Metering doses of sample liquids

• US 7,592,185 B2
• Filed: 08/05/2004
• Issued: 09/22/2009
• Est. Priority Date: 02/17/2004
• Status: Active Grant

• Current Assignee: Molecular BioProducts Incorporated (Perspecta, Inc.)
• Original Assignee: Molecular BioProducts Incorporated (Perspecta, Inc.)
• Inventors: Karg, Jeffrey A., Kroncke, Douglas W.
• Primary Examiner(s): Gordon; Brian R.
• Application Number:
  US10/911,845
• Publication Number:
  US 20050181519A1
• Time in Patent Office: 1,874 Days
• Field of Search: 422/100, 738/633.2, 738/64, 738/640.1, 436/180
• US Class Current: 436/180
• CPC Class Codes: B01F 23/451 by injecting one liquid into another
• CPC Class Codes: B01F 23/483 using water for diluting a liquid ingredient, obtaining a predetermined concentration or making an aqueous solution of a concentrate
• CPC Class Codes: B01F 35/882 using measuring chambers, e.g. volumetric pumps, for feeding the substances
• CPC Class Codes: B01L 2300/06 Auxiliary integrated devices, integrated components
• CPC Class Codes: B01L 2300/0858 Side walls
• CPC Class Codes: B01L 3/021 Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids
• CPC Class Codes: B01L 3/0275 Interchangeable or disposable dispensing tips
• CPC Class Codes: B01L 3/0293 for liquids
• 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 1/38 Diluting, dispersing or mixing samples
• CPC Class Codes: G01N 2035/1032 Dilution or aliquotting
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

A method of metering and mixing a dose of a sample liquid with a diluent liquid includes introducing a sample liquid into a channel defined in a housing. The housing defines a pocket open to the channel and sized to both collect a metered dose of the sample liquid, and to retain the collected dose by capillary force when the channel is emptied. The sample liquid is then removed from the channel under conditions that enable retention of the collected, metered dose of the sample liquid in the pocket. Following a cleaning step, a volume of diluent liquid is introduced into the channel to induce diffusion and mixing of the diluent liquid with the dose of sample liquid to form a mixture.

Claims

1. A method of metering and mixing a dose of a sample liquid with a diluent liquid, the method comprising;

providing a pipette including;

an elongated body having an outer surface and an interior wall, a portion of the interior wall defining an internal cavity open at a lower end of the body;

the body defining an opening in an upper portion of the body, through which air can be drawn from the cavity to draw liquids into the cavity through the lower end of the body;

wherein the portion of the interior wall defining the internal cavity also defines a pocket open to the internal cavity, the pocket radially extending into the portion of interior wall defining the internal cavity, the pocket sized and configured to collect and retain a metered dose of a liquid drawn into the cavity as the cavity is otherwise evacuated;

drawing a sample liquid into the pipette to a level above the pocket such that the pocket collects a metered dose of the sample liquid;

expelling the sample liquid from the pipette under conditions that enable retaining the collected, metered dose of the sample in the pocket by capillary force; and

then

drawing a volume of a diluent liquid into the pipette a sufficient distance to contact the retained dose of sample liquid, to induce diffusion and mixing of the diluent liquid with the dose of sample liquid.

2. The method according to claim 1 further comprising, after expelling the sample liquid from the pipette and before drawing the volume of diluent liquid into the pipette,

drawing a cleaning liquid into the pipette below the pocket;

and then

expelling the cleaning liquid from the pipette to flush any residual sample liquid from surfaces of the interior wall below the pocket.

3. The method according to claim 1 wherein the pipette is formed from a synthetic resin.

4. The method according to claim 1 further comprising dispensing a metered volume of the mixture into a destination well.

5. The method according to claim 4 wherein dispensing the metered volume comprises pneumatically propelling the mixture from the pipette.

6. The method according to claim 4 wherein the pipette further defines a capillary hole between an outer surface of the pipette and the internal cavity.

7. The method according to claim 6 wherein the mixture is propelled from the pipette by forcing a pressurized gas into the internal cavity through the capillary hole.

8. The method according to claim 1 wherein the pocket is sized to collect a metered dose of sample liquid of between about 1 nanoliter and about 10 microliters in volume.

9. The method according to claim 8 wherein the pocket is sized to collect a metered dose of sample liquid of between about 5 nanoliters and about 10 microliters in volume.

10. The method according to claim 1 wherein the internal cavity is narrower at a lower open end of the pipette than in an upper section of the pipette.

11. The method according to claim 10 wherein the pocket is disposed in a portion of the internal cavity wider than the internal cavity at the open end of the pipette.

12. The method according to claim 1 wherein the pocket comprises an upwardly extending notch defined in an interior wall of the pipette.

13. The method according to claim 1 wherein a surface of the pipette defining a lower extent of the pocket is substantially perpendicular to a longitudinal axis of the pipette.

14. The method according to claim 1 wherein the pocket is elongated and parallel to a longitudinal axis of the pipette.

15. The method according to claim 14 wherein the pocket has a length of between about 0.01 and about 1 inch (0.25 and 25 millimeters), as measured parallel to the longitudinal axis of the pipette.

16. The method according to claim 14 wherein the pocket has a width of between about 0.001 and about 0.04 inch (0.025 and 1.02 millimeters).

17. The method according to claim 1 wherein the pocket has a depth of between about 0.001 and about 0.100 inch (0.025 and 2.54 millimeters), measured along a radial line extending from the longitudinal axis of the pipette.

18. The method of claim 1 wherein the volume of diluent liquid introduced into the internal cavity is in the range of between about 1 nanoliter and about 500 microliters.

19. The method according to claim 1 wherein the pipette defines a plurality of said pockets spaced apart from one another.

20. The method according to claim 1 wherein the pocket is formed from a method selected from the group consisting of molding, machining, welding, casting, swaging, cold forming and coating.

21. The method according to claim 1 wherein the pocket is defined in a protrusion extending into the internal cavity.

22. The method according to claim 1 wherein the diluent liquid is moved reciprocally across the pocket multiple times, to induce mixing with the dose of sample liquid.

23. A method of metering and mixing a plurality of doses of a sample liquid with a diluent liquid, the method comprising;

providing an array of pipettes, each pipette comprising

an elongated body having an outer surface and an interior wall, a portion of the interior wall defining an internal cavity open at a lower end of the body;

the body defining an opening in an upper portion of the body, through which air can be drawn from the internal cavity to draw liquids into the cavity through the lower end of the body;

wherein the portion of the interior wall defining the internal cavity also defines a pocket open to the internal cavity, the pocket radially extending into the portion of interior wall defining the internal cavity, the pocket sized and configured to collect a metered dose of a sample liquid;

drawing the sample liquid into the pipettes to a level above the pockets such that each pocket collects a metered dose of the sample liquid;

expelling the sample liquid from the pipettes under conditions that enable retaining the collected, metered doses of the sample in the pockets by capillary force; and

then

drawing a diluent liquid into the pipettes a sufficient distance to contact the retained doses of sample liquid, to induce diffusion and mixing of the diluent liquid with the doses of sample liquid.

24. The method according to claim 23 further comprising, after expelling the sample liquid from the pipettes and before drawing the diluent liquid into the pipettes,

drawing a cleaning liquid into the pipettes below the pockets;

and then

expelling the cleaning liquid from the pipettes to flush any residual sample liquid from surfaces of the interior wall below the pocket.

25. The method according to claim 23 wherein the pipettes are formed from a synthetic resin.

26. The method according to claim 23 further comprising dispensing from each pipette a metered volume of the mixture into a destination well.

27. The method according to claim 26 wherein dispensing the metered volume comprises pneumatically propelling the mixture from the pipettes.

28. The method according to claim 27 wherein each pipette further defines a capillary hole between an outer surface of the pipette and the internal cavity.

29. The method according to claim 28 wherein the mixture is propelled from the pipette by forcing a pressurized gas into the internal cavity through the capillary hole.

30. The method according to claim 23 wherein the pocket is sized to collect a metered dose of sample liquid of between about 1 nanoliter and about 10 microliters in volume.

31. The method according to claim 30 wherein the pocket is sized to collect a metered dose of sample liquid of between about 5 nanoliters and about 10 microliters in volume.

32. The method according to claim 23 wherein the internal cavity is narrower at a lower open end of the pipette than in an upper section of the pipette.

33. The method according to claim 32 wherein the pocket is disposed in a portion of the internal cavity wider than the internal cavity at the open end of the pipette.

34. The method according to claim 23 wherein the pocket comprises an upwardly extending notch defined in an interior wall of the pipette.

35. The method according to claim 23 wherein a surface of the pipette defining a lower extent of the pocket is substantially perpendicular to a longitudinal axis of the pipette.

36. The method according to claim 23 wherein the pocket is elongated and parallel to a longitudinal axis of the pipette.

37. The method of claim 23 wherein the volume of diluent liquid introduced into the internal cavity is in the range of between about 1 nanoliter and about 500 microliters.

38. The method according to claim 23 wherein the pipette defines a plurality of said pockets spaced apart from one another.

39. The method according to claim 23 further comprising dispensing a metered volume of the mixture into an array of liquid-receiving units.

40. The method of claim 39 wherein the array of pipettes is aligned directly above the array of liquid-receiving units.

41. The method according to claim 39 wherein the array of liquid-receiving units is a multi-well container.

42. The method according to claim 41 wherein the multi-well container is selected from the group consisting of a 96-well microtiter plate, a 384-well microtiter plate, and 1536-well microtiter plate.

43. The method according to claim 23 further comprising dispensing a metered volume of the mixture onto a slide.

44. The method according to claim 23 further comprising dispensing a metered volume of the mixture onto an electronic assay reading device.