Automated addressable microfluidic technology for minimally disruptive manipulation of cells and fluids within living cultures

A microfluidic plumbing system comprising:

• an array of addressable microfluidic ports that include a plurality of inlet ports and that serve as access openings that allow fluid flow to and from different locations associated with a cell culture chamber;
• an inflatable on-chip valve layer comprising inflatable on-chip valves, wherein the inflatable on-chip valves are addressable and are able to be actuated to open or closed states, which allow or block fluid flow to and from individual addressable microfluidic ports from among the array of addressable microfluidic ports;
• one or more flexible membrane layers disposed adjacent the inflatable on-chip valve layer;
• one or more fluid flow layers disposed adjacent the one or more flexible membrane layers, wherein the array of addressable microfluidic ports are disposed within the one or more fluid flow layers and wherein each addressable microfluidic port defines a path for fluid flow;
• and
• a cell culture chamber that is not a multi microwell plate and that is located adjacent the one or more fluid flow layers and in fluid communication with the array of addressable microfluidic ports;
• wherein the cell culture chamber defines a single culturing chamber or scaffold pore space in fluid communication with at least two of the plurality of inlet ports included in the array of addressable microfluidic ports;
• and
• wherein the array of addressable microfluidic ports deliver fluid flow to or from one or more selected locations within the single culturing chamber or scaffold pore space.