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DOI: 10.3791/63628-v
This article presents a streamlined protocol for creating high-fidelity protein micro patterns on soft hydrogels, specifically designed for studying cellular traction forces. The method simplifies the fabrication of isolated island patterns, crucial for controlling the shape of cell clusters.
We describe improvements to a standard method for measuring cellular traction forces, based on microcontact printing with a single subtractive patterning step of dot arrays of extracellular matrix proteins on soft hydrogels. This method allows for simpler and more consistent fabrication of island patterns, essential for controlling cell cluster shape.
This protocol allows for the consistent creation of high-fidelity protein micro patterns of whatever shape is desired, notably isolated islands of pattern for study of cell clusters. This technique can be used to make isolated island micro patterns of any shape and size in only one step, whereas previously making such patterns required two separate steps. Begin by mixing PDMS in the correct curing agent to base ratio as described in the manufacturer's instructions.
Incubate at room temperature and pressure for 15 minutes, then degas the mixture under vacuum for 15 minutes. Pour the PDMS into the master mold and transfer it to an incubator set to 37 degrees Celsius to cure overnight. Remove the master mold from the incubator and allow it to cool down to room temperature.
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