How to detach glass coverslips from acrylamide gel?

Detach the patterned glass coverslip from the acrylamide gel using a scalpel, make sure that the gel is fully immersed during the entire detachment process, otherwise you will end up with collapsed micropatterns. Rinse the acrylamide gel attached to silanized coverslip (acrylamide coverslips) in PBS several times.

Are polyacrylamide hydrogels biologically relevant?

In conclusion, we presented a bench-ready method to fabricate polyacrylamide hydrogels of biologically relevant stiffness with topographical features of biologically relevant sizes that can be applied to a range of cell-related studies that require an independent control of the stiffness and the geometry of the environment.

How to polymerize glass coverslips?

Add TEMED to the acrylamide solution, mix briefly but vigorously. Add APS solution to the acrylamide solution, mix briefly but vigorously. Put a drop of 7 μL/cm 2 of the acrylamide polymerization mix on each glass coverslip previously placed on parafilm. Slowly place the other coverslip of interest on top while taking care to avoid bubbles.

Is a polystyrene-coated coverslip effective?

Interestingly, if a polystyrene-coated coverslip was used for the template production, the transfer was then inefficient. The polystyrene coating has been shown to increase micropatterns stability for cell lines producing strong forces.

How do you sterilize acrylamide coverslips?

Warm up your cell culture reagents as usual. Using ethanol sterilized tweezers, transfer the acrylamide coverslip in sterile tissue culture petri dish filled with sterile PBS (gel facing up). Rinse once with PBS and cover with warmed medium. In the meantime resuspend your cell in warmed medium.

Can polyacrylamide hydrogels be used for topographical structures of different shapes?

We have developed a reliable protocol to obtain topographical structures of different shapes (lines, square posts, and square holes) of biologically relevant sizes (5–10 μ m) on polyacrylamide hydrogels of biologically significant stiffnesses (3–145 kPa).