Microfabricated apparatus for cell based assays

• US 6,632,656 B1
• Filed: 11/17/2000
• Issued: 10/14/2003
• Est. Priority Date: 04/27/1998
• Status: Expired due to Term

• Current Assignee: Gyros Patent AB (Mesa Laboratories Inc.)
• Original Assignee: Gyros AB (Mesa Laboratories Inc.)
• Inventors: Andersson, Per, Thomas, Nicholas
• Primary Examiner(s): Beisner, William H.
• Application Number:
  US09/673,169
• Time in Patent Office: 1,061 Days
• Field of Search: 435/30, 435/32, 435/34, 435/33, 435/288.5, 435/297.5, 435/287.3, 435/288.7, 435/395, 435/401, 435/402, 422/72
• 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 for supplying liquid sample thereto and an outlet channel for removal of liquid sample therefrom, a cover plate positioned over the base plate to define the chambers and connecting channels, the cover plate being supplied with holes to provide access to the 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 a 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. A method for determining the effect of a test substance on a cellular activity or physical parameter, which method comprises:

providing an apparatus having

i) a base plate supporting a plurality of micro-channel elements, each comprising a cell growth chamber, an inlet channel for supplying liquid sample thereto and an outlet channel for removal of liquid sample therefrom;

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

said cover plate being supplied with holes to provide access to said channels; and

iii) means, incorporated in said cell growth chambers, 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 oxygen may permeate to the cells for their metabolism during growth;

b) introducing said cells into said apparatus and causing said cells to be transported to one or more cell growth chambers in said apparatus, wherein said cells adhere to one or more cell growth chambers;

c) culturing said cells in one or more cell growth chambers;

d) providing one or more samples of test substances whose effect upon the cells is to be measured under conditions so as to cause said cells to be exposed to said substances, wherein step d) is performed subsequently to step c); and

e) determining the effect of the test substances on said cells by means of optical detection.

2. A method according to claim 1 wherein there are provided following step d) one or more assay reagents and dispersing said reagents to one or more reaction chambers in said apparatus.

3. A method according to claim 2 wherein at least one of said assay reagents is labelled with a detectable label selected from the group consisting of fluorescent labels, chemiluminescent labels, bioluminescent labels, enzyme labels and radioactive labels.

4. The method according to claim 1, wherein the gas permeable film or membrane is a gas permeable plastic material.

5. The method according to claim 4, wherein the gas permeable plastic material is a silicone polymer, polyurethane or polytetrafluroethylene.

6. The method according to claim 5, wherein the silicone polymer is polydimethylsiloxane.

7. A method for measuring a cellular analyte, which method comprises:

providing the apparatus having

i) a base plate supporting a plurality of micro-channel elements, each comprising a cell growth chamber, an inlet channel for supplying liquid sample thereto and an outlet channel for removal of liquid sample therefrom;

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

said cover plate being supplied with holes to provide access to said channels; and

iii) means, incorporated in said cell growth chambers, 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 oxygen may permeate to the cells for their metabolism during growth;

b) introducing said cells into said apparatus and causing said cells to be transported to one or more cell growth chambers in said apparatus, wherein said cells adhere to one or more cell growth chambers;

c) culturing said cells in one or more cell growth chambers;

d) providing one or more assay reagents and dispersing said reagents to one or more chambers in said apparatus, wherein step d) is performed subsequently to step c); and

e) measuring the cellular analyte by optical means.

8. A method according to claim 7 wherein at least one of said assay reagents is labelled with a detectable label selected from the group consisting of fluorescent labels, chemiluminescent labels, bioluminescent labels, enzyme labels and radioactive labels.