Multiwell insert device that enables label free detection of cells and other objects

• US 20040091397A1
• Filed: 11/07/2002
• Published: 05/13/2004
• Est. Priority Date: 11/07/2002
• Status: Abandoned Application

• Current Assignee: Corning Incorporated
• Original Assignee: Corning Incorporated
• Inventors: Picard, Laurent A.G.
• Application Number:
  US10/290,001
• Publication Number:
  US 20040091397A1
• Time in Patent Office: Days
• US Class Current: 422/400
• CPC Class Codes: B01D 67/0088 Physical treatment with compounds, e.g. swelling, coating or impregnation
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 3/50255 Multi-well filtration
• CPC Class Codes: B01L 3/50853 with covers or lids
• CPC Class Codes: C12M 25/04 in combination with well or multiwell plates, i.e. culture inserts
• CPC Class Codes: G01N 21/41 Refractivity; Phase-affecting properties, e.g. optical path length G01N21/21 takes precedence

Abstract

A multiwell insert device and method for using the multiwell insert device are described herein. The multiwell insert device includes an upper chamber, a lower chamber, a membrane and a sensor for detecting in a label-free manner an object (e.g., cells, molecules, proteins, drugs, chemical compounds, nucleic acids, peptides, carbohydrates) that passed through the membrane from the upper chamber into the lower chamber by measuring a change in a refractive index caused by the object being present on a surface of the lower chamber. The multiwell insert device can be used to perform a wide-variety of assays including, for example, cell migration assays and drug permeability assays. In addition, the multiwell insert device can form or be incorporated into a well of a microplate.

Claims

1. A multiwell insert device, comprising:

an upper chamber;

a lower chamber;

a membrane located between said upper chamber and said lower chamber; and

a sensor for detecting an object that passed through said membrane from said upper chamber into said lower chamber by measuring a change in a refractive index caused by the object being present on a surface of said lower chamber.

2. The multiwell insert device of claim 1, wherein said object is a cell, molecule, protein, drug, chemical compound, nucleic acid, peptide or carbohydrate.

3. The multiwell insert device of claim 1, wherein said sensor is used to assess the migration capacity of the objects through said membrane.

4. The multiwell insert device of claim 1, wherein said sensor is used to assess the attachment capacity of the object on said membrane.

5. The multiwell insert device of claim 1, wherein said membrane is a polyester membrane or a polycarbonate membrane.

6. The multiwell insert device of claim 1, wherein said membrane is a microporous membrane with pores in the range of 0.1 μ

m to 12.0 μ

m.

7. The multiwell insert device of claim 1, wherein said membrane is coated with a biochemical component, protein, biological membrane or cells.

8. The multiwell insert device of claim 1, wherein said surface of said lower chamber is coated with at least one substance that favors the capture and concentration of said object at the surface of said lower chamber.

9. The multiwell insert device of claim 1, wherein said surface of said lower chamber is coated with a capture reagent including an antibody or other binding proteins.

10. The multiwell insert device of claim 1, wherein said surface of said lower chamber is coated with a capture reagent including hydrophobic, hydrophilic or charged surface chemistries.

11. The multiwell insert device of claim 1, wherein said sensor is a grating-based planar waveguide sensor.

12. The multiwell insert device of claim 11, wherein said grating-based planar waveguide sensor utilizes an angular interrogation approach to detect the object.

13. The multiwell insert device of claim 11, wherein said grating-based planar waveguide sensor utilizes a spectral interrogation approach to detect the object.

14. The multiwell insert device of claim 1, wherein said sensor is a grating-based surface plasmon resonance sensor.

15. The multiwell insert device of claim 1, wherein said sensor is a prism-based surface plasmon resonance sensor.

16. The multiwell insert device of claim 1, wherein a plurality of said multiwell insert devices form a plurality of wells in a microplate.

17. The multiwell insert device of claim 1, wherein said lower chamber is shallow compared to said upper chamber.

18. The multiwell insert device of claim 1, wherein said sensor contacts said membrane.

19. A method for using a multiwell insert device, said method comprising the steps of:

placing an object in an upper chamber of said multiwell insert device;

allowing the object to migrate from the upper chamber to a lower chamber through a membrane of said multiwell insert device; and

detecting the object on a surface of the lower chamber using a sensor that measures a change in a refractive index caused by the object being present on a surface of the lower chamber.

20. The method of claim 19, wherein said object is a cell, molecule, protein, drug, chemical compound, nucleic acid, peptide or carbohydrate.

21. The method of claim 19, wherein said sensor is used to assess the migration capacity of the object through said membrane.

22. The method of claim 19, wherein said sensor is used to access the attachment capacity of the object on said membrane.

23. The method of claim 19, wherein said membrane is a polyester membrane or a polycarbonate membrane.

24. The method of claim 19, wherein said membrane is a microporous membrane with pores in the range of 0.1 μ

m to 12.0 μ

m.

25. The method of claim 19, wherein said membrane is coated with a biochemical component, protein, biological membrane or cells.

26. The method of claim 19, wherein said surface of the lower chamber is coated with at least one substance that favors the capture and concentration of said object at a surface of the lower chamber.

27. The method of claim 19, wherein said surface of said lower chamber is coated with a capture reagent including an antibody or other binding proteins.

28. The method of claim 19, wherein said surface of said lower chamber is coated with a capture reagent including hydrophobic, hydrophilic or charged surface chemistries.

29. The method of claim 19, wherein said sensor is a grating-based planar waveguide sensor.

30. The method of claim 29, wherein said grating-based planar waveguide sensor utilizes an angular interrogation approach to detect the object.

31. The method of claim 29, wherein said grating-based planar waveguide sensor utilizes a spectral interrogation approach to detect the object.

32. The method of claim 19, wherein said sensor is a grating-based surface plasmon resonance sensor.

33. The method of claim 19, wherein said sensor is a prism-based surface plasmon resonance sensor.

34. The method of claim 19, wherein a plurality of said multiwell insert devices form a plurality of wells in a microplate.

35. The method of claim 19, wherein said lower chamber is shallow compared to said upper chamber.

36. The method of claim 19, wherein said sensor contacts said membrane.

37. A microplate, comprising:

a frame including a plurality of wells formed therein, each well is in the form of a multiwell insert device that includes;

an upper chamber;

a lower chamber;

a membrane located between said upper chamber and said lower chamber; and

a sensor for detecting an object that passed through said membrane from said upper chamber into said lower chamber by measuring a change in a refractive index caused by the object being present on a surface of said lower chamber.

38. The microplate of claim 37, wherein said object is a cell, molecule, protein, drug, chemical compound, nucleic acid, peptide or carbohydrate.

39. The microplate of claim 37, wherein said sensor is used to assess migration capacity of the object through said membrane.

40. The microplate of claim 37, wherein said sensor is used to access the attachment capacity of the object on said membrane.

41. The microplate of claim 37, wherein said sensor is used to perform multiplex assays.

42. The microplate of claim 37, wherein said sensor is a grating-based planar waveguide sensor.

43. The microplate of claim 37, wherein said sensor is a grating-based surface plasmon resonance sensor.

44. The microplate of claim 37, wherein said sensor is a prism-based surface plasmon resonance sensor.