Automated apparatus for separating a biological sample from a two dimensional electrophoresis gel

• US 6,652,724 B2
• Filed: 05/18/2001
• Issued: 11/25/2003
• Est. Priority Date: 04/04/2001
• Status: Expired due to Fees

• Current Assignee: Large Scale Proteomics Corporation
• Original Assignee: Large Scale Proteomics Corporation
• Inventors: Michael, Samuel, Goodman, Jack, Anderson, N. Leigh
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): QUAN, ELIZABETH S
• Application Number:
  US09/859,664
• Publication Number:
  US 20020146832A1
• Time in Patent Office: 921 Days
• Field of Search: 204/462, 204/613, 204/466, 204/616, 204/467, 204/618, 204/414, 204/464, 204/463, 738/63, 738/633.3, 738/633.2, 435/287.1, 435/287.3, 422/63, 422/99-104, 436/43, 901/18, 901/15, 901/17
• US Class Current: 204/613
• CPC Class Codes: G01N 2001/288 Filter punches
• CPC Class Codes: G01N 27/44739 Collecting the separated zones, e.g. blotting to a membrane or punching of gel spots
• CPC Class Codes: Y10T 436/11 Automated chemical analysis
• CPC Class Codes: Y10T 436/25 including sample preparation
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

An automated high-throughput system for excising spots or samples from an electrophoresis slab gel includes a computer controlled robotic arm assembly and a sample plate handling assembly for supplying a sample plate to a loading station. The computer is connected to a scanner and imaging device to identify selected sample locations on the slab gel and to direct the robotic arm to the selected locations for excising the gel spots. The cutting assembly includes a removable tray for supporting the slab gel during the cutting process and is coupled to the automated sample plate handling assembly. The sample plate handling assembly delivers a multiwell plate to the cutting assembly for receiving the gel spots. The removable tray cooperates with a scanner for identifying protein spots and includes a positioning device to position the tray in the scanner and the cutting assembly in selected locations to coordinate the scanned image with the cutting process.

Claims

1. An apparatus for excising a plurality of samples from an electrophoresis gel, said apparatus comprising:

a base having a work surface with a loading station dimensioned to support a sample plate having a plurality of sample-receiving wells;

a gel support member removably coupled to said base, said support member comprising a removable tray having a substantially planar surface for supporting said gel in a substantially flat condition and having a coupling member capable of coupling said tray to a robotic tray-manipulating member;

a computer controlled robotic arm assembly having an operating head for excising a plurality of predetermined samples from said gel while said gel is supported on said gel support member and transferring said predetermined samples to a respective well of said sample plate, said robotic arm assembly having a first arm with a first end and a second end, a first motor coupled to said first end of said first arm and oriented to pivot said first arm about an axis perpendicular to a plane of said base, a second motor coupled to said second end of said first arm, a second arm having a first end and second end, said first end of said second arm being coupled to said second motor and being pivotable about an axis perpendicular to said base, and wherein said operating head is coupled to said second end of said second arm;

a microprocessor operatively coupled to said robotic arm for controlling movement of said robotic arm, said microprocessor being programmed to receive a signal for identifying said predetermined samples on said gel, actuating said robotic arm to excise said predetermined samples, and transferring said samples to said respective well of said sample plate; and

an imaging device for obtaining an image of said gel, identifying a plurality of sample locations on said gel, and producing an image signal corresponding to said sample locations, said imaging device being separate from said base and robotic arm of said apparatus and being operatively coupled to said microprocessor whereby said microprocessor receives said image signal and actuates said robotic arm to excise said sample locations.

2. The apparatus of claim 1, wherein said coupling member of said tray has an aperture for coupling with said tray-manipulating assembly.

3. The apparatus of claim 1, wherein said tray has a top surface, and wherein said substantially planar surface is defined by a flat plate coupled to said top surface.

4. The apparatus of claim 3, wherein said plate is a glass plate.

5. The apparatus of claim 4, wherein said glass plate defines a support surface for supporting said gel while excising said samples.

6. The apparatus of claim 1, wherein said tray has a first end for receiving and supporting said gel, a second end opposite said first end, a first side edge and a second side edge, said tray further having a first area spaced between said first end and said second end, said first area being dimensioned to receive a gel handling device.

7. The apparatus of claim 6, wherein said gel is coupled to said gel handling device, said gel handling device being dimensioned to fit in said first area to enable said gel to lay directly on said planar surface of said tray.

8. The apparatus of claim 6, wherein said first area includes at least one aperture.

9. The apparatus of claim 8, wherein said first area of said tray has two spaced apart apertures dimensioned to receive a coupling assembly of a robotic arm.

10. The apparatus of claim 6, wherein said first and second sides of said tray include an inwardly facing notch dimensioned to support said gel handling device.

11. The apparatus of claim 6, wherein said first area is recessed with respect to said planar surface.

12. The apparatus of claim 6, wherein said first side edge of said tray includes an outwardly facing first recess, and wherein said housing includes a first moveable coupling member for engaging said recess to locate said tray in said base in a selected location.

13. The apparatus of claim 12, wherein said first coupling member is a spring biased detent.

14. The apparatus of claim 6, wherein said planar surface is spaced from said first side edge and said second side edge, said tray further comprising a plurality of liquid channels extending between said first side edge and said planar area and between said second side edge and said planar surface.

15. The apparatus of claim 1, wherein said imaging device is an imaging scanner having an imaging area dimensioned to receive said removable gel support member.

16. The apparatus of claim 1, further comprising a storage assembly for storing a plurality of said sample plates, said storage assembly having a conveyor extending between said storage assembly and said top surface of said base.

17. The apparatus of claim 16, wherein said storage assembly includes a supply magazine for storing said sample plates and a receiving magazine for receiving sample plates from said base.

18. The apparatus of claim 17, wherein said storage assembly is operatively coupled to said microprocessor, and wherein said microprocessor is programmed to dispense a sample plate from said supply magazine to said loading station, actuate said robotic arm to transfer said samples into a respective well of said sample plate and to transfer said sample plate from said loading station to said receiving magazine.

19. The apparatus of claim 1, wherein said microprocessor identifies a location of said predetermined samples in said gel based on polar coordinates.

20. The apparatus of claim 19 wherein said recessed area of said tray has a depth substantially equal to a thickness of said first clamping jaw.

21. The apparatus of claim 1, further comprising a gel clamp having first and second pivotally connected clamping jaws each having a gripping end, an electrophoresis gel having one end gripped between said first and second gripping ends, and wherein said tray has a planar area supporting said gel in a flat condition and having a recessed area having a dimension to receive said first clamping jaw whereby said gel lays flat on said planar area.

22. The apparatus of claim 1, wherein said base included two parallel guide rails spaced apart a distance to receive and position said removable try in an operating position and where said base includes a microswitch operatively connected said microprocessor and produces a positioning signal indicating said tray is in said operating condition.

23. An apparatus for excising a plurality of biological samples from an electrophoresis gel, said apparatus comprising:

a gel support member having a planar surface with a dimension for supporting an electrophoresis gel in a substantially flat condition;

a base having a work surface with a loading station dimensioned to removably receive and support said gel support member for said electrophoresis gel and a sample plate having a plurality of sample-receiving wells;

a scanning device separate from said base, said scanning device having a dimension to receive said gel support member and said electrophoresis gel for capturing an image of said gel and producing an image signal corresponding to said image, said gel support member being movable between said scanning device and said base;

an automated sample plate handling assembly coupled to said base, said plate handling assembly including a supply magazine for containing a plurality of empty sample plates, a receiving magazine for receiving filled sample plates, and a conveyor for sequentially conveying a sample plate from said supply magazine to said loading station and for conveying said sample plate from said loading station to said receiving magazine;

a robotic arm having an operating head for excising a plurality of predetermined samples from said gel and for transferring said excised samples to a respective well of a sample plate positioned in said loading station; and

a microprocessor operatively connected to said robotic arm for controlling movement of said robotic arm and being operatively connected to said scanning device, said microprocessor being programmed to receive said image signal from said scanning device and identifying said predetermined samples on said gel and actuating said robotic arm to excise said respective predetermined samples and transferring said predetermined samples to said respective well of said sample plate, said microprocessor further being operatively connected to said sample plate handling assembly for actuating said handling assembly to deliver said empty sample plates sequentially from said supply magazine to said loading station and for conveying said sample plate from said loading station to said receiving magazine after receiving said predetermined samples.

24. The apparatus of claim 23, further comprising an electrophoresis gel, wherein said gel support member is a tray having a first end receiving and supporting said gel, a second end opposite said first end, a first side edge and a second side edge, said tray further having a first recessed area spaced from said first end and dimensioned to receive a gel handling device.

25. The apparatus of claim 24, wherein said gel handling device is a clamp having a first and a second clamping jaw pivotally connected together and where said gel is clamped between said jaws, and wherein said first jaw of said clamp is dimensioned to fit in said first area to enable said gel to lay directly on said tray.

26. The apparatus of claim 25, wherein said first and second sides of said tray include an inwardly facing notch dimensioned to support said gel clamp.

27. The apparatus of claim 24, wherein said first area includes at least one aperture.

28. The apparatus of claim 27, wherein said first area of said tray has two spaced apart apertures dimensioned to receive a coupling assembly of a robotic arm.

29. The apparatus of claim 24, wherein said first side edge of said tray includes an outwardly facing first recess, and wherein said base includes a first moveable coupling member for engaging said recess to locate said tray on said housing in a selected location.

30. The apparatus of claim 29, wherein said coupling member is a spring biased detent.

31. The apparatus of claim 24, wherein said planar area is spaced from said first side edge and said second side edge, said tray further comprising at least one liquid channel extending between said first side edge and said planar area and between said second side edge and said planar area.

32. The apparatus of claim 23, wherein said base includes two parallel guide rails spaced apart a distance to removably receive said gel support member and position said gel support member is an operating position in said base, and where said base includes a microswitch operatively connected to said microprocessor and produces a positioning signal indicating said tray is in said operating condition.

33. The apparatus of claim 23, wherein said microprocessor identifies a location of said predetermined samples in said gel based on polar coordinates.

34. An apparatus for excising a plurality of samples from an electrophoresis gel, said apparatus comprising:

a base having a work surface with a loading station dimensioned to support a sample plate having a plurality of sample-receiving wells;

a gel support tray removably coupled to said base, said tray having a top surface, with a first end and a second end and a support plate coupled to said first end of said top surface for supporting said gel in a substantially flat condition, and a recessed area at said second end and having a dimension for receiving a gel handling device that is removably coupled to said gel, whereby said gel handling device fits in said recessed area to enable said gel to lay flat directly on said support plate, said base having first and second parallel guide rails for guiding said gel support tray to an operating position for excising said samples from said gel,

a computer controlled robotic arm assembly having an operating head for excising a plurality of predetermined samples from said gel while said gel is supported on said gel support member and said tray is in said operating position, and for transferring said predetermined samples to a respective well of said sample plate; and

a microprocessor operatively coupled to said robotic arm for controlling movement of said robotic arm, said microprocessor being programmed to receive a signal for identifying said predetermined samples on said gel, actuating said robotic arm to excise said predetermined samples, and transferring said samples to said respective well of said sample plate.

35. The apparatus of claim 34, wherein said second end of said tray includes at least one aperture for coupling said tray to a robotic tray-manipulating assembly.

36. The apparatus of claim 34, wherein said support plate is a glass plate.

37. The apparatus of claim 34, wherein said recessed area includes at least one aperture.

38. The apparatus of claim 34, wherein said recessed area of said tray has two spaced apart apertures dimensioned to receive a coupling assembly of a robotic arm.

39. The apparatus of claim 34, wherein said first and second sides of said tray include an inwardly facing notch in said recessed area dimensioned to support said gel handling device.

40. The apparatus of claim 34, wherein said tray has a first side edge and a second side edge cooperating with said guide rails, said first side edge having a first recess, and wherein said base includes a first movable coupling member for engaging said recess to locate said tray on said base in said operating position, and where said coupling member is operatively connected to said microprocessor whereby said microprocessor operates said robotic arm only when said tray is in said operating position.

41. The apparatus of claim 40, wherein said first coupling member is a spring biased detent and is movable from a first position to a second position received in said first recess.

42. The apparatus of claim 40, wherein said support plate is spaced from said first side edge and said second side edge, said tray further comprising a plurality of liquid channels extending between said first side edge and said planar area and between said second side edge and said support plate.

43. The apparatus of claim 40, wherein said first movable coupling member includes a microswitch operatively connected to said microprocessor, said microswitch position signal indicating said tray is in said operating position.

44. The apparatus of claim 34, wherein said microprocessor identifies a location of said predetermined samples based on polar coordinates.