Miniaturized cell array methods and apparatus for cell-based screening

A method of making a substrate for selective cell patterning comprising:

• a) providing a substrate with a surface, wherein the surface contains hydroxyl groups;
• b) contacting the hydroxyl groups on the surface of the substrate with a bifunctional molecule comprising a hydroxy-reactive moiety and a nucleophilic moiety to form a monolayer;
• c) applying a stencil to the substrate;
• and
• d) further modifying the substrate by a technique selected from the group consisting of
• i) applying an electrophilic cell repulsive moiety to exposed regions of the monolayer to form a covalent bond between the cell repulsive moiety and the nucleophilic moiety of the bifunctional molecule deposited in step b to produce cell repulsive locations, removing the stencil, and applying cell adhesive molecules to the substrate to produce cell binding locations, wherein the cell adhesion molecules bind the substrate only in positions that were contacted by the stencil;
• and
• ii) applying cell adhesive molecules to exposed regions on the monolayer to produce cell binding locations, removing the stencil, and applying an electrophilic cell repulsive moiety to the substrate to produce cell repulsive locations, wherein the cell repulsive moiety forms a covalent bond with the nucleophilic moiety of the bifunctional molecule deposited in step b only in positions that were contacted by the stencil.