Automated molecular biology laboratory

• US 5,443,791 A
• Filed: 08/07/1992
• Issued: 08/22/1995
• Est. Priority Date: 04/06/1990
• Status: Expired due to Term

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: The Perkin-Elmer Corporation Applied Biosystems Division
• Inventors: Metzker, Michael L., Ting, Chen-Hanson, Lachenmeier, Eric, Guiremand, Harry A., Brennan-Marquez, Thomas, Cathcart, G. Richard, Hane, Marianne, Kronick, Melvyn N., Mordan, William J., Hoff, Louis B., Shigeura, John, Mayrand, Paul E., Bridgham, John A., Golda, George S., McBride, Lincoln J., Whiteley, Norman M., Keith, Douglas H.
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): Soderquist, Arlen
• Application Number:
  US07/927,254
• Time in Patent Office: 1,110 Days
• Field of Search: 73/864.11, 73/864.24, 73/864.25, 215/247, 364/188, 137/846, 422/63, 422/65, 422/67, 422/99, 422/100, 422/102, 422/104, 422/106, 435/809, 435/296, 436/54, 436/180, 436/526, 901/46
• US Class Current: 422/65
• CPC Class Codes: B01F 2101/23 Mixing of laboratory samples e.g. in preparation of analysing or testing properties of materials
• CPC Class Codes: B01L 3/50825 Closing or opening means, corks, bungs closures for containers B65D; means for removing stoppers B67B7/02
• CPC Class Codes: G01N 2035/00079 Evaporation covers for slides
• CPC Class Codes: G01N 2035/00237 Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 2035/00356 Holding samples at elevated temperature (incubation)
• CPC Class Codes: G01N 2035/00435 Refrigerated reagent storage
• CPC Class Codes: G01N 2035/0403 Sample carriers with closing or sealing means
• CPC Class Codes: G01N 2035/0475 electric, e.g. stepper motor, solenoid
• CPC Class Codes: G01N 2035/0491 Position sensing, encoding; closed-loop control
• CPC Class Codes: G01N 2035/1013 Confirming presence of tip
• CPC Class Codes: G01N 2035/102 Preventing or detecting loss of fluid by dripping
• CPC Class Codes: G01N 2035/1034 Transferring microquantities of liquid
• CPC Class Codes: G01N 2035/1058 for mixing
• CPC Class Codes: G01N 2035/106 by sucking and blowing
• CPC Class Codes: G01N 35/0098 involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C
• CPC Class Codes: G01N 35/0099 comprising robots or similar manipulators robots per se B25J
• CPC Class Codes: G01N 35/028 having reaction cells in the form of microtitration plates
• CPC Class Codes: G01N 35/1004 Cleaning sample transfer devices
• CPC Class Codes: G01N 35/109 with two horizontal degrees of freedom
• CPC Class Codes: Y10S 435/809 Incubators or racks or holders for culture plates or containers
• CPC Class Codes: Y10T 436/119163 : with aspirator of claimed structure
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

A liquid-handling instrument has a worksurface with registration for modular stations to support containers of liquid, pipette apparatus with a pipette tip coupled to a sensing circuit, a robotic translation system for moving the pipette tip, and a control system with an iconic user interface for programming and editing. A gauge block registered on the worksurface provides for calibration using the sensing tip, and register cavities on the worksurface provide for modular stations. There is a wash station fop the pipette tip on the worksurface. An automated laboratory based on the liquid-handling system has heating and cooling and a sealable incubation station as well as a magnetic separation station. Methods are disclosed using the apparatus to convey droplets of liquid, to aspirate with minimum tip contamination, to mix liquids in containers, and to validate the worksurface. A duck-billed closure is disclosed for minimizing evaporation and cross-contamination during processing, and is a part of a container disclosed for storing and transporting liquids.

Claims

1. A liquid-handling instrument for transferring liquid from one container to another comprising:

a stationary worksurface having machined registration elements for accurately locating and supporting a modular container station for holding said containers of liquid;

a pipette subsystem for aspirating and dispensing liquid, said pipette subsystem comprising a pipette tip coupled to a capacitance sensor;

a robotic translation subsystem for moving said pipette tip to all containers on said worksurface and into and out of said containers;

a gauge block securely fastened to and accurately positioned on said worksurface relative to said registration elements, and having surfaces substantially orthogonal to directions of movement of said pipette tip by said robotic translation subsystem; and

a control subsystem for automatically managing said liquid transfers;

wherein said control subsystem controls said robotic translation subsystem to move said pipette tip to each of said surfaces on said gauge block wherein the pipette tip and the capacitance sensor sense the position of the machined surface of said gauge block thereby establishing a home position on said worksurface for said liquid transfers.

2. An instrument as in claim 1 wherein said registration elements comprise a cavity for positioning the modular container station, said cavity positioned accurately relative to said gauge block, such that a first modular station may be removed and a second modular station substituted therefor while maintaining known dimensions from containers in said stations to the known position of said gauge block.

3. An instrument as in claim 1 wherein said pipette subsystem further comprises a first syringe pump operable by said control subsystem at a first rate for aspirating and dispensing liquids with a first degree of accuracy and a second syringe pump operable by said control subsystem at a second rate for aspirating and delivering liquids with a second degree of accuracy, said first and second syringe pumps being commonly connected to said pipette tip.

4. An instrument as in claim 1 wherein said robotic translation subsystem comprises a robot having a carriage over said stationary worksurface for carrying said pipette tip, said carriage being translatable over the area of said worksurface and also translatable toward and away from said worksurface.

5. An instrument as in claim 4 wherein said robot is a cartesian robot having three directions of travel, two in a horizontal plane and the third direction vertical.

6. An instrument as in claim 5 wherein said robot has three principal drives, and said drives are powered by electric motors.

7. An instrument as in claim 1 wherein said control subsystem comprises an iconic interface having user-selectable icons for programming a protocol and for entering values for control variables, said icons representing specific steps in said protocol and arrangeable in series to determine action sequences, said icons being expandable-in-place to show other steps that an icon comprises.

8. An instrument as in claim 1 further comprising a tip washing station located on said stationary worksurface, and said tip washing station comprises;

a fountain for enclosing said pipette tip during washing; and

a well surrounding said fountain such that liquid flowing from said fountain flows into said well, said well having a drain disposed to communicate unwanted material to an external location.

9. An instrument as in claim 1 wherein at least one of said containers supported by said worksurface comprises closure means having a flexible duck-billed portion extending toward the interior of said container.

10. An instrument as in claim 9 wherein said container comprises an upper rim and said closure means comprises an annular cavity portion surrounding said duck-billed portion for sealing to said upper rim.

11. An automated laboratory for transferring liquids from one container to another and performing a chemistry protocol comprising:

a stationary worksurface having machined registration elements are accurately locating and supporting a modular container station for holding said containers of liquid;

a pipette subsystem for aspirating and dispensing liquid, said pipette subsystem comprising a pipette tip coupled to a capacitance sensor;

a robotic translation subsystem for moving said pipette tip to all containers on said worksurface and into and out of said containers;

a heating subsystem for heating liquids in said containers;

a cooling subsystem for cooling liquids in said containers;

a gauge block securely fastened to and accurately positioned on said worksurface and having surfaces substantially orthogonal to directions of movement of said pipette tip by said robotic translation subsystem; and

a control subsystem for automatically managing said chemistry protocol;

wherein said control subsystem controls said robotic translation subsystem to move said pipette tip to each of said surfaces on said gauge block wherein the pipette tip and the capacitance sensor sense the position of the machined surface of said gauge block thereby establishing a home position on said worksurface for said liquid transfers.

12. An automated laboratory as in claim 11 further comprising an incubation station heatable by said heating subsystem and coolable by said cooling subsystem, said incubation station having cavities coated with a chemically inert coating, and a latching lid with a sealing surface such that with said lid closed and latched, said cavities are individually sealed.

13. An automated laboratory as in claim 12 wherein said cavities each have a machined upper lip such that force exerted on said lid by latching said lid is concentrated on said machined upper lips of said cavities to facilitate sealing.

14. An automated laboratory as in claim 12 wherein said cavities are machined into a metal block, and said metal block is securely fastened to and accurately positioned on said worksurface and removable, such that a block of cavities may be removed and replaced with a new block of cavities.

15. An automated laboratory as in claim 12 wherein said cavities have a lower cylindrical portion for holding a sample and an upper conical portion to provide additional volume to hold liquid aspirated during a cleaning procedure.

16. An automated laboratory as in claim 11 further comprising a magnetic system for passing a magnetic field through containers supported by said worksurface to separate paramagnetic particles from a liquid in said containers, said magnetic system comprising a station having two rows of containers for holding liquid, and a magnetic bar supported on an elevator such that said magnetic bar may be selectively elevated and withdrawn from between said rows of containers.

17. An automated laboratory as in claim 16 wherein said magnetic bar comprises rare-earth magnetic material.

18. An automated laboratory as in claim 11 wherein said registration elements comprise a cavity for positioning the modular container station, said cavity positioned accurately relative to said gauge block, such that a first modular station may be removed and a second modular station substituted therefor while maintaining known dimensions from containers in said stations to the known position of said gauge block.

19. An automated laboratory as in claim 11 wherein said pipette subsystem further comprises a first syringe pump operable by said control subsystem at a first rate for aspirating and dispensing liquids with a first degree of accuracy and a second syringe pump operable by said control subsystem at a second rate for aspirating and delivering liquids with a second degree of accuracy, said first and second syringe pumps being commonly connected to said pipette tip.

20. An automated laboratory as in claim 11 wherein said robotic translation subsystem comprises a robot having a carriage over said stationary worksurface for carrying said pipette tip, said carriage being translatable over the area of said worksurface and also translatable toward and away from said worksurface.

21. An automated laboratory as in claim 20 wherein said robot is a cartesian robot having three directions of travel, two in a horizontal plane and the third direction vertical.

22. An automated laboratory as in claim 21 wherein said robot has three principal drives, one for each of said three directions of travel, and said drives are powered by electric motors.

23. An automated laboratory as in claim 11 wherein said control subsystem comprises an iconic interface having user-selectable icons for programming a protocol and for entering values For control variables, said icons representing specific steps in said protocol and arrangeable in series to determine action sequences, said icons being expandable-in-place to show other steps that comprise art icon.

24. An automated laboratory as in claim 11 further comprising a tip-washing station located on said stationary worksurface, and said tip-washing station comprises;

a fountain for enclosing said pipette tip during washing; and

a well surrounding said fountain such that liquid flowing from said fountain flows into said well, said well having a drain disposed to communicate unwanted material to an external location.

25. An automated laboratory as in claim 11 wherein at least one of said containers supported by said worksurface comprises closure means having a flexible duck-billed portion extending toward the interior of said container.

26. An automated laboratory as in claim 25 wherein said container comprises an upper rim and said closure means comprises an annular cavity portion surrounding said duck-billed portion for sealing to said upper rim.

27. An automated laboratory as in claim 11 wherein said control subsystem comprises control routines with steps for performing automated specific gene detection.

28. An automated laboratory in claim 11 wherein said control subsystem comprises control routines with steps for performing automated nucleic acid sequence detection.

29. An automated laboratory in claim 11 wherein said control subsystem comprises control routines with steps for performing automated fluorescent labelling of nucleic acids.

30. A method for transferring liquid by a robotic translation system from a first container holding a first volume of liquid to a second container holding a second volume of liquid comprising steps of:

aspirating a third volume of liquid from said first volume of liquid in said first container with a pipette having a pipette tip and a capacitance sensor for sensing the position of said pipette tip relative to proximate surfaces;

moving said pipette tip away from said first container by action of said robotic translation system;

dispensing a first droplet of liquid from said pipette tip such that said first droplet depends from said pipette tip but does not separate therefrom, said first droplet having a known droplet volume;

moving said first droplet with said pipette tip by said robotic translation system until said first droplet touches the surface of said second volume of liquid, stopping translation when said capacitance sensor signals contact, and allowing said first droplet to become confluent with said second volume of liquid;

withdrawing said pipette tip from said second volume of liquid;

dispensing a second droplet of liquid from said pipette tip such that said second droplet of liquid depends from said pipette tip but does not separate therefrom, said second droplet having a volume equal to the volume of said first droplet;

moving said second droplet with said pipette tip by said robotic translation system until said second droplet touches the surface of said second volume of liquid, stopping translation when said capacitance sensor signals contact, and allowing said second droplet to become confluent with said second volume of liquid; and

repeating the steps of dispensing droplets and transferring the droplets to said second volume of liquid until a predetermined volume of liquid is dispensed and transferred to said second volume of liquid.

31. A method for checking for positions of elements on a worksurface in a liquid-handling instrument having a pipette subsystem, said pipette subsystem comprising a pipette tip coupled to a capacitance sensor for sensing position of said pipette tip relative to proximate surfaces, said pipette tip movable over said worksurface by a robotic translation subsystem, said method comprising the steps of:

moving said pipette tip in a pattern and at a height over said worksurface such that said pipette tip will not contact any surface if every part is in its proper place;

stopping translation of said pipette tip if said pipette tip contacts any surface as signalled by said capacitance sensor; and

activating a signal that a part is out of position.

32. A method for mixing liquids in a liquid handling instrument having a pipette subsystem with a pipette tip movable over a worksurface by a robotic translation subsystem, said method comprising the steps of:

moving said pipette tip by said robotic translation subsystem to immerse said pipette tip in a volume of liquid in a container;

aspirating liquid into said pipette ending with said pipette tip immersed in said volume of liquid in said container;

dispensing said aspirated liquid into said container while moving said pipette tip in a pattern encompassing substantially the space occupied by said volume of liquid in said con ruiner; and

repeating the steps of aspirating and dispensing while moving said tip in said pattern to thoroughly mix the contents in said container.