Team:Pasteur Paris/Experiments
PROTOCOLS
Microfluidics: general protocols
PDMS (Polydimethylsiloxane) is a widely used polymer in microfluidics, for its biocompatibility and transparence, among other qualities. Here we show how to prepare PDMS for microfluidic chips, as well as how to demold them, bond them to other surfaces and treat them for neuron growth. Also, we explain how our molds and chips were fabricated.
PDMS Chips Fabrication
PDMS Chip Demolding
PDMS Chip Bonding
PDMS Chip Treatment for Nerve Growth
Materials
- Sylgard 184 Elastomer Kit (Sigma-aldrich, 761036-5EA)
- Vacuum pump unit (Vacuubrand PC 3 RZ 2.5)
- Stove (Memmert UM 400)
Protocol
According to manufacturer's instruction.
- Mix monomer and curing agent (10:1 proportion) for 30 seconds
- Use a vacuum pump unit and a vacuum bell jar to extract air bubbles until the mixture is clear
- Pour mixture onto mold
- Put mixture+mold in stove at 70 degrees Celsius for 3 hours
Get full protocol here
Materials
- Razor blade (OEMTOOLS 25181 Razor Blades, 100 Pack)
- Biopsy puncher (Kai Biopsy Punch 4mm )
Protocol
- Cut the borders of the chip with the razor blade
- Extract the chip from its mold
- Drill input and output holes with the biopsy puncher
Get full protocol here
Materials
- Plasma cleaner (Diener Pico PCCE)
- Distilled water (Fisherbrand, CAS number 7732-18-5)
- Isopropanol (Fisherbrand, CAS number 67-63-0)
- Office duct tape
- Vertical laminar airflow cabinets (Euroclone aura vertical S.D.4)
Protocol
- Take chip and the surface it needs to be bonded to into the airflow cabinet
- Clean chip with duct tape and isopropanol
- Put the chip and the surface into the plasma cleaner.
- Expose chip and surface 30 seconds to plasma.
- Take the chip and the surface back in the airflow cabinet
- Press the microfluidic chip against the surface
- Insert distilled water into chip circuitry
Get full protocol here
Materials
- Poly-D-Lysine solution 1.0 mg/mL (Sigma aldrich, A-003-E)
- Laminin (Sigma aldrich, Laminin from Engelbreth-Holm-Swarm murine sarcoma basement membrane, L2020-1MG)
Protocol
- Pour poly-D-lysine with concentration 10 &mu g/mL into the chip
- Incubate over night
- Pour laminine with concentration 4 &mu g/mL
- Incubate for a few hours
Get full protocol here
Microfluidics: well chip
The well chip was designed and assembled by our team. It was used to test the biocompatibility of our membranes, as well as the culture of bacteria in the presence of current. Here we show how the molds were made, how the chip itself was assembled, how well's conductivity was measured and how bio�lm culture was performed on it.
PDMS Well Chip Mold Fabrication
Molds were made of aluminium according to the following plans (Figure 1). Part 1 Mold's center cylinder part is detachable from the bottom to make the demolding ot PDMS easier.
Microfluidics: microchannel chip
We used the microchannel chip to test the effect of NGF on the neuron's growth.
PDMS Microchannel Chip Mold Fabrication
We were allowed to use the molds made by Institut Curie. We were not involved in the process of their fabrication. Here is a short video we made about how these molds were created.