To begin open the plate map software and select the culture cells in the 3D option. Then, select imaging model for 96 well plates. In the matrix selection window, select HydroGel PX-2.53.
Enter the cell types as glutamatergic neurons and astrocytes, and cell density as 20 million cells per milliliter. Design the desired plate map by highlighting wells on the plate. Include at least one row of 2D control on plastic in the design.
Then, verify that the plate model listed at the top of the window is changed to the intended plate model for use. Using the Download buttons, download the protocol and print file for the plate map. Then, save them to the bio printer-linked computer.
Prepare a 0.1%polyethylenimine solution in a borate buffer and filter it through a 0.22-micron filter. Add 100 microliters of 0.1%polyethylenimine solution to each 2D control well of a 96-well plate, and incubate at 37 degrees Celsius for two hours. Then, rinse the wells five times with PBS.
Once the wells are dried, add diluted laminin solution into the wells and incubate at 37 degrees Celsius for four hours.
