Dilution and mixing cartridge

• US 5,104,813 A
• Filed: 04/13/1989
• Issued: 04/14/1992
• Est. Priority Date: 04/13/1989
• Status: Expired due to Term

• Current Assignee: Roche Diagnostics Operations Incorporated (Roche Holding AG)
• Original Assignee: Biotrack, Inc. (Novartis Ag)
• Inventors: Gorin, Michael, Besemer, Don, Gibbons, Ian
• Primary Examiner(s): NOT, DEFINED
• Assistant Examiner(s): Trembley, Theresa A.
• Application Number:
  US07/337,286
• Time in Patent Office: 1,097 Days
• Field of Search: 356/246, 422/50, 422/58, 422/61, 422/100, 422/102, 422/82.05, 422/106, 422/105, 436/165, 436/174, 436/180, 436/179
• US Class Current: 436/179
• 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: B01L 3/502723 characterised by venting arrangements
• CPC Class Codes: G01N 1/38 Diluting, dispersing or mixing samples
• CPC Class Codes: G01N 33/04 Dairy products
• CPC Class Codes: Y10T 436/25625 Dilution
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

A dilution and mixing cartridge is provided that allows single (or multiple) dilutions of a sample with a diluent in a disposable cartridge in which a measurement, such as optical density, is made. Addition of sample to the device automatically measures the sample, and addition of diluent automatically causes a fixed ratio of sample and diluent to enter a receiving chamber, in which mixing and measurement can take place. A number of preferred embodiments are provided in which the diluent application site is located at a vertical height below the sample application site, flow directing chambers are provided to control flow of desired volumes of sample or diluted sample at the appropriate locations in the device and to direct excess sample and diluent to waste locations, or control mechanism are provided at flow directing chambers and other internal locations to verify correct operation of the device.

Claims

1. A dilution and mixing cartridge for automatically measuring and diluting a liquid sample with a liquid diluent, comprising:

in a housing,

sample receiving means for receiving a sample,

a flow directing chamber, comprising a first internal chamber in said housing,

sample flow means for delivering sample from said sample receiving means to said flow directing chamber,

diluent receiving means for receiving a diluent,

diluent flow means for delivering diluent from said diluent receiving means to said flow directing chamber,

a receiving chamber, comprising a first vented internal chamber in said housing having a first volume,

a measuring chamber, comprising a second internal chamber in said housing having a second volume smaller than said first volume, wherein said measuring chamber connects said flow directing chamber to said receiving chamber and said measuring chamber is capable of delivering sample and diluent from said flow directing chamber to said receiving chamber solely by capillary and gravitational forces,

a stop flow junction located at the end of said measuring chamber, wherein said stop flow junction with conforms to the surface tension characteristics of said sample so as to provide sufficient back pressure resulting from contact between said sample and wall means of said housing at said stop flow junction to prevent sample from flowing through said stop flow junction in absence of diluent but allowing flow through said stop flow junction when said diluent receiving means receives diluent and diluent enters said flow directing chamber, and

waste means for emptying said flow directing chamber after sample has entered said flow directing chamber and said sample has filled said measuring chamber, wherein said waste means operates by capillary and gravitational forces,

whereby diluent added to said diluent receiving means flows through said flow directing chamber and expels sample from said measuring chamber into said receiving chamber, thereby causing a fixed ratio of sample to diluent to be delivered to said receiving chamber.

2. The cartridge of claim 1, wherein said flow directing chamber comprises:

(1) an exit for said measuring chamber and said waste means at gravitationally low points in said flow directing chamber, and

(2) entrances for said sample and diluent flow means gravitationally above said exit and said waste means.

3. The cartridge of claim 2, wherein said entrance for said sample flow means is located above said measuring chamber exit and positioned so that flow of sample into said flow directing chamber reaches said measuring chamber exit before reaching said waste means.

4. The cartridge of claim 2, wherein said flow directing chamber comprises a floor sloping downward to said exit and said waste means.

5. The cartridge of claim 2, wherein said waste means comprises an exit in said flow directing means having a cross section smaller than the cross section of said exit leading to said measuring chamber.

6. The cartridge of claim 2, wherein said flow directing chamber further comprises a vent at a gravitationally high location in said chamber.

7. The cartridge of claim 1, wherein said waste means comprises:

(1) a waste chamber, comprising a second vented internal chamber in said housing, and

(2) waste flow means for delivering sample and diluent from said flow directing chamber to said waste chamber by capillary and gravitational forces.

8. The cartridge of claim 1, wherein said waste means comprises a wicking element.

9. The cartridge of claim 1, wherein said sample receiving means is an external chamber on a surface of said housing, said diluent receiving means is (1) an external chamber on a surface of said housing or (2) a third vented internal chamber in said housing, and said diluent receiving means is located at a height below the sample receiving means.

10. The cartridge of claim 9, wherein said diluent receiving means comprises said third vented internal chamber containing a diluent in a sealed container.

11. The cartridge of claim 10, wherein said diluent receiving means chamber has an upper surface, said sample receiving means comprises a chamber open to the external environment and having an upper lip, said upper surface of said diluent receiving means chamber being lower than said upper lip of said sample receiving means.

12. The cartridge of claim 1, wherein said housing further comprises a mixture isolating chamber hydrostatically connected to the receiving chamber, whereby a representative sample of a mixture of sample and diluent in said receiving chamber can be isolated in said mixture isolating chamber.

13. The cartridge of claim 12, further comprising valve means selectively preventing flow from the receiving chamber to the mixture isolating chamber.

14. The cartridge of claim 13, wherein said valve comprises a resilient blocking member capable of obstructing said mixture flow means.

15. The cartridge of claim 14, wherein said blocking member is biased to allow flow through said mixture flow means in the absence of externally applied forces on said blocking member.

16. The cartridge of claim 14, wherein said valve further comprises an access port sealed with a resilient seal through which an external force can be applied to said blocking member to impart motion to said blocking member.

17. The cartridge of claim 12, wherein said mixture isolating chamber comprises:

a valve controlling exit of said mixture form said receiving chamber,

a mixture flow directing chamber, comprising a third internal chamber in said housing,

mixture flow means for delivering mixture from said valve to said mixture flow directing chamber,

second diluent receiving means for receiving a second diluent,

second diluent flow means for delivering said second diluent from said second diluent receiving means to said mixture flow directing chamber,

a mixture receiving chambe, comprising a fourth vented internal chamber in said housing having a third volume,

a mixture measuring chamber, comprising a fourth internal chamber in said housing having a fourth volume smaller than said second volume or said third volume, wherein said mixture measuring chamber connects said mixture flow directing chamber to said mixture receiving chamber and said mixture measuring chamber is capable of delivering mixture and second diluent from said mixture flow directing chamber to said mixture receiving chamber solely by capillary and gravitational forces,

a second stop flow junction located at the intersection of said mixture measuring chamber and said mixture receiving chamber, wherein said second stop flow junction conforms to the surface tension characteristics of said mixture so as to provide sufficient back pressure resulting from contact between said mixture and wall means of said housing at said second stop flow junction to prevent mixture from flowing through said second stop flow junction in absence of second diluent but allowing flow through said stop flow junction when said second diluent receiving means receives second diluent and second diluent enter said mixture flow directing chamber, and

mixture waste means for emptying said second flow directing chamber after mixture has entered said mixture flow directing chamber and said mixture has filled said mixture measuring chamber, wherein said waste means operates by capillary and gravitational forces,

whereby second diluent added to said second diluent receiving means after mixture has filled said mixture measuring chamber flows through said mixture flow directing chamber and expels mixture from said mixture measuring chamber into said mixture receiving chamber, thereby causing a fixed ratio of mixture to second diluent to be delivered to said mixture receiving chamber.

18. A sample dilution and mixing system, comprising:

the cartridge of claim 17, and

a monitor comprising;

means for detecting a reaction in the mixture receiving chamber of said cartridge, and

registration means for holding said cartridge in registration with said detecting means.

19. The system of claim 18, wherein said monitor comprises second system control means adjacent said mixture flow directing chamber when said cartridge is present in said registration means to verify presence of liquid at said second flow directing chamber.

20. The system of claim 19, wherein said second system control means comprises a light source and a detector located on opposite sides of said cartridge when said cartridge is present in said registration means, wherein light emitted by said light source passes through said second flow directing chamber at a light path location and is detected by said detector.

21. The system of claim 20, wherein said light path is located between said mixture measuring chamber exit and said waste means.

22. The system of claim 21, wherein said light path is located to intersect a bottom surface of said second flow directing chamber.

23. The system of claim 18, wherein said monitor comprises third system control means adjacent said mixture measuring chamber when said cartridge is present in said registration means for verifying presence of liquid in said mixture measuring chamber.

24. The system of claim 23, wherein said third system control means comprises a light source and a detector located on opposite sides of said cartridge when said cartridge is present in said registration means, wherein light emitted by said light source passes through said mixture measuring chamber at a light path location and is detected by said detector.

25. The system of claim 24, wherein said light path is located adjacent said second stop flow junction.

26. The system of claim 18, wherein said monitor selects a program by which to operate said valve means by detecting a signal from said cartridge when said cartridge is inserted into said monitor.

27. The system of claim 26, wherein said signal is a bar code.

28. A sample dilution and mixing system, comprising:

the cartridge of claim 1, and

a monitor comprising;

means for detecting a reaction in the receiving chamber of said cartridge, and

registration means for holding said cartridge in registration with said detecting means.

29. The system of claim 28, wherein said detecting means comprises a light source and a light detector arranged to detect light passing through said receiving chamber.

30. The system of claim 29, wherein said flow directing chamber comprises:

exits for said measuring chamber and said waste means at gravitationally low points in said flow directing chamber, and

entrances for said sample and diluent flow means gravitationally above said exit and said waste means.

31. The system of claim 30, wherein said monitor further comprises system control means adjacent said flow directing chamber when said cartridge is present in said registration means to verify presence of liquid at said flow directing chamber.

32. The system of claim 31, wherein said system control means comprises a light source and a detector located on opposite sides of said cartridge when said cartridge is present in said registration means, wherein light emitted by said light source passes through said flow directing chamber at a light path location and is detected by said detector.

33. The system of claim 32, wherein said light path is located between said measuring chamber exit and said waste means.

34. The system of claim 30, wherein said light path is located to intersect a bottom surface of said flow directing chamber.

35. The system of claim 28, wherein said diluent receiving means in said cartridge comprises a diluent chamber containing said diluent enclosed in a rupturable container and said diluent chamber comprises an access port sealed with a flexible sealing member.

36. The system of claim 35, wherein said monitor comprises programmed diluent control means and said container is ruptured under control of said diluent control means.

37. The system of claim 35, wherein said diluent control means comprises a rupturing pin, said pin being capable of motion into said diluent chamber through said access port under control of said programmed diluent control means without rupturing said flexible sealing member, whereby motion of said pin into said diluent chamber ruptures said container and allows said diluent to enter said diluent flow means.

38. In a dilution and mixing cartridge for automatically diluting a liquid sample with a liquid diluent comprising, in a housing, a sample receiving site, a diluent receiving site, a sample measuring chamber, means for delivering sample and diluent to said sample measuring chamber from said sample receiving site and said diluent receiving site, respectively, a stop flow junction terminating said sample measuring chamber and conforming to the surface tension characteristics of the sample so as to provide sufficient back pressure resulting from contact between said sample and wall means of said housing at said stop flow junction to prevent sample from flowing through said stop flow junction in absence of diluent but allowing flow through said stop flow junction when said diluent receiving means receives diluent, a receiving chamber for receiving a mixture of sample and diluent from said measuring chamber, and means for delivering sample and diluent from said sample measuring chamber to said receiving chamber, the improvement which comprises:

a flow directing chamber, comprising an internal chamber in said housing, wherein said means for delivering sample to said sample measuring chamber and said means for delivering diluent to said sample measuring chamber both enter said flow directing chamber and an exit in said flow directing chamber leads to said sample measuring chamber; and

waste means for emptying said flow directing chamber after sample has entered said flow directing chamber and said sample has filled said measuring chamber, said waste means being located in said flow directing means and operating by capillary and gravitational forces.

39. In a dilution and mixing cartridge for automatically diluting a liquid sample with a liquid diluent comprising a sample receiving site, a diluent receiving site, a sample measuring chamber, means for delivering sample and diluent to said sample measuring chamber from said sample receiving site and said diluent receiving site, respectively, a stop flow junction terminating said sample measuring chamber and conforming to the surface tension characteristics of the sample so as to provide sufficient back pressure resulting from contact between said sample and wall means of said housing at said stop flow junction to prevent sample from flowing through said stop flow junction in absence of diluent but allowing flow through said stop flow junction when said diluent receiving means receives diluent, a receiving chamber for receiving a mixture of sample and diluent from said measuring chamber, and means for delivering sample and diluent from said sample measuring chamber to said receiving chamber, the improvement which comprises:

locating said diluent receiving means gravitationally below said sample receiving means and providing means for preventing hydrostatic pressure from developing between said sample receiving means and said stop flow junction in the absence of diluent.

40. The cartridge of claim 39, wherein said means for preventing hydrostatic pressure is a vented, non-capillary chamber located between said sample receiving means and said stop flow junction.

41. A method for diluting a liquid sample with a liquid diluent, comprising:

sequentially supplying an unmeasured volume of said sample and a measured or unmeasured volume of said diluent to a liquid flow directing chamber, wherein (1) a first portion of said sample entering said flow directing chamber is diverted to a measuring chamber having a first volume, flow of sample through said measuring chamber terminating when said sample reaches a stop flow junction, said stop flow junction conforming to the surface tension characteristics of said sample so as to provide sufficient back pressure resulting from contact between said sample and wall means of said housing at said stop flow junction to prevent sample from flowing through said stop flow junction, the remainder of said sample being diverted by said flow directing chamber to a waste chamber, and (2) a first portion of said diluent entering said flow directing chamber is diverted to said measuring chamber, wherein hydrostatic pressure from said diluent expels said sample in said measuring chamber into a receiving chamber along with said first portion of said diluent, whereby a fixed ratio of sample to diluent is delivered to said receiving chamber.

42. The method of claim 41, further comprising the step of opening first valve means selectively controlling passage of said mixture from said receiving chamber to a hydrostatically connected mixture isolating chamber, wherein a hydrostatically determined portion of said first mixture enters said mixture isolating chamber.

43. The method of claim 42, wherein said device further comprises a second receiving chamber in fluid receiving relationship to said mixture isolating chamber.

44. The method of claim 43, wherein a second diluent is added to said device, wherein said second diluent expels a portion of said mixture in said mixture isolating chamber into said second receiving chamber along with a portion of said diluent, whereby a fixed ratio of said mixture and said second diluent are delivered by capillary and gravitational forces to said second receiving chamber.