approach by utilizing agarose gel. Agarose gel is an important component used in molecular biology experiments. Agarose powder is mixed with water and is boiled, after cooling the liquid polymerizes to form a gel. This gel can be utilized to mold the desired chamber (variable size and shape) which can be utilized for making chambers on chip.      

What do I need?      

1. PDMS (1 part curing agent and 10 part of base)
2. Agarose (1% in distilled water)
3. Copper wires of desired diameter.
4. Square box 5 x 5cm which serves as chip caster.
5. Used syringes (φ 4 mm in the present case)  

What do I do?      

1. PDMS is prepared by mixing 1:10 proportion (curing agent to base) and degassing for 30 min in a vacuum dessicator [2, 3].
2. 1% agarose powder is mixed in distilled water and boiled in microwave for 1 min until a clear solution is obtained. Decreasing the amount of agarose will result in softer gel
3. Cut the tip off of a 4 mm diameter 1 ml syringe. Pour in the agarose solution.
4. Allow the solution to cool inside the syringe and push the plunger to obtain gel in cylindrical form (see Fig 1). This cylinder so obtained can be cut into desired heights as per design requirement. In our case we have used 5 mm high cylinder for fabrication of the chamber (see Fig 2).
5. Micro dimensional copper wire is inserted through the cylinder (see Fig 3) and the whole assembly is placed in a box for molding PDMS (see Fig4) and cured at 70°C for 3 h in a convection oven.
6. After curing, place the chip in IPA for 5 min for removing the copper wire. Agarose gel can be removed by placing the chip in boiling water for 10 min. or by passing hot water using the microchannel. Repeat the process until agarose is washed completely without any traces.
7. Thus, what we have achieved is a microchannel and chamber connected together fabricated in a single step (see Figs 5 and 6). This monolith design could be extended for multiple applications such as mixing, as a reaction chamber for carrying nanoparticle synthesis, cell lysis, DNA amplification etc. [3]

Fig 1: After cooling push the plunger to get a cylindrical agarose gel

Fig 2: Cut desired height to get small cylindrical gels

Fig 3: Insert copper wire of desired diameter through the gel

Fig 4: Place in a caster box, add PDMS and allow for curing 70°C for 3 h

Fig 5: Top view of the fabricated chip

Fig 6: Fluid inside a monolithically fabricated microchannel and a chamber

References  

1. SKY. Tang and GM. Whitesides, Basic microfluidic and soft lithographic techniques in Optofluidics: Fundamentals, Devices and Applications, McGraw-Hill Professional, 2010.
2. J Friend and L Yeo, Biomicrofluidics, 2010, 4(2), 26502. DOI 10.1063/1.3259624.
3. S Agrawal, A Morarka, D Bodas and KM Paknikar, Appl Biochem Biotechnol. 2012, 167(6), 1668-77. DOI: 10.1007/s12010-012-9597-8.