Apparatus for mixing liquids used in countercurrent multiphase liquid separation

• US 5,578,201 A
• Filed: 05/26/1995
• Issued: 11/26/1996
• Est. Priority Date: 07/30/1993
• Status: Expired due to Fees

• Current Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.)
• Original Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.)
• Inventors: King, Robert A., Kopatsis, Alexander D., Deane, Brian A., Feathers, William E., Mrozinski, Peter M., Ladisch, Thomas P., Pih, Norman P., Collier, Charles F.
• Primary Examiner(s): Drodge, Joseph W.
• Application Number:
  US08/451,353
• Time in Patent Office: 550 Days
• Field of Search: 210/511, 210/634, 210/143, 210/141, 210/142, 422/100-104, 422/116, 422/255, 422/269, 422/258, 422/259, 435/177, 435/178, 435/180, 366/131, 366/132, 366/273, 366/241
• US Class Current: 210/142
• CPC Class Codes: B01D 11/04 of solutions which are liquid
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: G01N 1/34 Purifying; Cleaning processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: G01N 2001/4061 Solvent extraction

Abstract

A reusable apparatus for multiphase, multistage countercurrent liquid separation is presented. The apparatus is particular useful in progressive multistage separation of target microorganisms from background organisms and materials with precise transfer, air exclusion, homogeneous mixing, isolation from the ambient atmosphere, in the absence of valves and at ow cost. The structure has at least one chamber in which a piston-like sliding bottom is sealingly moved or braked and includes internal structure for mixing, preferably a magnetic-stainless steel ball which can be reciprocated from the outside to mix the liquid contents. Motion of the bottom is limited by stops. A number of ways to actuate the apparatus are taught.

Claims

1. Apparatus for performing multi-phase liquid separation comprising:

a separation chamber which has an open end and a closed end, said closed chamber end having an inlet passage and an outlet passage;

a sliding bottom inserted into the open end of said separation chamber for sliding motion therein while providing a liquid seal thereto;

means disposed within said separation chamber for mixing liquid within said separation chamber;

removable inlet closing means;

removable outlet closing means; and

means for moving in a selected stroke said sliding bottom toward said closed end and means for selectively preventing said sliding bottom from being moved toward said open end along said stroke at any selected location between said open end and said closed end.

2. The apparatus of claim 1 wherein said separation chamber is charged with a selected volume of two-phase aqueous medium completely filling said chamber between said closed end and said sliding bottom.

3. The apparatus of claim 1 wherein said mixing means is a magnetically-responsive ball disposed in said chamber between said closed end and said sliding bottom.

4. The apparatus of claim 1 wherein said means to move and prevent moving of said sliding bottom is a rod of selected length and diameter to freely enter said chamber through said open end to contact said sliding bottom and selectively move said sliding bottom toward said closed end or selectively hold said sliding bottom against sliding toward said open end by manual or mechanized translation or braking of said rod.

5. The apparatus of claim 1 wherein said means for selectively moving and preventing said sliding bottom from being moved comprises:

sealing means to seal said open end of said chamber;

a source of fluid power;

a valve for said chamber in fluid communication with said source and said chamber through said sealing means; and

means to selectively open and close said valves in selected sequence.

6. The apparatus of claim 1 wherein the internal portion of said chamber wherein said sliding bottom slides is a cylinder.

7. The apparatus of claim 6 wherein said chamber has stop means limiting the motion of said sliding bottom toward said closed end.

8. The apparatus of claim 7 wherein the internal portion of said chamber between said closed end and the internal portion wherein said sliding bottom slides is oval, the intersection of the two portions thereby providing said stop means.

9. The apparatus of claim 1 further comprising:

a storage chamber which has an open end and a closed end, said closed end having an inlet passage and an outlet passage, said storage chamber and said closed and open ends thereof being adjacent to said separation chamber and said open and closed ends thereof, the outlet passage of said separation chamber interconnected by a transfer passage to the inlet passage of said storage chamber;

a sliding bottom inserted into the open end of said storage chamber for sliding motion therein while providing a liquid seal thereto;

means for selectively moving said sliding bottom in said storage chamber toward said closed end and selectively preventing said sliding bottom from being moved toward said open end.

10. The apparatus of claim 9 wherein said separation chamber is charged with a selected volume of two-phase aqueous medium.

11. The apparatus of claim 9 wherein said mixing means is a magnetically-responsive ball disposed in said separation chamber between said closed end and said sliding bottom.

12. The apparatus of claim 9 wherein said means to move and prevent moving of said sliding bottoms are rods of selected length and diameter to freely enter said chambers through said open ends to contact said sliding bottoms and selectively move said sliding bottoms towards said closed ends or selectively hold said sliding bottoms against sliding toward said open ends by manual or mechanized translation or braking of said rods.

13. The apparatus of claim 12 comprising mechanized translation and braking effected by a threaded translating rod driven by an internal nut rotated by a reversible electric stepping motor.

14. The apparatus of claim 9 wherein said means for selectively moving and preventing said sliding bottom from being moved comprises:

means to seal said open ends of each said chamber;

a source of fluid power;

a valve for each said chamber in fluid communication with said source and said chamber through said sealing means; and

means to selectively to open and close said valves in selected sequence.

15. The apparatus of claim 9 wherein said transfer passage is a region extending between said outlet passage of said separation chamber and said inlet passage of said storage chamber and said region is covered with a thin film sealed peripherally around said outlet passage thereof, said region and said inlet passage thereof.

16. The apparatus of claim 15 wherein said thin film is pressed by a pad fixed to a cover removably fastened to said closed ends to close each of said passages during shipping and handling.

17. The apparatus of claim 9 wherein said transfer passage is a groove extending between said outlet passage of said separation chamber and said inlet passage of said storage chamber and said groove is covered with a thin film sealed peripherally around said outlet passage thereof, said groove, and said inlet passage thereof.

18. The apparatus of claim 17 wherein said thin film is pressed by a pad fixed to a cover removably fastened to said closed ends to close each of said passages during shipping and handling.

19. The apparatus of claim 18 wherein said means for selectively moving and preventing moving each of said sliding bottoms are rods of selected length and diameter to freely enter said chambers through said open ends to contact said sliding bottoms and selectively move said sliding bottoms towards said closed ends or selectively hold said sliding bottoms against sliding toward said open ends by manual or mechanized translation or braking means.

20. The apparatus of claim 19 wherein the mixing means comprise a magnetically-responsive ball disposed in each of said separation chambers between said closed ends and said sliding bottoms.

21. The apparatus of claim 20 wherein said separation chambers are charged with selected volumes of two-phase aqueous medium completely filling said chambers between said closed ends and said sliding bottoms and a volume of a liquid sample containing organic matter to be separated is inoculated into said first separation chamber displacing the sliding bottom therein to accommodate said added volume in combination with mixing apparatus comprising:

a cavity for holding said apparatus of claim 20;

a permanent magnet; and

means to reciprocate said magnet in proximity to said magnetically responsive balls.

22. The apparatus of claim 20 wherein said separation chambers are charged with selected volumes of two-phase aqueous medium completely filling said chambers between said closed ends and said sliding bottoms and a volume of a liquid sample containing organic matter to be separated is inoculated into said first separation chamber displacing the sliding bottom therein to accommodate said added volume in combination with actuating apparatus comprising:

a holding cavity; and

means to selectively translate or brake said rods;

whereby an aliquot portion of said sample-containing medium is selectively transferred sequentially from chamber to chamber without reverse flow or unselected overflow to a subsequent chamber; and

said actuating apparatus is selectively disposed relative to gravity such that said aliquot portion is either a top or bottom phase.

23. The apparatus of claim 20 wherein said separation chambers are charged with selected volumes of two-phase aqueous medium completely filling said chambers between said closed ends and said sliding bottoms and a volume of a liquid sample containing organic material to be separated is inoculated into said first separation chamber displacing the sliding bottom therein to accommodate said added volume in combination with mixing and actuating apparatus comprising:

a holding cavity;

means to selectively translate or brake said rods whereby an aliquot portion of said sample-containing medium is transferred sequentially from chamber to chamber without reverse flow or unselected overflow to a subsequent chamber;

a permanent magnet;

means to reciprocate said magnet in proximity to said magnetically-responsive balls; and

wherein said actuating apparatus is selectively disposed relative to gravity such that said aliquot portion is either a top or bottom phase.

24. The apparatus of claim 19 comprising mechanized translation and braking effected by a threaded translating rod driven by an internal nut rotated by a reversible electric stepping motor.

25. The apparatus of claim 9 wherein each said sliding bottom is a cylinder.

26. The apparatus of claim 25 wherein said separation chamber has stop means limiting the motion of said sliding bottom toward said closed end thereof.

27. The apparatus of claim 26 wherein the internal portion of said separation chamber between the closed end and the portion wherein said sliding bottom slides is oval, the intersection of the two portions thereby providing said stop means.

28. The apparatus of claim 9 for performing two-phase aqueous separation wherein:

there is a plurality of separation chambers disposed in sequential, side-by-side relationship and the outlet passage of a first of said plurality of chambers interconnected by a transfer passage to a further one of said separation chambers and so on in sequentia, the outlet passage of the last of said plurality of separation chambers being interconnected by a transfer passage to the inlet passage of said storage chamber.

29. The apparatus of claim 28 wherein said separation chambers are charged with selected volumes of two-phase aqueous medium completely filling said chambers between said closed ends and said sliding bottoms.

30. The apparatus of claim 28 wherein the mixing means comprise a magnetically-responsive ball disposed in each of said separation chambers between said closed ends and said sliding bottoms.

31. The apparatus of claim 28 wherein the internal portion of said chambers wherein said sliding bottoms slide is a cylinder.

32. The apparatus of claim 31 wherein said separation chambers have stop means limiting the motion of said sliding bottoms toward said closed ends.

33. The apparatus of claim 32 wherein the internal portion of said separation chambers between said closed ends and the portion wherein said sliding bottoms slide are oval, the intersection of the two portions thereby providing said stop means.

34. The apparatus of claim 28 wherein said transfer passages are a region extending between each of said outlet and inlet passages of said sequential chambers and said inlet passage of said storage chamber and said region is covered with a thin film sealed peripherally around said outlet passages thereof said region, and said inlet passages thereof.

35. The apparatus of claim 34 wherein said thin film is pressed by a pad fixed to a cover removably fastened to each of said closed ends to close said respective passages during shipping and handling.

36. The apparatus of claim 28 wherein said transfer passages are grooves extending between each of said outlet and inlet passages of said sequential chambers and said grooves are covered with a thin film sealed peripherally around each said individual outlet passages thereof, said grooves, and said inlet passages thereof.

37. The apparatus of claim 36 wherein said thin film is pressed by a pad fixed to a cover removably fastened to each of said closed ends to close said respective passages during shipping and handling.

38. The apparatus of claim 28 wherein said means for selectively moving and preventing said sliding bottoms from being moved comprise:

means to seal said open ends of each said chamber;

a source of fluid power;

a valve for each said chamber in fluid communication with said source and each said chamber through said sealing means; and

means to selectively open and close said valves singly or in combination in selected sequence.

39. The apparatus of claim 38 wherein the mixing means is a magnetically-responsive ball disposed in each of said separation chambers between said closed ends and said sliding bottoms.

40. The apparatus of claim 39 wherein said separation chambers are charged with selected volumes of two-phase aqueous medium completely filling said chambers between said closed ends and said sliding bottoms and a volume of a liquid sample containing organic matter to be separated is inoculated into said first separation chamber displacing the sliding bottom therein to accommodate said added volume in combination with mixing apparatus comprising:

a holding cavity;

a permanent magnet; and

means to reciprocate said magnet in proximity to said magnetically-responsive balls.

41. The apparatus of claim 39 wherein said separation chambers are charged with selected volumes of two-phase aqueous medium completely filling said chambers between said closed ends and said sliding bottoms and a volume of a liquid sample containing organic matter to be separated is inoculated into said first separation chamber displacing the sliding bottom therein to accommodate said added volume in combination with actuating apparatus comprising:

a holding cavity wherein said means to seal said open ends is incorporated in said cavity and whereby said means to selectively to open and close said valves singly or in combination in selected sequence is actuatable to transfer an aliquot portion of said sample-containing medium sequentially from chamber to chamber without reverse flow or unselected overflow to a subsequent chamber; and

said actuating apparatus is selectively disposed relative to gravity such that said aliquot portion is either a top or bottom phase.

42. The apparatus of claim 39 wherein said separation chambers are charged with selected volumes of two-phase aqueous medium completely filling said chambers between said closed ends and said sliding bottoms and a volume of a liquid sample containing organic material to be separated is inoculated into said first separation chamber displacing the sliding bottom therein to accommodate said added volume in combination with mixing and actuating apparatus comprising:

a permanent magnet;

means to reciprocate said magnet in proximity to said magnetically-responsive balls;

a holding cavity wherein said means to seal said open ends is incorporated and whereby said means to selectively to open and close said valves singly or in combination is selected sequence is actuatable to transfer an aliquot portion of said sample-containing medium sequentially from chamber to chamber without reverse flow or unselected overflow to a subsequent chamber; and

said actuating apparatus is selectively disposed relative to gravity such that said aliquot portion is either a top or bottom phase.

43. The apparatus of claim 28 wherein there are three separation chambers.

44. Apparatus for performing multi-phase liquid separation comprising:

a separation chamber which has an open end and a closed end, said closed chamber end having an inlet passage and an outlet passage;

a sliding bottom inserted into the open end of said separation chamber for sliding motion therein while providing a liquid seal thereto;

means for mixing liquid within said separation chamber;

removable inlet closing means;

removable outlet closing means; and

means for moving in a selected stroke said sliding bottom toward said closed end and means for selectively preventing said sliding bottom from being moved toward said open end along said stroke at any selected location between said open end and said closed end, wherein said means for selectively moving and preventing said sliding bottom from being moved comprises;

sealing means to seal said open end of said chamber;

a source of fluid power;

a valve for said chamber in fluid communication with said source and said chamber through said sealing means; and

means to selectively open and close said valve in selected sequence.

45. Apparatus for performing multi-phase liquid separation comprising:

a separation chamber which has an open end and a closed end, said closed chamber end having an inlet passage and an outlet passage;

a sliding bottom inserted into the open end of said separation chamber for sliding motion therein while providing a liquid seal thereto;

means for mixing liquid within said separation chamber;

removable inlet closing means;

removable outlet closing means; and

means for moving in a selected stroke said sliding bottom toward said closed end and means for selectively preventing said sliding bottom from being moved toward said open end along said stroke at any selected location between said open end and said closed end;

a storage chamber which has an open end and a closed end, said closed end having an inlet passage and an outlet passage, said storage chamber and said closed and open ends thereof being adjacent to said separation chamber and said open and closed ends thereof, the outlet passage of said separation chamber interconnected by a transfer passage to the inlet passage of said storage chamber;

a sliding bottom inserted into the open end of said storage chamber for sliding motion therein while providing a liquid seal thereto;

means for selectively moving said sliding bottom in said storage chamber toward said closed end and selectively preventing said sliding bottom from being moved toward said open end.