Robert Henderson, Nick Selock and Dr. Govind Rao*
Center for Advanced Sensor Technology, and Department of Chemical, Biochemical, and Environmental Engineering, University of Maryland, Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250, USA
Why is this useful?
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.
What do I need?
- Laser cutter or another implement to cut PMMA, such as a hacksaw or band saw
- An appropriate piece of equipment to anneal the PMMA (we use a convection toaster oven)
- 5.3mm thick acrylic sheet for the body of the PMMA microfluidic union
- A drill and drill bits
- Vise grips, or another way to hold the PMMA union in place while tapping
- A ¼-28 tap
- A few sheets of progressively finer sandpaper
- A 50-50% mixture of ethanol and water, along with an ultrasonic cleaner and lint-free wipes for cleaning purposes
- A strong solvent for acrylic, such as chloroform or methylene chloride
- A glass syringe with a blunt needle or needle blunted through filing
This connection scheme has been tested with good results when using the LT-115 and P-259 ¼-28 flat bottom HPLC nut and ferrule combination for 1/16″ OD tubing (Idex). This method was tested on microfluidic devices that were not bonded with the use of adhesives. This method is adaptable to other flat bottomed nuts as long as an appropriate tap and drill bit are used. Keep in mind that as the wall thickness of the PMMA union decreases, so does the strength of the connection between the HPLC nut and the device.
What do I do?
- Use a laser cutter to cut a circle with a diameter of 13mm out of the 5.3mm thick acrylic sheet. Make sure that you also either etch a center mark or make a small center hole using the laser cutter. If a laser cutter is not available, make a 13mm PMMA square using one of the other mentioned cutting implements. (Fig.1)
- Using a small drill bit, drill a pilot hole for the larger drill bit to follow in the center of the circle that you cut out in Step 1.
- Using the larger drill bit (a #3 or 0.213″ drill bit for the ¼-28 tap), drill the center hole in the cutout piece following the pilot hole drilled in Step 2.
- Using the ¼-28 tap and appropriate technique, tap the hole you have been creating in Steps 1-3. Do not use oil to lubricate the plastic as it is unnecessary and difficult to clean.
- After removing the plastic chips from the threads you just created, use progressively finer sandpaper to lap the surface that will make contact with your microfluidic device. Lapping the surface will ensure a very strong bond between the union and your device. We lap using a figure of eight motion to help guarantee a flat bonding surface. (Fig. 2)
- Anneal both your created microfluidic union and your device in an appropriate fashion. Annealing will relieve the thermal stress present in the union due to laser cutting and the drilling/tapping steps. (We use a convection toaster oven, set at 85ºC for 90 minutes, 50ºC for 30 minutes and then off with the oven door closed for 30 minutes. We find it useful to put a smooth metal plate below and above the piece to be annealed, to prevent uneven heating in the oven.)
- Thoroughly clean both the union and the microfluidic device to be bonded using a 50-50% mixture of ethanol and water. Clean only the union in an ultrasonic cleaner for 10 minutes. This mixture will not cause the union or the device to crack if they are both appropriately annealed beforehand and if the cleaning is performed within a few days of annealing.
- Place the union, lapped side down, onto the microfluidic device’s surface where the connection is desired. It is essential to center the union over the hole which receives the fluid or gas from the HPLC tubing. We use a small jig composed of two different sized plastic dowels to help us in this step (the black object in the bottom right corner of Fig.3).
- Pull some solvent into the glass syringe.While holding the union in place using your jig, gently drip some solvent from the glass syringe at the interface between the union and your microfluidic device (start with a single drop). The solvent will creep into the joint between the union you created and your microfluidic device due to capillary action. Add enough solvent to fill the gap between the union and your device, but be careful not to get any solvent into the fluid passages within your device.
- Being careful not to disturb the union’s position, gently apply pressure on the union until the surfaces become lightly bonded using your gloved fingers (Fig. 3). Light bond is usually achieved in less than a minute. In this step, also attempt to add enough pressure to force out any bubbles at the interface.
- Remove the alignment jig and add more solvent around the interface between the union and your device. This step reinforces the union’s connection to the microfluidic device.
- Put a small mass directly on top of the union, and leave the bonding union overnight to achieve the strongest connection between the union and your device.
- After the piece has dried overnight, remove the mass and then use the connection as you would any other flat bottom HPLC union. (Figure 4)
- Figure 1. The dimensions of the raw PMMA union
- Figure 2. Lapping the surface of the microfluidic device with a figure-eight motion
- Figure 3. Lightly bonding the union to the microfluidic device using gentle pressure
- Figure 4. The finished microfluidic union
The following YouTube video ‘Power Tools & Carpentry Skills : How to Use a Tap & Die Set’ by Expert Village helps explain the use of taps (Step 4): http://www.youtube.com/watch?v=6DDhw191MLo
The authors would like to acknowledge the work of Dr. Yordan Kostov, Mike Tolosa and Mike Frizzell.