Microfluidic microorganism detection system

• US 20030023149A1
• Filed: 07/01/2002
• Published: 01/30/2003
• Est. Priority Date: 06/29/2001
• Status: Abandoned Application

• Current Assignee: Carlo D. Montemagno, Hercules Neves
• Original Assignee: Carlo D. Montemagno, Hercules Neves
• Inventors: Montemagno, Carlo D., Neves, Hercules
• Application Number:
  US10/187,309
• Publication Number:
  US 20030023149A1
• Time in Patent Office: Days
• US Class Current: 600/300
• CPC Class Codes: B01L 2200/025 Align devices or objects to ensure defined positions relative to each other
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 3/502746 characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• 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: G01N 15/1245 Devices using more than one aperture
• CPC Class Codes: G01N 15/134 Devices using two or more apertures
• CPC Class Codes: G01N 2015/1254 Electrodes
• CPC Class Codes: G01N 2015/135 Electrodes
• CPC Class Codes: G01N 33/1893 using flow cells

Abstract

A system and method for detecting microorganisms and abiotic or biotic contaminants in fluids, including food and potable and environmental waters. Various embodiments of the system include a capillary transport element and a microsensor element. The capillary transport element isolates and purifies the targeted substance. The microsensor element includes a channel with electrodes for detecting dielectric properties of the targeted substance. Both the transport element and the microsensor may be fabricated using micromachining or nanofabrication techniques. In one embodiment, an output of the transport element is coupled to the input of a microsensor. The targeted substance can be retained in a storage vessel for further analysis. The system may be integrated into a handheld device using disposable cartridges for detecting different microorganisms or contaminants.

Claims

1. A microorganism detection system comprising:

a flow cell having a passageway including an antibody for said microorganism immobilized on an interior surface of said passageway, said passageway adapted to isolate said microorganism; and

a microchip sensor adapted to electronically detect a dielectric property of said isolated microorganism.

2. The system of claim 1 wherein said passageway is adapted to isolate a pathogenic microorganism.

3. The system of claim 1 wherein said passageway is adapted to isolate an oocyst.

4. The system of claim 1 wherein said passageway is adapted to isolate an oocyst of C. parvum bacterium.

5. The system of claim 1 further comprising a vessel adapted for retaining said isolated microorganism.

6. The system of claim 1 wherein said flow cell is adapted for isolating abiotic contaminants.

7. The system of claim 1 wherein said flow cell is adapted for isolating biotic contaminants.

8. The system of claim 1 wherein said microchip sensor is adapted for isolating abiotic contaminants.

9. The system of claim 1 wherein said microchip sensor is adapted for determining viability of said isolated microorganism.

10. The system of claim 1 wherein said microchip sensor is adapted for determining a dielectric constant of said isolated microorganism.

11. The system of claim 1 wherein said flow cell is adapted for concentrating said microorganism by capillary action.

12. The system of claim 1 wherein said binding partner includes an immobilized monoclonal antibody, polyclonal antibody, or binding fragment thereof, that binds to said microorganism.

13. The system of claim 1 wherein said binding partner includes an immobilized APTase, RNA APTase, or binding fragment thereof, that binds to said microorganism.

14. An apparatus for isolating a component of a fluid, said apparatus comprising:

an input port;

a first planar element having an input edge, a first output edge, and a first plurality of fluid pathways on a first surface of said first planar element between said first input edge and said first output edge, said first input edge of said planar element being coupled to said first input port;

a binding partner that selectively binds to said component, said binding partner immobilized on at least a subset of said first plurality of fluid pathways;

an output port coupled to said first output edge of said first planar element; and

a sealing surface sealably coupled to said first surface of said first planar element;

wherein fluid traversing said first planar element is communicated through said first plurality of said fluid pathways by said planar element and said sealing surface.

15. The apparatus of claim 14 wherein said first planar element comprises silicone elastomer.

16. The apparatus of claim 14 wherein said sealing surface comprises glass, an acrylic polymer, silicon or silicone.

17. The apparatus of claim 14 wherein said pathways of said first plurality of fluid pathways have a depth of about 20 to 40 μ

m.

18. The apparatus of claim 14 wherein said first planar element is hermetically bonded to said sealing surface.

19. The apparatus of claim 14 wherein said binding partner comprises an APTase.

20. The apparatus of claim 14 wherein said binding partner comprises a population of antibodies.

21. The apparatus of claim 14 wherein said first planar element performs affinity purification.

22. The apparatus of claim 14 wherein said first plurality of fluid pathways simulate porous media.

23. The apparatus of claim 14 wherein said first plurality of fluid pathways define a serpentine network of pathways.

24. The apparatus of claim 14 further comprising a second planar element having a second input edge, a second output edge, and a second plurality of fluid pathways on a first surface of said second planar element between said second input edge and said second output edge, said first surface of said second planar element coupled to said first surface of said first planar element and further wherein said binding partner is immobilized on at least a subset of said second plurality of fluid pathways and wherein fluid traversing said first planar element and said second planar element is communicated through said first plurality of fluid pathways and said second plurality of fluid pathways.

25. A method of manufacturing an isolator assembly, said method comprising:

receiving a micromachined mold having a plurality of pathways;

applying a silicone elastomer to said mold;

curing said silicone elastomer for a predetermined time;

removing said silicone elastomer from said mold;

immobilizing a binding partner for a predetermined microorganism to a portion of the silicone elastomer corresponding to the plurality of pathways of the mold; and

bonding a sealing surface to said silicone elastomer.

26. The method of claim 25 wherein applying a silicone elastomer to said mold comprises pouring a silicone elastomer over said mold.

27. The method of claim 25 wherein curing said silicone elastomer for a predetermined time comprises curing with heat.

28. The method of claim 25 wherein curing said silicone elastomer comprises curing by heating to a temperature between 55°

C. and 65°

C.

29. A microsensor comprising:

a fluid inlet for receiving a sample fluid including one or more discrete subunits;

a channel coupled to said fluid inlet, said channel including an interior surface coupled to an orifice, wherein said orifice is adapted for sequentially passing a single discrete subunit;

a first electrode coupled to said orifice;

a second electrode coupled to said orifice, said second electrode electrically isolated from said first electrode; and

a fluid outlet coupled to said channel;

wherein an electrical signal applied to said first electrode is capacitatively communicated to said second electrode as a function of said single discrete subunit within said orifice.

30. The microsensor of claim 29 wherein said channel includes a convergent section.

31. The microsensor of claim 29 wherein said channel includes a divergent section.

32. The microsensor of claim 29 wherein said first electrode and said second electrode are substantially planar.

33. The microsensor of claim 29 wherein said first electrode and said second electrode are substantially parallel.

34. The microsensor of claim 29 wherein said first electrode and said second electrode are fabricated of metal.

35. The microsensor of claim 29 wherein said first electrode and said second electrode are fabricated of gold.

36. The microsensor of claim 29 wherein said orifice has a first and second setting, wherein at a first setting, said orifice has a dimension greater than at a second setting.

37. The microsensor of claim 29 wherein said orifice is variable and subject to control by a user.

38. The microsensor of claim 29 wherein said first electrode and said second electrode generate an electric field within said orifice.

39. The microsensor of claim 29 wherein said orifice has a first side and a second side oriented opposite said first side and further wherein said first electrode is positioned on said first side and said second electrode is positioned on said second side.

40. A system for determining a dielectric property for a pathogen, said system comprising:

fluid intake means adapted to receive a sample fluid, said sample fluid including one or more discrete pathogen subunits, said fluid intake means adapted to receive a predetermined quantity of sample fluid;

first orifice means adapted for passing a single said discrete pathogen subunit, said first orifice means coupled to said fluid intake means;

electrode means adapted for generating an electric field in said first orifice means, said electric field transmitted through a sample fluid within said first orifice means;

signal generating means adapted for generating an input electrical signal and applying said input electrical signal to said electrode means; and

processor means adapted for receiving an output electric signal from said electrode means and determining a dielectric property of said subunit in said first orifice means based on said input electric signal and said output electric signal.

41. The system of claim 40 further comprising a fluid reservoir means coupled to said first orifice means and adapted for receiving said subunit from said first orifice means.

42. The system of claim 40 further comprising channel means coupled to said fluid intake means and said first orifice means, wherein said channel means further includes a convergent section.

43. The system of claim 42 wherein said channel means is adapted for isolating said single subunit of said pathogen.

44. The system of claim 40 wherein said signal generating means and said processor means are adapted for determining a dielectric constant, a dielectric loss, a dielectric breakdown voltage, a dielectric strength, or a dielectric absorption of said pathogen.

45. The system of claim 40 further comprising:

display means adapted for providing visual data to a human operator of said system, said display means coupled to said first orifice means, said electrode means, said signal generating means and said processor means;

user operable control means coupled to said first orifice means, said electrode means, said signal generating means, said processor means, said display means and said user operable control means adapted for facilitating interaction by said human operator;

battery means adapted for supplying power to said first orifice means, said electrode means, said signal generating means, said processor means, said display means, and said user operable control means;

housing means adapted for housing said first orifice means, said electrode means, said signal generating means, said processor means, said display means, said user operable control means and said battery means.

46. The system of claim 40 further comprising:

second orifice means adapted for passing a single subunit of said pathogen, said second orifice means coupled to said fluid intake means; and

second electrode means adapted for generating a second electric field in said second orifice means, said second electric field transmitted through a sample fluid within said second orifice means;

and further wherein said signal generating means are adapted for generating a second input electrical signal and applying said second input electrical signal to said second electrode means; and

further wherein said processor means are adapted for receiving a second output electric signal from said second electrode means and determining a dielectric property of said single subunit of said pathogen in said second orifice means based on said second input electric signal and said second output electric signal.

47. The system of claim 40 further comprising a user-replaceable cartridge, wherein said cartridge carries said first orifice means and said electrode means.

48. A method of detecting a targeted substance, comprising:

introducing a sample fluid suspected of including said targeted substance to a capillary system having an interior surface, said interior surface including an immobilized binding partner that binds with a subunit of said target substance;

releasing said subunit from said immobilized binding partner;

exposing said released subunit to an electromagnetic field within an orifice, said orifice adapted for sequentially passing a single subunit; and

determining a dielectric property for said subunit in said orifice.

49. The method of claim 48 further comprising determining viability of said subunit based on said dielectric property.

50. The method of claim 48 further comprising retaining said subunit in a vessel.

51. The method of claim 48 wherein introducing a sample fluid includes introducing liquid water, liquid food, body fluid, or an atmospheric gas.