Apparatus and methods for using biological material to discriminate an agent

• US 20060073539A1
• Filed: 08/06/2002
• Published: 04/06/2006
• Est. Priority Date: 08/06/2001
• Status: Abandoned Application

• Current Assignee: Vanderbilt University
• Original Assignee: Vanderbilt University
• Inventors: Wikswo, John P., Prokop, Ales, Cliffel, David, Baudenbacher, Franz J., Balcarcel, R. Robert, Monroe, Todd, Eklund, Sven, McGuinness, Owen, Werdich, Andreas Augustinus, Stremler, Mark Andrew, Bapty, Theodore A., Yuansheng, Yang
• Application Number:
  US10/212,918
• Publication Number:
  US 20060073539A1
• Time in Patent Office: Days
• US Class Current: 435/29
• CPC Class Codes: C12M 41/32 of substances in solution
• CPC Class Codes: G01N 2015/1006 for cytology
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: G01N 33/5023 on expression patterns
• CPC Class Codes: G01N 33/5038 involving detection of metabolites per se
• CPC Class Codes: G01N 33/5041 involving analysis of members of signalling pathways
• CPC Class Codes: Y10T 436/115831 Condition or time responsive

Abstract

An apparatus and methods for using biological material to discriminate an agent. In one embodiment of the present invention, the method includes the steps of providing at least one cell, exposing at least one cell to an agent, measuring the response of the cell to the agent in terms of a physical quantity related to at least one of the cellular physiological activities of the cell, and identifying the agent from the measured response. The method further includes the step of quantifying the agent from the measured response.

Claims

1. A method for using biological material to discriminate an agent, comprising the steps of:

a. constructing a matrix of biological signatures having a plurality of elements, wherein each element represents a biological signature of one of a plurality of cells in response to an agent, each biological signature being one of a plurality of measurable quantities, and wherein the matrix has a dimension of N×

M, N being the total number of the plurality of cells and M being the total number of the plurality of measurable quantities;

b. exposing at least one of the plurality of cells to an agent;

c. measuring the measurable quantities of the at least one of the plurality of cells responsive to the agent;

d. comparing the measured measurable quantities of the at least one of the plurality of cells responsive to the agent with the corresponding biological signatures of the matrix of biological signatures; and

e. identifying the agent from the comparison.

2. The method of claim 1, further comprising the step of calibrating the matrix of biological signatures from a plurality of chemical agents subsequent to the constructing step.

3. The method of claim 2, further comprising the step of applying preconditioning agents to the plurality of cells to place the plurality of cells in a desired physiological state subsequent to the calibrating step.

4. The method of claim 1, further comprising the step of quantifying an agent from the comparison.

5. The method of claim 2, wherein the plurality of chemical agents comprise toxins.

6. The method of claim 1, wherein the plurality of cells comprises at least one cell line.

7. The method of claim 1, wherein each of the plurality of measurable quantities comprises a physical quantity related to cellular physiological activities of the cell exposed to the agent.

8. The method of claim 1, wherein the measuring step comprises the steps of:

a. measuring a first biology signature for a first cell responsive to the agent, wherein the first biology signature for a first cell corresponds to a first element of the matrix in a first row;

b. measuring a second biology signature for the first cell responsive to the agent, wherein the second biology signature corresponds to an element next to the first element of the matrix in the first row; and

c. repeating step (b) until all biology signatures corresponding to the elements in the first row of the matrix have been measured.

9. The method of claim 8, wherein the measuring step further comprises the steps of:

a. measuring a first biology signature for a second cell responsive to the agent, wherein the first biology signature for a second cell corresponds to a first element of the matrix in a second row that is next to the first row;

b. measuring a second biology signature for the second cell responsive to the agent, wherein the second biology signature corresponds to an element next to the first element of the matrix in the second row;

c. repeating step (b) until all biology signatures corresponding to the elements in the second row of the matrix have been measured; and

d. repeating steps (a)-(c) but treating the second row as the first row until all biology signatures corresponding to the elements in all N rows of the matrix have been measured.

10. The method of claim 9, wherein the biology signatures corresponding to the elements in all N rows of the matrix can be measured simultaneously.

11. The method of claim 9, wherein the biology signatures corresponding to the elements in all N rows of the matrix can be measured in sequence.

12. The method of claim 1, further comprising the steps of:

a. keeping the exposure of the at least one of the plurality of cells to the agent under a threshold of exposure for irreversible cell damage or cell death; and

b. adjusting the exposure of the at least one of the plurality of cells to the agent according to the measured measurable quantities.

13. The method of claim 1, further comprising the step of storing the measured measurable quantities.

14. The method of claim 1, wherein the agent comprises a chemical agent.

15. The method of claim 1, wherein the agent comprises a biological agent.

16. An apparatus for using biological material to discriminate an agent, comprising:

a. means for constructing a matrix of biological signatures having a plurality of elements, wherein each element represents a biological signature of one of a plurality of cells in response to an agent, each biological signature being one of a plurality of measurable quantities, and wherein the matrix has a dimension of N×

M, N being the total number of the plurality of cells and M being the total number of the plurality of measurable quantities;

b. means for exposing at least one of the plurality of cells to an agent;

c. means for measuring the measurable quantities of the at least one of the plurality of cells responsive to the agent;

d. means for comparing the measured measurable quantities of the at least one of the plurality of cells responsive to the agent with the corresponding biological signatures of the matrix of biological signatures; and

e. means for identifying the agent from the comparison.

17. The apparatus of claim 16, wherein the exposing means comprises a chamber for receiving the at least one of the plurality of cells.

18. The apparatus of claim 16, wherein the measuring means comprises a sensor.

19. The apparatus of claim 16, wherein the comparing means comprises a controller.

20. The apparatus of claim 16, further comprising means for calibrating the matrix of biological signatures from a plurality of chemical agents.

21. The apparatus of claim 16, further comprising means for storing the measured measurable quantities.

22. The apparatus of claim 21, wherein the storing means comprises a memory device.

23. The apparatus of claim 16, further comprising means for quantifying an agent from the comparison.

24. The apparatus of claim 16, wherein the agent comprises a chemical agent.

25. The apparatus of claim 16, wherein the agent comprises a biological agent.

26. A method for using biological material to discriminate an agent, comprising the steps of:

a. providing at least one cell;

b. exposing at least one cell to an agent;

c. measuring the response of the cell to the agent in terms of a physical quantity related to at least one of the cellular physiological activities of the cell; and

d. identifying the agent from the measured response.

27. The method of claim 26, further comprising the step of quantifying the agent from the measured response.

28. An apparatus for using biological material to discriminate an agent, comprising:

a. means for providing at least one cell;

b. means for exposing at least one cell to an agent;

c. means for measuring the response of the cell to the agent in terms of a physical quantity related to at least one of the cellular physiological activities of the cell; and

d. means for identifying the agent from the measured response.

29. The apparatus of claim 28, wherein the exposing means comprises a chamber for receiving the at least one cell.

30. The apparatus of claim 28, wherein the measuring means comprises a sensor.

31. The apparatus of claim 28, wherein the identifying means comprises a controller.

32. The apparatus of claim 28, further comprising means for quantifying the agent from the measured response.

33. A method for using biological material to discriminate an agent, comprising the steps of:

a. providing at least one cell;

b. exposing at least one cell to an agent;

c. measuring the response of the cell to the agent, where the response of the cell to the agent is characterized by a reaction time, at a time period shorter than the reaction time; and

d. identifying the agent from the measured response.

34. The method of claim 33, further comprising the step of quantifying the agent from the measured response.

35. The method of claim 33, wherein the response comprises a temporal response and the temporal response of the cell to the agent is measured in at least two measurements.

36. The method of claim 33, wherein the time between the measurements is shorter than the reaction time corresponding to the temporal response of the cell.

37. An apparatus for using biological material to discriminate an agent, comprising:

a. means for providing at least one cell;

b. means for exposing at least one cell to an agent;

c. means for measuring the response of the cell to the agent, where the response of the cell to the agent is characterized by a reaction time, at a time period shorter than the reaction time; and

d. means for identifying the agent from the measured response.

38. The apparatus of claim 37, wherein the exposing means comprises a chamber for receiving the at least one cell.

39. The apparatus of claim 37, wherein the measuring means comprises a sensor.

40. The apparatus of claim 37, wherein the identifying means comprises a controller.

41. The method of claim 37, further comprising means for quantifying the agent from the measured response.

42. The method of claim 37, wherein the response comprises a temporal response and the temporal response of the cell to the agent is measured in at least two measurements.

43. The method of claim 37, wherein the time between the measurements is shorter than the reaction time corresponding to the temporal response of the cell.

44. A method for using biological material to discriminate an agent, comprising the steps of:

a. providing a plurality of cells in a conditioned environment, each cell having multiple metabolic pathways for metabolic events;

b. simultaneously monitoring of multiple metabolic pathways for each of the plurality of cells, wherein each metabolic pathway may be disturbed in the presence of an agent;

c. detecting the changes of metabolic events for each of the plurality of cells at a time when an agent invades the conditioned environment; and

d. identifying the agent from the detected changes of metabolic events for each of the plurality of cells within seconds from the time when the agent invades the conditioned environment.

45. The method of claim 44, wherein each of the metabolic events involves a change in energy production or consumption of the cell.

46. The method of claim 44, further comprising the step of quantifying the agent from the measured response.

55. The method of claim 44, wherein each of the metabolic events involves a change in energy production or consumption of the cell.

56. The method of claim 44, further comprising the step of quantifying the agent from the measured response.

47. An apparatus for using biological material to discriminate an agent, comprising:

a. means for providing a plurality of cells in a conditioned environment, each cell having multiple metabolic pathways for metabolic events;

b. means for simultaneously monitoring of multiple metabolic pathways for each of the plurality of cells, wherein each metabolic pathway may be disturbed in the presence of an agent;

c. means for detecting the changes of metabolic events for each of the plurality of cells at a time when an agent invades the conditioned environment; and

d. means for identifying the agent from the detected changes of metabolic events for each of the plurality of cells within seconds from the time when the agent invades the conditioned environment.

48. The apparatus of claim 47, wherein the providing means comprises at least one chamber for receiving the plurality of cells in the conditioned environment.

49. The apparatus of claim 47, wherein the monitoring means comprises a sensor.

50. The apparatus of claim 47, wherein the identifying means comprises a controller.

51. The apparatus of claim 47, further comprising means for quantifying the agent from the measured response.

52. An apparatus for using biological material to discriminate an agent, comprising:

a. a plurality of cells, each cell having multiple metabolic pathways for metabolic events;

b. a structure for receiving the plurality of cells, the structure having a plurality of sensing volumes, each sensing volume being in a conditioned environment capable of receiving at least one cell;

c. an array of sensors positioned underneath the structure for simultaneously monitoring of multiple metabolic pathways for each of the plurality of cells, wherein each metabolic pathway may be disturbed in the presence of an agent; and

d. a controller coupled to each sensor of the array of the sensors, wherein when an agent invades the conditioned environment, the array of the sensors detects the changes of metabolic events for at least one of the cells and generates at least one signal in response, and wherein the controller receives the signal from the array of the sensors and identifies the agent from the signal.

53. The apparatus of claim 52, wherein the controller further quantifies the agent from the signal.

54. A method for using biological material to discriminate an agent, comprising the steps of:

a. providing a plurality of cells in a conditioned environment, each cell having multiple metabolic pathways for metabolic events;

b. simultaneously monitoring of multiple metabolic pathways for each of the plurality of cells, wherein each metabolic pathway may be disturbed in the presence of an agent;

c. detecting the changes of metabolic events for each of the plurality of cells at a time when an agent invades the conditioned environment; and

d. identifying the agent from the detected changes of metabolic events for each of the plurality of cells within seconds from the time when the agent invades the conditioned environment.

57. An apparatus for using biological material to discriminate an agent, comprising:

a. means for providing a plurality of cells in a conditioned environment, each cell having multiple intracellular and extracellular signalling pathways for intracellular and extracellular signalling events;

b. means for simultaneously monitoring of multiple intracellular and extracellular signalling pathways for each of the plurality of cells, wherein each intracellular and extracellular signalling pathway may be disturbed in the presence of an agent;

c. means for detecting the changes of intracellular and extracellular signalling events for each of the plurality of cells at a time when an agent invades the conditioned environment; and

d. means for identifying the agent from the detected changes of intracellular and extracellular signalling events for each of the plurality of cells within seconds from the time when the agent invades the conditioned environment.

58. The apparatus of claim 57, wherein the providing means comprises at least one chamber for receiving the plurality of cells in the conditioned environment.

59. The apparatus of claim 57, wherein the monitoring means comprises a sensor.

60. The apparatus of claim 57, wherein the identifying means comprises a controller.

61. The apparatus of claim 57, further comprising means for quantifying the agent from the measured response.

62. An apparatus for using biological material to discriminate an agent, comprising:

a. a plurality of cells, each cell having multiple intracellular and extracellular signalling pathways for intracellular and extracellular signalling events;

b. a structure for receiving the plurality of cells, the structure having a plurality of sensing volumes, each sensing volume being in a conditioned environment capable of receiving at least one cell;

c. an array of sensors positioned underneath the structure for simultaneously monitoring of multiple intracellular and extracellular signalling pathways for each of the plurality of cells, wherein each intracellular and extracellular signalling pathway may be disturbed in the presence of an agent; and

d. a controller coupled to each sensor of the array of the sensors, wherein when an agent invades the conditioned environment, the array of the sensors detects the changes of intracellular and extracellular signalling events for at least one of the cells and generates at least one signal in response, and wherein the controller receives the signal from the array of the sensors and identifies the agent from the signal.

63. The apparatus of claim 62, further comprising means for quantifying the agent from the measured response.