Gravity-Assisted Mixing Methods

• US 20090136913A1
• Filed: 12/12/2008
• Published: 05/28/2009
• Est. Priority Date: 06/21/2007
• Status: Active Grant

• Current Assignee: Gen-Probe Incorporated (Hologic Incorporated), Qualigen, Inc.
• Original Assignee: Gen-Probe Incorporated (Hologic Incorporated), Qualigen, Inc.
• Inventors: Nelson, Norman C., Scott, Matthew J., Taylor, Jason A., Fan, Sara H., Lee, Richard S., Breidenthal, Scott S.
• Granted Patent: US 8,048,375 B2
• Time in Patent Office: Days
• US Class Current: 435/3
• CPC Class Codes: B01F 2101/23 Mixing of laboratory samples e.g. in preparation of analysing or testing properties of materials
• CPC Class Codes: B01F 2101/33 Mixing compost ingredients or organic waste
• CPC Class Codes: B01F 23/59 Mixing systems, i.e. flow charts or diagrams
• CPC Class Codes: B01F 31/00 Mixers with shaking, oscillating, or vibrating mechanisms
• CPC Class Codes: B01F 31/31 using receptacles with deformable parts, e.g. membranes, to which a motion is imparted
• CPC Class Codes: B01F 31/65 the materials to be mixed being directly submitted to a pulsating movement, e.g. by means of an oscillating piston or air column
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01F 35/2131 Colour or luminescence
• CPC Class Codes: B01L 2200/0621 Control of the sequence of chambers filled or emptied
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2200/14 Process control and prevention of errors
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/123 Flexible; Elastomeric
• CPC Class Codes: B01L 2300/1805 Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• CPC Class Codes: B01L 2300/1822 : using Peltier elements
• CPC Class Codes: B01L 2300/1883 : using thermal insulation
• CPC Class Codes: B01L 2400/0457 : passive flow or gravitation
• CPC Class Codes: B01L 2400/0478 : pistons
• CPC Class Codes: B01L 2400/0481 : squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0655 : pinch valves
• CPC Class Codes: B01L 2400/0677 : phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
• CPC Class Codes: B01L 3/502 : with fluid transport, e.g. in multi-compartment structures centrifugal-type cuvettes G01N21/07; analysis by separation into components G01N30/00; automatic analysers G01N35/00
• 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 3/502761 : 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 3/505 : flexible containers not provided for above
• CPC Class Codes: B01L 7/52 : with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B01L 7/525 : with physical movement of samples between temperature zones
• CPC Class Codes: C12N 15/1006 : by means of a solid support carrier, e.g. particles, polymers
• CPC Class Codes: C12Q 1/6806 : Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6844 : Nucleic acid amplification reactions
• CPC Class Codes: G01N 1/44 : Sample treatment involving radiation, e.g. heat
• CPC Class Codes: G01N 2021/6419 : Excitation at two or more wavelengths
• CPC Class Codes: G01N 2021/6421 : Measuring at two or more wavelengths
• CPC Class Codes: G01N 2021/6432 : Quenching
• CPC Class Codes: G01N 2021/6439 : with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 2021/6463 : Optics
• CPC Class Codes: G01N 2035/00544 : using fluid flow
• CPC Class Codes: G01N 21/01 : Arrangements or apparatus for facilitating the optical investigation
• CPC Class Codes: G01N 21/6428 : Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/645 : Specially adapted constructive features of fluorimeters
• CPC Class Codes: G01N 2201/0627 : Use of several LED's for spectral resolution
• 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: Y10T 137/0391 : Affecting flow by the addition of material or energy
• CPC Class Codes: Y10T 436/143333 : Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/255 : including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

A receptacle having a plurality of interconnected chambers arranged to permit multiple process steps or processes to be performed independently or simultaneously. The receptacles are manufactured to separate liquid from dried reagents and to maintain the stability of the dried reagents. An immiscible liquid, such as an oil, is included to control loading of process materials, facilitate mixing and reconstitution of dried reagents, limit evaporation, control heating of reaction materials, concentrate solid support materials to prevent clogging of fluid connections, provide minimum volumes for fluid transfers, and to prevent process materials from sticking to chamber surfaces. The receptacles can be adapted for use in systems having a processing instrument that includes an actuator system for selectively moving fluid substances between chambers and a detector. The actuator system can be arranged to concentrate an analyte present in a sample. The detector can be used to detect an optical signal emitted by the contents of the receptacle.

Claims

1. A system for mixing substances comprising:

a receptacle having a plurality of interconnected chambers, including a first chamber directly connected to an at least partially compressible second chamber; and

an analyzer supporting the receptacle in an operative position and comprising;

one or more actuators constructed and arranged to apply a compressive force to the second chamber, the receptacle being positioned in the analyzer so that gravity will draw a fluid from the first chamber into the second chamber when fluid communication is established between the first and second chambers; and

a controller programmed to control operation of the one or more actuators and to cause the one or more actuators to;

(a) apply a compressive force to the second chamber, thereby displacing at least a portion of the contents of the second chamber into the first chamber;

(b) remove the compressive force from the second chamber, thereby allowing the displaced contents to flow by gravity from the first chamber to the second chamber without applying a compressive force to the first chamber; and

(c) repeat steps (a) and (b) one or more times.

2. The system of claim 1, wherein the one or more actuators have compression pads that are arranged to generally conform to the shape of the second chamber.

3. The system of claim 1, wherein the one or more actuators are pneumatic actuators.

4. The system of claim 1, wherein, prior to applying the compressive force to the second chamber, the first chamber contains a dried substance, the second chamber contains a liquid substance for reconstituting, dissolving or rehydrating the dried substance, and a connection between the first and second chambers is initially blocked to prevent the liquid substance from coming into contact with the dried substance.

5. The system of claim 4, wherein the dried substance comprises a lyophilized reagent.

6. The system of claim 5, wherein the second chamber further contains an immiscible liquid, the immiscible liquid being non-reactive with the liquid substance.

7. The system of claim 6, wherein the immiscible liquid is an oil.

8. The system of claim 7, wherein the oil is a mineral oil.

9. The system of claim 1, wherein:

each of the chambers of the receptacle is an at least partially compressible chamber to facilitate substance movement between chambers,

the analyzer comprises a plurality of actuators, each of the actuators being associated with at least one of the chambers; and

the controller is programmed to control operation of the actuators and to cause the actuators to move substances among the interconnected chambers by selective application of external force by the actuators to their associated chambers to compress the associated chambers and displace the contents of the chambers into interconnected chambers.

10. The system of claim 1, wherein the receptacle comprises opposed members, at least one of the opposed members comprising a flexible sheet.

11. The system of claim 10, wherein each of the opposed members comprises a flexible sheet.

12. The system of claim 1, wherein an openable connection directly connecting the first and second chambers is altered from a closed state to an open state when the compressive force is applied to the second chamber, the open state of the connection providing fluid communication between the first and second chambers.

13. The system of claim 12, wherein the first and second chambers are adjacent to each other, and wherein the openable connection is an opening blocked by a burstable seal that is altered from the closed state to the open state when the compressive force is applied to the second chamber.

14. The system of claim 13, wherein the burstable seal is a heat seal.

15. The system of claim 12, wherein each of the plurality of interconnected chambers is positioned adjacent to at least one other chamber of the receptacle.

16. The system of claim 12, wherein the analyzer further comprises an actuator adjacent the connection between the first and second chambers, and wherein the controller is programmed to control operation of the actuator to cause the actuator to block the connection between the first and second chambers after step (c).

17. The system of claim 1, wherein the analyzer does not include an actuator associated with the first chamber.

18. The system of claim 17 further comprising:

an actuator system that includes an actuator plate for operatively positioning a plurality of actuators relative to the receptacle, the actuator plate having an opening; and

a detector positioned adjacent the opening to permit the detector to detect a characteristic of the contents of the first chamber.

19. The system of claim 18, wherein the detector is an optical detector.

20. The system of claim 19,wherein the detector is a fluorometer.

21. A method of mixing substances within a receptacle having a plurality of interconnected chambers, the method comprising the steps of:

(a) orienting the receptacle in an analyzer so that gravity can draw the contents of a first chamber of the receptacle into a second chamber of the receptacle when fluid communication is established between the first and second chambers;

(b) establishing fluid communication between the first and second chambers;

(c) applying a substance-moving force to the contents of the second chamber so that the contents of the second chamber are moved into the first chamber;

(d) removing the substance-moving force of step (c) so that gravity draws the contents of the first chamber into the second chamber; and

(e) repeating steps (c) and (d) one or more times.

22. The method of claim 21, wherein the receptacle remains stationary during the method.

23. The method of claim 21, wherein the fluid communication is established by the formation of a narrow opening between the first and second chambers.

24. The method of claim 21, wherein, prior to step (b), the first chamber contains a dried substance and the second chamber contains a liquid substance for reconstituting, dissolving or rehydrating the dried substance.

25. The method of claim 24, wherein the dried substance is a lyophilized substance.

26. The method of claim 24 further comprising the step of reconstituting, dissolving or rehydrating the dried substance contained in the first chamber.

27. The method of claim 24, wherein the second chamber further contains an immiscible liquid, the immiscible liquid being non-reactive with the liquid substance.

28. The method of claim 27, wherein the immiscible liquid is an oil.

29. The method of claim 28, wherein the oil is a mineral oil.

30. The method of claim 21, wherein the substance-moving force is one or more compressive forces applied to a flexible portion of the second chamber.

31. The method of claim 30, wherein the receptacle comprises opposed members configured to define the plurality of interconnected chambers.

32. The method of claim 31, wherein at least one of the opposed members comprises a flexible sheet, the flexible sheet including the flexible portion of the second chamber.

33. The method of claim 32, wherein each of the opposed members comprises a flexible sheet.

34. The method of claim 30, wherein step (b) comprises altering an openable connection between the first and second chambers from a closed state to an open state when the substance-moving force is applied to the second chamber.

35. The method of claim 34, wherein the plurality of interconnected chambers are defined by seals formed between the opposed members.

36. The method of claim 35, wherein the first and second chambers are adjacent to each other, and wherein the openable connection is an opening blocked by a burstable seal that is altered from the closed state to the open state when the substance-moving force is applied to the second chamber.

37. The method of claim 36, wherein the burstable seal is a heat seal.

38. The method of claim 34, wherein the substance-moving force is applied by one or more actuators having compression pads that are arranged to generally conform to the shape of the second chamber.

39. The method of claim 38, wherein the actuators are pneumatic actuators.

40. The method of claim 34, further comprising, after step (e), the step of blocking the connection between the first and second chambers to prevent substance movement between the first and second chambers.

41. The method of claim 40, wherein the connection between the first and second chambers is blocked by one or more compressive forces applied to the connection.

42. The method of claim 21, wherein each of the plurality of interconnected chambers is adjacently positioned next to at least one other chamber of the receptacle.

43. The method of claim 21 further comprising the step of detecting with a detector at least one characteristic of the contents of the first chamber.

44. The method of claim 43, wherein the detector is in a stationary position adjacent the first chamber during the method.

45. The method of claim 44, wherein the detector is a fluorometer.