Method for arraying biomolecules and for monitoring cell motility in real-time

1. A method for arraying biomolecules comprising the steps of:

positioning a first layer to be in fluid-tight contact with a support, the first layer having an upper surface and defining a pattern of micro-orifices, each micro-orifice of the pattern of micro-orifices having walls and defining a micro-region on the support when the first layer is placed in fluid-tight contact with the support such that the walls of said each micro-orifice and the micro-region on the support together define a micro-well;

positioning a second layer to be in fluid-tight contact with an upper surface of the first layer, the second layer defining a pattern of macro-orifices, each macro-orifice of the pattern of macro-orifices having walls and defining a macro-region when the first layer is placed in fluid-tight contact with the support and the second layer is placed in fluid-tight contact with the first layer such that the walls of the macro-orifice and the macro-region together define a macro-well; and

immobilizing at least one biomolecule of a plurality of biomolecules in each respective micro-region on the support so as to situate the at least one biomolecule within a corresponding micro-well, the biomolecules thereby being arrayed on the support in a pattern that corresponds to the pattern of the micro-orifices.

2. The method of claim 1 , wherein the step of immobilizing at least one biomolecule includes immobilizing a cell.

3. The method of claim 2 , wherein the step of immobilizing at least one biomolecule includes immobilizing a portion of at least one cell.

4. The method of claim 1 , each macro-region encompasses at least one micro-region.

5. The method of claim 4 , each macro-region encompasses a plurality of micro-regions.

6. The method of claim 1 , wherein the step of immobilizing at least one biomolecule comprises immobilizing a single type of biomolecule in each micro-region.

7. The method of claim 1 , wherein the step of immobilizing at least one biomolecule comprises immobilizing a different type of biomolecule in each micro-region.

8. The method of claim 1 , wherein the step of immobilizing at least one biomolecule includes immobilizing at least two different types of biomolecules in a single micro-region.

9. The method of claim 1 , further comprising the step of applying a coating to an upper surface of the support.

10. The method of claim 9 , wherein the step of applying a coating to an upper surface of the support includes applying to the upper surface of the support a coating that is made of a material selected from the group consisting of proteins, protein fragments, peptides, small molecules, lipid bilayers, metals and self-assembled monolayers.

11. The method of claim 1 , further comprising enclosing at least one of the macro-wells with at least one cap.

12. A method for arraying biomolecules comprising:

positioning a first layer to be in fluid-tight contact with a support, the first layer having an upper surface and defining a pattern of micro-orifices, each micro-orifice of the pattern of micro-orifices having walls and defining a micro-region on the support when the first layer is placed in fluid-tight contact with the support such that the walls of said each micro-orifice and the micro-region on the support together define a micro-well;

immobilizing at least one biomolecule of a plurality of biomolecules in each respective micro-region on the support so as to situate the at least one biomolecule within a corresponding micro-well, the biomolecules thereby being arrayed on the support in a pattern that corresponds to the pattern of the micro-orifices;

removing the first layer from the support after the step of immobilizing; and

positioning a second layer to be in fluid-tight contact with the support, the second layer defining a pattern of macro-orifices, each macro-orifice of the pattern of macro-orifices having walls and defining a macro-region when the second layer is placed in fluid-tight contact with the support such that the walls of the macro-orifice and the macro-region together define a macro-well.

13. The method of claim 12 , wherein the step of immobilizing at least one biomolecule includes immobilizing a cell.

14. The method of claim 13 , wherein the step of immobilizing at least one biomolecule includes immobilizing a portion of at least one cell.

15. The method of claim 12 , each macro-region encompasses at least one micro-region.

16. The method of claim 15 , each macro-region encompasses a plurality of micro-regions.

17. The method of claim 15 , wherein the step of immobilizing at least one biomolecule comprises immobilizing a single type of biomolecule in each micro-region.

18. The method of claim 15 , wherein the step of immobilizing at least one biomolecule comprises immobilizing a different type of biomolecule in each micro-region.

19. The method of claim 12 , wherein the step of immobilizing at least one biomolecule includes immobilizing at least two different types of biomolecules in a single micro-region.

20. The method of claim 12 , further comprising the step of applying a coating to an upper surface of the support.

21. The method of claim 20 , wherein the step of applying a coating to an upper surface of the support includes applying to the upper surface of the support a coating that is made of a material selected from the group consisting of proteins, protein fragments, peptides, small molecules, lipid bilayers, metals and self-assembled monolayers.

22. The method of claim 12 , further comprising enclosing at least one of the macro-wells with at least one cap.

23. A method for arraying biomolecules comprising:

positioning a first layer to be in fluid-tight contact with a support, the first layer having an upper surface and defining a pattern of micro-orifices, each micro-orifice of the pattern of micro-orifices having walls and defining a micro-region on the support when the first layer is placed in fluid-tight contact with the support such that the walls of said each micro-orifice and the micro-region on the support together define a micro-well;

positioning a second layer to be in fluid-tight contact with the support, the second layer comprising a plurality of rings, the rings defining a pattern of respective macro-orifices, each ring having walls and defining a macro-region when the second layer is placed in fluid-tight contact with the support such that the walls of the ring and the macro-region together define a macro-well; and

immobilizing at least one biomolecule of a plurality of biomolecules in each respective micro-region on the support so as to situate the at least one biomolecule within a corresponding micro-well, the biomolecules thereby being arrayed on the support in a pattern that corresponds to the pattern of the micro-orifices.

24. The method of claim 23 , wherein the step of immobilizing at least one biomolecule includes immobilizing a cell.

25. The method of claim 23 , wherein the step of immobilizing at least one biomolecule includes immobilizing a portion of at least one cell.

26. The method of claim 23 , each macro-region encompasses at least one micro-region.

27. The method of claim 26 , each macro-region encompasses a plurality of micro-regions.

28. The method of claim 23 , wherein the step of immobilizing at least one biomolecule comprises immobilizing a single type of biomolecule in each micro-region.

29. The method of claim 23 , wherein the step of immobilizing at least one biomolecule comprises immobilizing a different type of biomolecule in each micro-region.

30. The method of claim 23 , wherein the step of immobilizing at least one biomolecule includes immobilizing at least two different types of biomolecules in a single micro-region.

31. The method of claim 23 , further comprising the step of applying a coating to an upper surface of the support.

32. The method of claim 31 , wherein the step of applying a coating to an upper surface of the support includes applying to the upper surface of the support a coating that is made of a material selected from the group consisting of proteins, protein fragments, peptides, small molecules, lipid bilayers, metals and self-assembled monolayers.

33. A method for arraying biomolecules comprising:

positioning a layer to be in fluid-tight contact with the support, the layer defining a pattern of macro-orifices, each macro-orifice of the pattern of macro-orifices having walls and defining a macro-region when the layer is placed in fluid-tight contact with the support such that the walls of the macro-orifice and the macro-region together define a macro-well;

inserting each plug of a set of plugs in a respective macro-well, each of the plugs comprising a lower membrane placed in fluid-tight contact with the support when the layer is placed in fluid-tight contact with the support and the plug is received in a corresponding macro-well defined by the layer and the support, the lower membrane further defining a pattern of micro-orifices, wherein each micro-orifice has walls and defines a micro-region on the support when the plug is in fluid-tight contact with the support such that the walls of the micro-orifice and the micro-region together define a micro-well; and

immobilizing a biomolecule in at least one micro-region on the support so as to be situated within the micro-well, such that the biomolecule is arrayed on the support in a pattern that corresponds to the first pattern of micro-orifices.

34. The method of claim 31 , further comprising enclosing at least one of the macro-wells with at least one cap.