Apr 22, 2023
Version 1
Custom open-chamber microfluidic fabrication V.1
- Quyen Do1,2,3,
- Jimena Baleriola4,
- Nora Bengoa-Vergniory1,2,3,5,6,6,
- Richard Wade-Martins1,2,3
- 1Oxford Parkinson's Disease Centre and Department of Physiology, Anatomy and Genetics, University of Oxford, South Park Road, Oxford OX1 3QU, United Kingdom;
- 2Kavli Institute for Neuroscience Discovery, University of Oxford, Dorothy Crowfoot Hodgkin Building, South Park Road, Oxford OX1 3QU, United Kingdom;
- 3Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, MD, 20815, USA;
- 4Achucarro Basque Center for Neuroscience, Leioa, Spain University of the Basque Country (UPV/EHU), Department of Neuroscience, Leioa, Spain;
- 5Achucarro Basque Center for Neuroscience, Leioa, Spain;
- 6Ikerbasque - Basque Foundation for Science, Bilbao, Spain
Protocol Citation: Quyen Do, Jimena Baleriola, Nora Bengoa-Vergniory, Richard Wade-Martins 2023. Custom open-chamber microfluidic fabrication. protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvr47q2gmk/v1Version created by Cláudia C. Mendes
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 22, 2023
Last Modified: April 22, 2023
Protocol Integer ID: 77446
Keywords: Open microfluidic, co-culture, microcircuit, chamber microfluidic fabrication this protocol, microfluidic fabrication, chambered microfluidic, microfluidics suitable for cell, master mould, acquired master mould, fabrication, chamber, cell
Funders Acknowledgements:
Aligning Science Across Parkinson’s
Grant ID: ASAP-020370
Abstract
This protocol described the fabrication of two- and three-open chambered microfluidics suitable for cell culturing using commercially acquired master mould.
Materials
Reagents:
- SYLGARD™ 184 Silicone Elastomer Kit (Dow, CAT# 1673921)
- Geltrex™ LDEV-Free Reduced Growth Factor Basement Membrane Matrix (ThermoFisher Scientific, CAT# A1413202)
- Poly-D-Lysine (ThermoFisher Scientific, CAT# A3890401)
Equipment:
- Custom master mould containing both duo- and trio-chambers (manufactured in Microliquid, Spain).
- Fisherbrand™ Glass Circle Coverslips (ThermoFisher Scientific, CAT# 12323138)
Before start
Always store cut-out devices in ethanol.
Avoid exposing microfluidic devices, particularly the cell-adherent side, to open air and unnecessary contact. Once sterilised, devices should be handled with care following sterile working techniques.
Casting of microfluidic devices
A custom master mould containing both duo- and trio-chambers (Figure 1) is manufactured by and obtained from Microliquid (Spain).
Figure 1: Custom master mould.
Mix 46 ml of silicon elastomer SylGARD 184, Dow Corning © with 4 mL of its curing agent (supplied in the same pack), i.e. the 1:10 ratio in a 50 ml falcon.
Mix well by inverting the falcon multiple times till a homogenous solution is seen by eye.
Slowly pour the mixture into the master mould and incubate for 2-2.5 hours at 60ºC.
Note
Varying incubation time can alter the device stiffness.
Device preparation
Manually remove the final cast from the mould using a sharp blade.
Manually cut out individual devices and immediately wash them with 100% ethanol.
Air-dry cut-out devices in tissue culture hood.
Place dry and sterile devices on ethanol – sterilised 19 mm-diameter coverslips to form an instantaneously tight seal.
Coat both chambers and microchannel area with Poly-D-Lysin (0.1 mg/ml) overnight followed by Geltrex™ prior to cell culturing.