Microelement device

• US 6,315,940 B1
• Filed: 05/13/1999
• Issued: 11/13/2001
• Est. Priority Date: 11/16/1996
• Status: Expired due to Fees

• Current Assignee: Cytocentrics AG (Cytocentrics Bioscience GmbH)
• Original Assignee: NMI Naturwissenschaftliches und Medizinisches Institut an an der Universität Tübingen. (NMI Natural and Medical Sciences Institute at the University TÜBINGEN)
• Inventors: Stelzle, Martin, Nisch, Wilfried, Stett, Alfred, Egert, Ulrich
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US09/311,780
• Time in Patent Office: 915 Days
• Field of Search: 435/283.1, 435/285.2, 435/287.1, 435/287.3, 435/288.3, 435/288.4, 435/173.1, 435/173.2, 435/173.3, 435/173.4, 435/173.5, 435/173.6, 435/173.7, 435/173.8, 435/173.9, 422/98, 422/101, 204/403, 204/600, 204/450, 204/409, 204/400, 436/63, 436/177, 436/149, 436/150, 436/151
• US Class Current: 435/287.1
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: C12M 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: G01N 21/763 Bioluminescence
• CPC Class Codes: G01N 33/48728 Investigating individual cells, e.g. by patch clamp, voltage clamp investigating individual particles in general G01N15/10

Abstract

A microelement device has a plurality of microelements, which may be configured as microelectrodes, arranged on a substrate and adapted for making contact to cells present in a liquid environment. The cells are guided onto the microelectrodes, are isolated or are mechanically attracted to the microelectrodes. A negative-pressure force or a hydrodynamic force may be applied on the cells. Also described are a method for making contact to the cells, and a method for manufacturing the microelement device is disclosed.

Claims

1. A microelement device comprising:

a substrate having a plurality of cavities for receiving a liquid containing biological cells;

a plurality of microelements each of which is arranged at a bottom of one of said cavities, each said microelement having a contact surface facing upwardly for electrically contacting one of said biological cells;

a plurality of channels, each extending within said substrate and being in fluid communication with one of said microelements; and

a negative pressure source in fluid communication with each said channel for attracting said cells toward said microelements.

2. The device as defined in claim 1, wherein said microelements are connected to amplifier means and wherein a reference electrode is provided for contacting a liquid received with said cavities.

3. The device as defined in claim 2, wherein the microelectrode comprises a surface being in contact with the environment and a surface being in contact with the cell, said surface being in contact with the environment being larger than the surface being in contact with the cell.

4. The device as defined in claim 1, wherein said microelements are connected to a voltage source, and wherein a reference electrode is provided for contacting a liquid received within said cavities.

5. The device as defined in claim 1, wherein said negative pressure source includes a pump means for said liquid.

6. The device as defined in claim 1, wherein said cavities are configured as funnel-like microcuvettes, said microelements being located at the bottom of said microcuvettes.

7. The device as defined in claim 6, further comprising surface regions between said microcuvettes, said regions being coated with a cell-repelling material.

8. The device as defined in claim 6, wherein said microcuvettes are coated with a cell-attracting layer of coating material.

9. The device as defined in claim 6, wherein said substrate comprises at least a base plate and a cover plate located thereabove.

10. The device as defined in claim 9, wherein the base plate is made of a material selected from the group consisting of glass, quartz, silicon, or plastic.

11. The device as defined in claim 9, wherein the cover plate is made of a material selected from the group consisting of glass, quartz, silicon, plastic, polystyrene, PMMA, and polyimide.

12. The device as defined in claim 9, wherein at least one element selected from the group consisting of the base plate and the cover plate is made of a material that is transparent to light, the wavelength of the light lying in a region of the spectrum accessible to microscopy techniques.

13. The device as defined in claim 9, wherein the base plate is connected to the microelements and comprises an edge connector which is guided outward laterally.

16. The device as defined in claim 1, wherein the microelements comprise sensing electrodes and stimulus electrodes.

17. The device as defined in claim 16, wherein the microelements comprise reference electrodes.

18. The device as defined in claim 1, wherein the microelements comprise multiple concentrically arranged individual electrodes.

19. The device as defined in claim 6, wherein the microelements comprise at least one reference electrode and at least one sensing electrode, wherein said sensing electrode is arranged at the bottom of said microcuvette, and wherein said reference electrode is arranged at a distance above said sensing electrode.

14. The device as defined in 9, wherein the base plate comprises at least a lower signal processing plate and an element plate located thereabove.

15. The device as defined in claim 14, wherein the signal processing plate is guided outward laterally as an edge connector.

20. A microelement device comprising

a substrate having a plurality of cavities for receiving a liquid containing biological cells;

a plurality of electrical microelements each of which is arranged at a bottom of one of said cavities, each said microelement having a contact surface facing upwardly for electrically contacting one of said biological cells and being electrically connected to a voltage source establishing an electric field directed for electrostatically attracting one of said cells toward said microelement, and

a reference electrode for contacting liquid contained in said cavities.

21. The device as defined in claim 20, wherein said cavities are configured as funnel-like microcuvettes, said microelements being located at the bottom of said microcuvettes.

22. The device as defined in claim 21, further comprising surface regions between said microcuvettes, said regions being coated with a cell-repelling substrate.

23. The device as defined in claim 21, wherein said microcuvettes are coated with a cell-attracting substrate.

24. The device as defined in claim 21, wherein said substrate comprises at least a base plate and a cover plate located thereabove.

25. The device as defined in claim 21, wherein the microelements comprise at least one reference electrode and at least one sensing electrode, wherein said sensing electrode is arranged at the bottom of, said microcuvette, and wherein said reference electrode is arranged at a distance above said sensing electrode.

26. The device as defined in claim 20, wherein the microelements comprise sensing electrodes and stimulus electrodes.

27. The device as defined in claim 20, wherein the microelements comprise reference electrodes.

28. The device as defined in claim 20, wherein the microelements comprise multiple concentrically arranged individual electrodes.

29. A microelement device comprising

a substrate comprising a plurality of cavities for receiving a liquid containing biological cells;

a plurality of electrical microelements each of which is arranged at a bottom of one of said cavities, each said microelement having a contact surface facing upwardly for electrically contacting one of said biological cells;

a plurality of channels, each extending within said substrate and being in fluid communication with one of said cavities; and

an electroosmotic pump means cooperating with each said channel for attracting one of said cells toward one of said microelements.

30. The device as defined in claim 29, wherein said electroosmosis pump comprises two electrodes electrically coupled to opposite ends of said channels, a voltage being applied between the electrodes.

31. The device as defined in claim 29, wherein said cavities are configured as funnel-like microcuvettes, said microelements being located at the bottom of said microcuvettes.

32. The device as defined in claim 31, further comprising surface regions between said microcuvettes, said regions being coated with a cell-repelling substrate.

33. The device as defined in claim 31, wherein said microcuvettes are coated with a cell-attracting substrate.

34. The device as defined in claim 31, wherein said substrata comprises at least a base plate and a cover plate located thereabove.

35. The device as defined in claim 29, wherein the microelements comprise sensing electrodes and stimulus electrodes.

36. The device as defined in claim 29, wherein the microelements comprise reference electrodes.

37. The device as defined in claim 29, wherein the microelements comprise multiple concentrically arranged individual electrodes.

38. The device as defined in claim 31, wherein the microelements comprise at least one reference electrode and at least one sensing electrode, wherein said sensing electrode is arranged at the bottom of said microcuvette, and wherein said reference electrode is arranged at a distance above said sensing electrode.