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This work demonstrates the use of a commercially-available single-sided rubber adhesive sheet to interface a rigid microfluidic chip for fluid access. This interconnect is an alternative to PDMS-based ports, whose plasma bonding characteristics may not be applicable for all rigid materials. The adhesive component of the port is incorporated with the rubber sheet; hence no additional curing time is needed. This technique provides a quick and inexpensive method to interface rigid microfluidic chips.

When making devices that are direction-specific and very small, you need to check them in the microscope each time to see which side to start your flow. With this technique you can mark the PDMS with a color marker that does not interfere with the device. When working with a multilayer device that has multiple valves and channels it is convenient to have identification markers.

Owing to the advantages in miniaturization and cellular microenvironmental control, microfluidic devices have been increasingly applied to cell biology research [1]. Particularly, microfluidic devices can precisely configure chemical concentration gradients and flexibly manipulate the gradient conditions in space and in time [2, 3]. Various microfluidic gradient-generating devices have been used for studying cell migration and chemotaxis [2, 3]. These studies rely on live cell microscopy and usually only one experiment can be performed at a time. Previously, a double gradient device was demonstrated for parallel cell migration experiment with a motorized stage to image cells in different gradient channels [4]. However, the full XYZ motorized stage is expensive and thus often times limits the practical use of high-throughput microfluidic devices.

To overcome this limitation, here we report a stacked microfluidic device that allows parallel live cell imaging experiments on a single chip with only a Z motorized stage. This device is fabricated with multiple stacked layers of PDMS devices, and the cell imaging channels in each layer are aligned so they all fit into a single microscope viewing field. Thus, by only adjusting the vertical focus using a Z motorized stage, multiple cell channels can be imaged repeatedly over time. If a full XYZ motorized stage is available, the throughput of the stacked device can be further increased along horizontal dimensions. Making a stacked device is straightforward and this strategy can be useful for improving experiment throughput, especially in a limited microscopy facility.

[1] G. B. Salieb-Beugelaar et al., Latest developments in microfluidic cell biology and analysis systems. Anal. Chem., 2010, 82, 4848-4864.[2] S. Kim, H. J. Kim, and N. L. Jeon, Biological applications of microfluidic gradient devices. Integr. Biol., 2010, 2, 584-603.[3] J. Li and F. Lin, Microfluidic devices for studying chemotaxis and electrotaxis. Trends Cell Biol., 2011, 21, 489-497.[4] W. Saadi et al., A parallel-gradient microfluidic chamber for quantitative analysis of breast cancer cell chemotaxis. Biomed. Microdevices, 2006, 8, 109-118.

[1] Jie Xu and Daniel Attinger, How to prevent sagging during the bonding or lamination of chips with large aspect ratio chambers, Chips & Tips (Lab on a Chip), 24 July 2009.[2] D. Di Carlo, D. Irimia, R. G. Tompkins and M. Toner, Continuous inertial focusing, ordering, and separation of particles in microchannels. Proc. Natl. Acad. Sci. U. S. A., 2007, 104, 18892-18897.[2] D. Di Carlo, J. F. Edd, D. Irimia, R. G. Tompkins and M. Toner, Equilibrium separation and filtration of particles using differential inertial focusing. Anal. Chem., 2008, 80, 2204-2211.

The use of HPLC fittings and tubing in microfluidics is becoming more commonplace as the use of microfluidics in the sciences increases. Even though acrylic or PMMA is one of the most common plastic substrates used in microfluidics, the ability to strongly and easily connect an HPLC fitting to a PMMA chip has until now been elusive. Although there are some commercially available HPLC-to-chip products from Idex, they require heating the microfluidic chip well above the glass transition temperature of PMMA to create a permanent epoxy bond between attachment and chip. For these reasons, we present a simple scheme to strongly connect a standard HPLC flat bottom nut to a PMMA microfluidic device through the creation and bonding of an easy-to-make ¼-28 PMMA microfluidic union, using common lab equipment.

This methodology allows for very quick fabrication of a wide range of PDMS films using off the shelf components which are common in a traditional laboratory and office or can be easily purchased. The final goal is to fabricate films which feature different thicknesses, according to the original film used as spacer, and different hardnesses, by changing the ratio of PDMS to cross linker. The film can be stored and used as needed to cut off parts such as gaskets, spacers, etc. Here in the lab we use them to make microfluidic chambers of specific thickness.

Figure 2. Completed paper microfluidic chips made using Parafilm®. (a.) Spiral design has a channel width of 1 mm. (b.) Design has circles of 4mm diameter and 8 straight channels of 2mm width and 10mm length. (c.) Blue dye added to the paper microfluidics chip shown in b.

For heavier weight paper such as Whatman Grade 1 filter paper, it is necessary to apply higher pressure (~200 psi) such as in a hot press to produce the microchips. However, for lighter weight paper such as Kimtech Kimwipes and VWR light-duty tissue wipes, microchips can simply be made by heating the plates of a heating element such as a hair straightener, and then applying gentle pressure to the paper, PC, Parafilm® stack to produce the paper-based chips.

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