Methods of manufacturing high-throughput screening systems

• US 6,413,782 B1
• Filed: 03/19/1998
• Issued: 07/02/2002
• Est. Priority Date: 06/28/1996
• Status: Expired due to Term

• Current Assignee: Caliper Holdings Incorporated
• Original Assignee: Caliper Holdings Incorporated
• Inventors: Parce, J. Wallace, Kopf-Sill, Anne R., Bousse, Luc J.
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US09/044,587
• Time in Patent Office: 1,566 Days
• Field of Search: 436/28, 436/29, 436/62, 436/149, 436/163, 436/806, 436/150, 436/514, 435/4, 435/29, 435/40, 435/291, 435/817, 435/810, 204/299.R, 204/180.1, 204/601, 204/451, 422/68, 422/72, 422/73, 422/81, 422/312, 422/82, 422/57, 422/58, 422/102, 422/99, 422/100
• US Class Current: 436/514
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/00828 Silicon wafers or plates
• CPC Class Codes: B01J 2219/00831 Glass
• CPC Class Codes: B01J 2219/00833 Plastic
• CPC Class Codes: B01J 2219/00853 Employing electrode arrangements
• CPC Class Codes: B01J 2219/00952 Sensing operations
• CPC Class Codes: B01J 2219/00961 Temperature
• CPC Class Codes: B01J 2219/0097 Optical sensors
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0418 : electro-osmotic flow [EOF]
• CPC Class Codes: B01L 2400/0487 : fluid pressure, pneumatics
• CPC Class Codes: B01L 3/5025 : for parallel transport of multiple samples
• CPC Class Codes: B01L 3/502715 : characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/50273 : characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• 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: B01L 3/502784 : specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: G01N 27/44791 : Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 33/5008 : for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/5011 : for testing antineoplastic activity
• CPC Class Codes: G01N 33/502 : for testing non-proliferative effects
• CPC Class Codes: G01N 33/5064 : Endothelial cells
• CPC Class Codes: G01N 33/5091 : for testing the pathological state of an organism
• CPC Class Codes: G01N 33/5097 : involving plant cells immunoassays of plant cells G01N33/56961
• CPC Class Codes: G01N 33/5304 : Reaction vessels, e.g. agglutination plates for solid-phase systems G01N33/543
• CPC Class Codes: G01N 33/57415 : of breast
• CPC Class Codes: G01N 35/08 : using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
• CPC Class Codes: Y10S 366/02 : Micromixers: segmented laminar flow with boundary mixing orthogonal to the direction of fluid propagation with or without geometry influences from the pathway
• CPC Class Codes: Y10S 435/81 : Packaged device or kit
• CPC Class Codes: Y10S 435/817 : Enzyme or microbe electrode
• CPC Class Codes: Y10S 436/804 : Radioisotope, e.g. radioimmunoassay
• CPC Class Codes: Y10S 436/805 : Optical property
• CPC Class Codes: Y10S 436/806 : Electrical property or magnetic property
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/110833 : Utilizing a moving indicator strip or tape
• CPC Class Codes: Y10T 436/113332 : with conveyance of sample along a test line in a container or rack
• CPC Class Codes: Y10T 436/117497 : with a continuously flowing sample or carrier stream
• CPC Class Codes: Y10T 436/25 : including sample preparation

Abstract

Microfluidic devices for analyzing a plurality of compounds are manufactured for performing high throughput screening assays. The methods are used to manufacture devices that screen large numbers of different compounds for their effects on a variety of chemical and biochemical systems.

Claims

1. A method of manufacturing an apparatus for screening test compounds for an effect on a biochemical system, comprising:

fabricating a substrate having at least one surface;

fabricating at least two intersecting channels in said surface of said substrate, at least one of said at least two intersecting channels having at least one cross-sectional dimension in the range from about 0.1 to about 500 μ

m;

providing a source of a plurality of different test compounds fluidly connected, via a capillary or microchannel, to a first of said at least two intersecting channels;

providing a source of at least one component of said biochemical system fluidly connected to a second of said at least two intersecting channels;

providing a fluid direction system for flowing said at least one component within said second of said at least two intersecting channels and for introducing said different test compounds from said first to said second of said at least two intersecting channels;

providing a cover mated with said surface; and

optionally fabricating a detection zone in said second channel for detecting an effect of said test compound on said biochemical system.

2. The method of claim 1, wherein said fluid direction system is fabricated to generate a continuous flow of said at least first component along said second of said at least two intersecting channels, and periodically injects a test compound from said first channel into said second channel.

3. The method of claim 1, further comprising providing a source of a second component of said biochemical system, and fabricating a third channel into said surface, said third channel fluidly connecting at least one of said at least two intersecting channels with said source of said second component of said biochemical system.

4. The method of claim 3, wherein said fluid direction system generates a continuous flow of a mixture of said first component and said second component along said second of said at least two intersecting channels, and periodically injects a test compound from said first channel into said second channel.

5. The method of claim 1, wherein said the method comprises:

providing at least three electrodes, each electrode being in electrical contact with said at least two intersecting channels on a different side of an intersection formed by said at least two intersecting channels; and

providing a control system for concomitantly applying a variable voltage at each of said electrodes, whereby movement of said test compounds or said at least first component in said at least two intersecting channels are controlled.

6. The method of claim 1, wherein said substrate is fabricated by etching a glass slide.

7. The method of claim 1, comprising fabricating an insulating layer disposed over said substrate.

8. The method of claim 1, wherein said substrate is fabricated by molding a polymer.

9. The method of claim 1, comprising fabricating a plurality of electrodes in a plurality of reservoirs fluidly connected to one or more of said intersecting channels and providing a control system for concomitantly applying a voltage to each of said electrodes, whereby movement of said first component in said at least two intersecting channels is controlled.

10. The method of claim 1, the method comprising fabricating one or more component for reducing electroosmotic flow, selected from the group consisting of:

a frit on one or more of the electrodes, which frit reduces electroosmotic flow towards the one or more electrodes;

a first channel between at least two of said reservoirs, which first channel limits diffusion of said chemical species, the channel having low electroosmotic flow;

a second channel between at least two of said reservoirs, which second channel limits diffusion of said chemical species, the narrow channel treated to reduce electroosmotic flow;

a filled channel between at least two of said reservoirs, which filled channel comprises a matrix to limit transport of said chemical species through the filled channel, the filled channel thereby having low electroosmotic flow;

a high reservoir having a fluid level higher than at least one low reservoir, which high reservoir is fluidly connected to said low reservoir, which low reservoir comprises an electrode, wherein fluid pressure between the high reservoir and the low reservoir reduces electroosmotic flow towards the electrode; and

,

a dual reservoir system with a first reservoir fluidly connected through a connecting channel to a narrow diameter second reservoir adapted to receive one of the plurality of electrodes, said narrow diameter second reservoir adapted to draw fluid by capillary electrophoresis towards the one electrode, thereby countering electroosmotic flow in the connecting channel.

11. A method of making an apparatus for detecting an effect of a test compound on a biochemical system, comprising:

fabricating a substrate having at least one surface;

fabricating a plurality of reaction channels in said surface;

fabricating at least two transverse channels in said surface, each of said plurality of reaction channels being fluidly connected to a first of said at least two transverse channels at a first point in said reaction channels, and fluidly connected to a second of said at least two transverse channels at a second point in said reaction channels, said at least two transverse channels and said plurality of reaction channels each having at least one cross-sectional dimension in the range from about 0.1 to about 500 μ

m;

providing a source of at least one component of said biochemical system, said source of at least one component of said biochemical system being fluidly connected to each of said plurality of reaction channels;

providing a source of test compounds fluidly connected, via a capillary or microchannel, to said first of said at least two transverse channels;

providing a fluid direction system for controlling movement of said test compound and said at least one component within said at least two transverse channels and said plurality of reaction channels;

providing a cover mated with said surface; and

optionally providing a detection system for detecting an effect of said test compound on said biochemical system.

12. The method of claim 11, wherein the method comprises:

providing a plurality of individual electrodes, each in electrical contact with each terminus of said at least two transverse channels; and

providing a control system for concomitantly applying a variable voltage at each of said electrodes, whereby movement of said test compounds or said at least first component in said at least two transverse channels and said plurality of reaction channels are controlled.

13. The method of claim 11, wherein each of said plurality of reaction channels is fabricated to comprise a bead resting well at said first point in said plurality of reaction channels.

14. The method of claim 11, wherein said source of at least one component of a biochemical system is fabricated in fluid connection to said plurality of reaction channels by fabricating a third transverse channel having at least one cross sectional dimension in a range of from 0.1 to 500 μ

m and being fluidly connected to each of said plurality of reaction channels at a third point in said reaction channels.

15. The method of claim 11, wherein said third point in said reaction channels is intermediate to said first and second points in said reaction channels.

16. The method of claim 15, further comprising fabricating a particle retention zone in each of said plurality of reaction channels, between said third and said second points in said plurality of reaction channels.

17. The method of claim 16, wherein said particle retention zone comprises a particle retention matrix.

18. The method of claim 16, wherein said particle retention zone is fabricated to comprise a microstructural filter element.

19. The method of claim 11, wherein said plurality of reaction channels are fabricated to comprise a plurality of parallel reaction channels fabricated into said surface of said substrate and said at least two transverse channels are connected at opposite ends of each of said parallel reaction channels.

20. The method of claim 11, wherein said at least two transverse channels are fabricated on said surface of said substrate in inner and outer concentric channels, respectively, and said plurality of reaction channels extend radially from said inner concentric channel to said outer concentric channel.

21. The method of claim 20, wherein said detection system comprises a detection window fabricated in said second channel.

22. The method of claim 20, wherein said detection system comprises a fluorescent detection system.

23. The method of claim 11, wherein said substrate is fabricated by etching glass.

24. The method of claim 11, further comprising fabricating an insulating layer over an etched glass substrate.

25. The apparatus of claim 11, wherein said substrate is fabricated by molding a polymer.

26. A method of manufacturing a microfluidic device, the method comprising:

fabricating a body, and at least first, second and third channels disposed in the body, wherein the first, second and third channels communicate at a first intersection, the first channel connecting a source of at least one target component with the first intersection, the second channel connecting a source of a plurality of test compounds with the intersection, and the third channel connecting the first intersection with a detection zone for detecting an effect of the test compound on the target component.