Robotic microchannel bioanalytical instrument

• US 6,627,446 B1
• Filed: 07/02/1998
• Issued: 09/30/2003
• Est. Priority Date: 07/02/1998
• Status: Expired due to Fees

• Current Assignee: IntegenX Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Amersham Biosciences SV Corp (Thermo Fisher Scientific Incorporated)
• Inventors: Jovanovich, Stevan B., Roach, David J., Loder, Robert T. Jr., Armstrong, Thomas M., Harris, Dennis W., Johnston, Richard F.
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): WRIGHT, PATRICIA KATHRYN
• Application Number:
  US09/109,676
• Time in Patent Office: 1,916 Days
• Field of Search: 422/63, 422/65, 422/66, 422/81, 422/82.01, 422/82.02, 422/82.05, 422/100, 422/101, 422/104, 436/43, 436/44, 436/149, 436/514, 436/174, 436/180, 204/451, 204/601, 204/604, 435/287.2, 435/288.5
• US Class Current: 436/43
• CPC Class Codes: B01L 13/02 for receptacle or instruments
• 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: G01N 2035/00059 vacuum chucks
• CPC Class Codes: G01N 2035/00158 Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 2035/0437 Cleaning cuvettes or reaction vessels
• CPC Class Codes: G01N 27/44704 Details; Accessories
• CPC Class Codes: G01N 27/44743 Introducing samples
• CPC Class Codes: G01N 27/44782 of a plurality of samples
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 35/0099 comprising robots or similar manipulators robots per se B25J
• CPC Class Codes: G01N 35/1074 arranged in a two-dimensional array
• 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 436/25 including sample preparation
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

A substrate with a plurality of microchannels is movably deployed with other movable objects that will load sample into the microchannels, stimulate molecular migration, read the results of the migration, remove and replace the substrate, and prepare for a new run. The other objects include a gripper for engaging and moving the substrate, an electrode array of fine wires suitable for fitting into the microchannels for electromigration, and a scanning detector for reading migration results. A sequence of automatic operations is established so that one substrate after another may be moved into position, loaded with sample, stimulated for molecular migration, read with a beam, and then removed and replaced with a fresh substrate.

Claims

1. Simultaneous chemical analysis in a first microchip substrate, by the method comprising:

a) providing a microchip substrate defining a plurality of microchannels communicating between opposed cathode and anode ports, said microchip further defining for each microchannel a sample port and a waste port communicating across a segment of said microchannel adjacent said cathode port, and movably mounting the microchannel substrate on a first track at a loading station;

b) providing a plurality of sample wells, each of said sample wells containing a sample aliquot, and movably mounting the sample wells on a second track;

c) mounting a multifunctional microchannel loading device on a gantry spanning the first and second tracks, said loading device having a plurality of disengageable pipette tips;

d) robotically moving said loading device between said plurality of sample wells and said sample ports, and robotically picking up a plurality of said sample aliquots with said plurality of pipette tips, said sample aliquots having detectable target molecules with characteristic migration rates;

e) robotically delivering said plurality of said sample aliquots simultaneously to said plurality of sample ports, said robotically delivering step further comprising the step of inserting said pipette tips through a corresponding plurality of apertures defined by a tip guide positioned in overlying registry with said sample ports;

f) repeating steps d) and e) until a desired number of said sample ports have received samples;

g) robotically moving said first microchip substrate away from said loading station, freeing said loading station for accommodating a replacement microchip substrate;

h) causing sample separation in the microchannels of said first microchip substrate until desired sample separations are achieved; and

i) detecting said sample separations in the microchannels at a measurement location.

2. The method of claim 1 further comprising, following step f), robotically replacing said first microchip substrate with said replacement substrate at said loading station.

3. The method of claim 2 wherein robotically replacing said first microchip substrate with said replacement substrate at said loading station includes replacing said first substrate with a replacement substrate which does not have sample included within the substrate.

4. The method of claim 2 further comprising robotically moving said first microchip from said measurement location after detecting said sample separations, freeing said measurement location for said replacement microchip substrate and repeating steps a)-i) using the replacement microchip substrate.

5. The method of claim 4 wherein steps h) and i) are effected using said first microchip substrate at the same time steps d), e), and f) are effected using said replacement microchip substrate.

6. The method of claim 4 further comprising continuously repeating steps a)-h) using a plurality of replacement microchip substrates, wherein steps h) and i) are effected on one of the plurality of said microchip substrates at the same time steps d), e), and f) are effected using another of said microchip substrates.

7. The method of claim 4, further comprising after step f) robotically replacing said sample in said sample wells.

8. The method of claim 1 wherein causing sample separation in the microchannels of said first microchip substrate is effected by robotically moving the chip to place the chip in contact with a plurality of wires such that the contents within a plurality of the individual microchannels on said substrate are brought into contact with an anode wire and a cathode wire and introducing electric current through said microchannel to effect electrophoretic separation.

9. The method of claim 1 wherein said detection is further defined by optically scanning the plurality of microchannels.

10. The method of claim 9 wherein said optically scanning is further defined by confocal optically scanning a plurality of the microchannels.

11. The method of claim 9 wherein providing microchannels in a microchip substrate includes providing microchannels arranged in parallel or at an angle, with a transverse line defining a scan region.

12. The method of claim 1 wherein the step of providing a microchannel loading device comprises providing a plurality of pipettors at spacing equal to the spacing of the sample wells and the inlet ports, said plurality of pipetteors disengageably retaining said plurality of pipette tips.

13. The method of claim 1 wherein providing a microchannel loading device is further defined by providing two parallel tracks having moveable stages, including a first track having a stage for the microchip substrate, and a second track having a stage for the sample wells and for pipettor tips, with the loading device being moveable between the two tracks.

14. The method of claim 1, further comprising the steps of:

disengaging said plurality of pipette tips; and

engaging a plurality of replacement pipette tips in substitution for said plurality of pipette tips.

15. The method of claim 14, wherein said engaging step further comprises the step of:

frictionally engaging and retaining said replacement pipette tips on said loading device.

16. A method for simultaneously separating and detecting DNA sequences of a sample aliquot comprising the steps of:

providing first microchip substrate defining a plurality of elongate converging microchannels, each said microchannel communicating between opposed cathode and anode ports and each substrate defining a sample port and a waste port communicating across a segment of said microchannel adjacent said cathode port and movably positioning said microchip substrate on a first track at a loading station;

providing a plurality of sample wells, each of said sample wells containing a sample aliquot, and movably positioning said sample wells on a second track;

movably mounting a microchannel loading device on a gantry spanning the first and second tracks, said loading device having a plurality of disengageable pipette tips;

robotically drawing, transporting, and dispensing, from an array of robotically replaceable pipette tips provided on said loading device, a sample aliquot from an array of sample aliquot wells to said sample ports of said microchip substrate;

robotically controlling migration of said sample aliquot from said sample port into said segment of said microchannel;

robotically separating said dispensed portion of said sample aliquots within said segments of the microchannels and causing said separated dispensed portions to migrate towards said anode port of the microchannel; and

optically scanning the separated sample at a scanning region of said microchannels and detecting DNA sequences.

17. The method of claim 16, further comprising the step of:

robotically moving said first microchip substrate from said loading station where said sample aliquots are dispensed into said sample ports to an analysis station in registry with a confocal four filament scanner.

18. The method of claim 16, further comprising the step of:

providing a tip guide for precisely guiding said array of pipette tips into fluid communication with said sample ports of said microchip substrate.