Microfabricated apparatus for cell based assays

• US 8,030,062 B2
• Filed: 09/23/2008
• Issued: 10/04/2011
• Est. Priority Date: 04/27/1998
• Status: Expired due to Fees

• Current Assignee: Gyros Patent AB (Mesa Laboratories Inc.)
• Original Assignee: Gyros Patent AB (Mesa Laboratories Inc.)
• Inventors: Andersson, Per, Thomas, Nicholas
• Primary Examiner(s): Beisner; William H
• Application Number:
  US12/236,120
• Publication Number:
  US 20090239292A1
• Time in Patent Office: 1,106 Days
• Field of Search: 435/288.5, 435/297.2, 435/297.5, 422/415, 422/72, 422/506
• US Class Current: 435/288.5
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0806 Standardised forms, e.g. compact disc [CD] format
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/10 Means to control humidity and/or other gases
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0688 surface tension valves, capillary stop, capillary break
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: C12M 23/16 Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: C12M 23/38 Caps; Covers; Plugs; Pouring means
• CPC Class Codes: C12M 25/16 Particles; Beads; Granular material; Encapsulation chemical or physical processes conducted in the presence of fluids and solid particles B01J8/00

Abstract

Apparatus and methods are provided for performing cell growth and cell based assays in a liquid medium. The apparatus comprises a base plate supporting a plurality of micro-channel elements, each micro-channel element comprising a cell growth chamber, an inlet channel and an outlet channel, a cover plate positioned over the base plate to define the chambers and connecting channels. Means are incorporated in the cell growth chambers, for cell attachment and cell growth. In particular, the invention provides a rotatable disc microfabricated for performing cell growth and cell based assays. The apparatus and method can be used for the growth of cells and the detection and measurement of variety of biochemical processes and products using non-invasive techniques, that is techniques which do not compromise the integrity or viability of cells.

Claims

1. An apparatus microfabricated for performing cell growth in a liquid medium, said apparatus comprising:

a) a base plate supporting a plurality of micro-channel elements, each comprising a cell growth chamber, an inlet channel, and an outlet channel;

b) a cover plate positioned over said base plate said cover plate extending over said elements so as to define said chambers and channels; and

c) a hydrophobic valve in at least one of said chambers or said channels that is defined by positioning said cover plate over said base plate, wherein said valve comprises a localized region of hydrophobicity within said chamber or said channel, wherein the hydrophobic valve prevents liquid flow until subjected to the application of a defined force,

wherein at least some of said micro-channel elements each comprises one or more assay chambers for performing assays involving cellular constituents and connected in line between said cell growth chamber and said outlet channel.

2. The apparatus of claim 1 further comprising means, incorporated in said cell growth chamber, for cell growth, wherein one or more components of the apparatus are constructed of a gas permeable film or membrane such that CO₂-buffered media can be used in the cell growth chamber and gas may permeate to and from the cells for their metabolism during growth.

3. The apparatus of claim 2, wherein said means for cell growth comprises at least a portion of a surface of said cell growth chamber that is treated to allow for cell attachment.

4. The apparatus of claim 3, wherein said surface is negatively charged surface.

5. The apparatus of claim 3, wherein said surface comprises a coating of polylysine, collagen or fibronectin.

6. The apparatus of claim 2, wherein said means for cell growth comprises one or more microcarrier beads located in said cell growth chamber, wherein each of said microcarrier beads provides for cell attachment.

7. The apparatus of claim 1 further comprising a suspension of cells to be grown in each of said cell growth chambers.

8. The apparatus of claim 7, wherein said cells require attachment to said cell growth chambers for growth.

9. The apparatus of claim 1, wherein said base plate comprises a rotatable disc which is microfabricated to provide a sample introduction port located towards the center of the disc and connected to an annular sample reservoir, and wherein said micro-channel elements are radially dispersed on said disc with their respective input channels connected to receive sample from said reservoir.

10. The apparatus of claim 1, wherein said cover plate is fabricated from a gas permeable plastic material.

11. The apparatus of claim 10, wherein the plastic material is a silicone polymer, polyurethane or polytetrafluoroethylene.

12. The apparatus of claim 11, wherein the silicone polymer is polydimethylsiloxane.

13. The apparatus of claim 1, wherein said cell growth chamber comprises raised moulded features disposed on the base portion of said cell growth chamber to form pillars.

14. The apparatus of claim 1, wherein the cross-sectional area of said inlet channel is greater than that of said outlet channel.

15. The apparatus of claim 14, wherein the cross-sectional area of said outlet channel is between 0.99 and 0.01 times that of said inlet channel.

16. The apparatus of claim 1, wherein each assay chamber is connected to each other and to said cell growth chamber by an intermediate channel in the order of:

inlet channel, cell growth chamber, intermediate channel, assay chamber, outlet channel, and wherein the cross-sectional areas of the respective channels reduce progressively from the inlet channel to the outlet channel.

17. The apparatus of claim 16, wherein the cross-sectional area of each intermediate channel and the outlet channel is between 0.99 and 0.01 times that of the immediately preceding upstream channel.

18. The apparatus of claim 16, wherein there is provided in or on an interior surface of one or more of said chambers a layer comprising a scintillant substance.

19. The apparatus of claim 18, wherein the layer comprising a scintillant substance comprises a binding moiety bound thereto, said binding moiety being a member of a specific binding pair selected from the group consisting of biotin, streptavidin, protein A, antibodies, lectins, hormone-receptors, nucleic acid probes, and DNA-binding proteins.