METHOD OF REDUCING CURVATURE IN A MENISCUS OF LIQUID MEDIUM

• US 20100047845A1
• Filed: 02/26/2008
• Published: 02/25/2010
• Est. Priority Date: 02/26/2007
• Status: Active Grant

• Current Assignee: Stemcell Technologies Inc. (Stemcell Technologies Canada Inc.)
• Original Assignee: Stemcell Technologies Inc. (Stemcell Technologies Canada Inc.)
• Inventors: Dossantos, Gary, Woodside, Steven, Dowd, Jason, Egeler, Oliver
• Granted Patent: US 9,062,283 B2
• Time in Patent Office: Days
• US Class Current: 435/29
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/161 Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: C08J 7/0427 with only one layer of a composition containing a polymer binder with more layers C08J7/042
• CPC Class Codes: C08J 7/043 Improving the adhesiveness of the coatings per se, e.g. forming primers adhesives in the form of films or foils characterised by the primer layers between the polymer carriers and the adhesives C09J7/50
• CPC Class Codes: C12M 23/10 Petri dish crystallising dishes B01L3/06
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/20 Material Coatings immunocoatings C12M25/00

Abstract

The present application is directed to methods of improving cell culture vessel assays. In one aspect the application is directed to a method of reducing the curvature of the meniscus comprising applying a coating material to the interior wall of the vessel, wherein the coating material provides a receding contact angle of about 90 degrees with aqueous solutions and culture media. In another aspect, the application is directed to a method of labeling cells in a first solution by generating droplets of a second solution containing at least one cell-labelling agent and allowing the droplets of the second solution to contact the surface of the first solution.

Claims

1. A method of reducing curvature in a meniscus of liquid medium in a culture vessel comprising applying a coating material to interior wall surfaces of the culture vessel, wherein the coating material provides a receding contact angle between the vessel wall and the liquid of approximately 90 degrees with aqueous solutions and culture media.

2. The method as claimed in claim 1, wherein the receding contact angle is about 75 degrees to about 110 degrees.

3. The method as claimed in claim 1, wherein the receding contact angle is about 85 degrees to about 105 degrees.

4. The method as claimed in claim 1, wherein the coating material inhibits adhesion of molecular constituents present in aqueous solution or cell culture medium, so as to prevent alteration of the surface properties of the coating.

5. The method as claimed in claim 1, wherein the vessel is made of polystyrene, polypropylene, polycarbonate, polyvinylchloride or polytetrafluoroethylene.

6. The method as claimed in claim 1, wherein the coating material is silicone based, fluoropolymer based, petroleum jelly, paraffin wax, EPDM or Buna Nitrile or is an insert that is made of silicone, EPDM or Buna Nitrile or that is coated with a coating material that is silicone based, fluoropolymer based, petroleum jelly, paraffin wax, EPDM or Buna Nitrile.

7. The method as claimed in claim 6, wherein the silicone-based material comprises non-crosslinked siloxane, methylsiloxane, methylvinylsiloxane, or copolymers thereof.

8. The method according to claim 1, wherein the vessel is a culture dish or multi well plate.

9. The method as claimed in claim 1, wherein the aqueous solution is a biological buffer or a culture medium.

10. The method as claimed in claim 9, wherein the culture medium is a viscous liquid or gel.

11. The method as claimed in claim 10, wherein the culture medium comprises proteins, glycoproteins, polysaccharides, methylcellulose, agar, collagen, or other similar gelling agents.

12. The method according to claim 1, wherein the coating material is applied to the interior wall surfaces of the culture vessel using a method selected from:

Application or insertion of pre-formed materials (with or without adhesive);

Application of the material using a physical applicator followed by removal of excess material;

Application by immersion of the vessel into the coating material or a solution thereof, followed by drying;

Application of a melted material followed by cooling and solidification;

Dissolution of the coating material in a suitable solvent and application of this solution, followed by removal of the solvent through evaporation, aspiration, and/or washing;

Application of a material that cures upon exposure to air; and

Application of an agent following addition of the material that causes the material to cure.

13. A method to label cells in a first solution by generating droplets of a second solution containing at least one cell-labelling agent and allowing the droplets of the second solution to contact the surface of the first solution.

14. The method according to claim 13, wherein the cell-labelling agent is a visible light dye, a conjugated antibody that binds to a specific cell-surface antigen or a compound that reacts with a cellular component and can be detected by visible light or fluorescence.

15. The method as claimed in claim 14, wherein the dye is neutral red, Bismarck brown or Nile blue.

16. The method according to claim 14, wherein the antibody is conjugated to a fluorescent entity selected from the group consisting of fluorescein, tetramethylrhodamine isothiocyanate, phycoerythrin, peridinin-chlorophyll-protein complex (PerCP), Cy3, Cy5, C5.5, Cy7, Texas Red, Alexa 488, Alexa Fluor 350, Alexa Fluor 405, Alexa Fluor 430, Alexa Fluor 488, Alexa Fluor 500, Alexa Fluor 514, Alexa Fluor 532, Alexa Fluor 546, Alexa Fluor 555, Alexa Fluor 568, Alexa Fluor 594, Alexa Fluor 610, Alexa Fluor 633, Alexa Fluor 635, Alexa Fluor 647, Alexa Fluor 660, Alexa Fluor 680, Alexa Fluor 700, Alexa Fluor 750 and quantum dots.

17. The method according to claim 14, wherein the compound that reacts with a cellular component is a tetrazolium compounds such as (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (Yellow MTT) and LavaCell™

.

18. The method according to claim 13, wherein the first solution is a viscous liquid or a gel.

19. The method according to claim 18, wherein the first solution is cell culture medium containing methylcellulose, agar or collagen.

20. The method according to claim 13, wherein the droplets are less than 50 μ

L in volume.

21. A vessel for holding liquid comprising a coating material on the interior wall of the vessel, wherein the material provides a receding contact angle between the vessel wall and the liquid of about 90 degrees.

22. The vessel according to claim 21, wherein the hydrophobic material is silicone, EPDM, buna nitrile or petroleum jelly.

23. The vessel according to claim 21, wherein the coating material is a fluoropolymer, or is silicone-based.

24. The vessel according to claim 21, wherein the vessel is a culture dish or well in a multi well plate.

25. The vessel according to claim 21, wherein the liquid is cell culture medium.

26. The vessel according to claim 21, wherein the receding contact angle is between about 75 degrees and about 110 degrees.

27. The vessel according to claim 21, wherein the receding contact angle is between about 80 degrees and about 110 degrees.

28. The vessel according to claim 21, wherein the receding contact angle is between about 85 degrees and about 105 degrees.

29. A use of the vessel according to claim 21 for culturing cells or for performing imaging-based assays.

30. The use according to claim 29, wherein the imaging based assay is selected from colony forming cell (CFC) assays, gene sequencing, combinatorial chemistry, drug discovery and proteomics.

31. A method of imaging cells comprising culturing the cells in the vessel according to claim 21 and imaging the cells.

32. The method according to claim 31, wherein the cells are imaged manually or automatically.

33. The method according to claim 31, wherein the cells are imaged using visible light, ultraviolet light, infrared light and/or fluorescence.

34. The method according to claim 33 wherein the cells are imaged using visible light.

35. The method according to claim 34, wherein the cells are imaged using darkfield mode, brightfield mode, phase contrast or differential interference contrast.

36. The method according to claim 31, wherein the cells are in a cell colony.