Team:BIOTEC Dresden/Methods Vesicles
From 2009.igem.org
Vesicles - Methods
Setup of the microfluidic system
The microfluidic system consists of a flow chamber made of Polydimethylsiloxane (PDMS) and a pump system that controls the flow rates of the various liquids into the chamber. Droplets are created within a defined space in the chamber and are propagated along a grid that allows containment and imaging. Two types of chambers have been used, differing in the geometry of the space where droplets were produced. One featured T-junction, and the other a V-junction (Fig1).
Fig1: Different geometries of the intersection between aqueous and oil phase in flow chambers
Production of flow chambers:
- mix PDMS and curing agent in 10:1 ratio
- degas and pour on wafer with etched microstructures
- polymerize on heat plate at 150 ºC for 30 min
- add unpolymerized PDMS mixture to points on microstructure where microtube inlets are to be pierced
- polymerize on heat plate at 150 ºC for 30 minutes
- remove polymerized PDMS from wafer, cut to fit onto glass cover slide (24 x 60 mm), and use clean needles (0.8 mm) or laser cutter to pierce tube inlets
- ionize PDMS and glass slide in plasma chamber for 30 sec to make it reactive
- align PDMS on glass slide and seal
- seal irreversibly by heating on plate at 60ºC for 6 hours