Methods and systems for assessing biological materials using optical and spectroscopic detection techniques

• US 6,319,682 B1
• Filed: 07/31/2000
• Issued: 11/20/2001
• Est. Priority Date: 10/04/1995
• Status: Expired due to Fees

• Current Assignee: Neurotherapeutics Pharma LLC
• Original Assignee: Cytoscan Sciences, LLC
• Inventors: Hochman, Daryl W.
• Primary Examiner(s): Leary, Louise N.
• Application Number:
  US09/629,046
• Time in Patent Office: 477 Days
• Field of Search: 435/29, 435/4, 435/30, 435/288.7, 435/32
• US Class Current: 435/29
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: C12Q 1/02 involving viable microorganisms
• 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 33/6896 Neurological disorders, e.g. Alzheimer's disease

Abstract

Optical detection techniques for the assessment of the physiological state, health and/or viability of biological materials are provided. Biological materials which may be examined using such techniques include cells, tissues, organs and subcellular components. The inventive techniques may be employed in high throughput screening of potential diagnostic and/or therapeutic agents.

Claims

1. A method for screening a candidate compound for activity as a chloride cotransporter agonist or antagonist, comprising:

introducing an agent to a cell sample that is capable of changing at least one of the ionic concentration and the ionic balance in a cell sample without killing cells;

observing and recording at least one property of the cell sample that is representative of at least one of the ionic concentration and the ionic balance in the cell sample following introduction of the agent;

introducing a candidate compound to the cell sample;

observing and recording at least one property of the cell sample that is representative of at least one of the ionic concentration and the ionic balance in the cell sample following introduction of the candidate compound; and

comparing the at least one property of the cell sample recorded following introduction of the agent with the at least one property of the cell sample recorded following introduction of the candidate compound, and thereby determining changes in at least one of the ionic concentration and the ionic balance in the cell sample.

2. A method of claim 1, wherein the at least one property is a geometrical or optical property of the cell sample.

3. A method of claim 1, wherein the at least one property is an optical property selected from the group consisting of:

light scattering, reflectance, absorption, refraction, diffraction, birefringence, refractive index or Kerr effect.

4. A method of claim 1, wherein the at least one property is a geometrical property selected from the group consisting of:

cell diameter, cell volume, cell confirmation, intracellular space, or extracellular space.

39. A method of any of claims 1 or 5 or 19 or 31, wherein the at least one sample population of biological materials is maintained in a cell culture system.

40. A method of any of claims 1 or 5 or 19 or 31, wherein the at least one sample population of biological materials is maintained in a tissue culture system.

41. A method of any of claims 1 or 5 or 19 or 31, wherein the at least one sample population of biological materials is maintained in an organ culture system.

42. A method of any of claims 1 or 5 or 19 or 31, wherein the at least one sample population of biological materials is maintained in an organism.

43. A method of any of claims 1 or 5 or 19 or 31, wherein the at least one sample population of biological materials comprises genetically modified cells or tissues.

44. A method of any of claims 1 or 5 or 19 or 31, wherein the at least one sample population of biological materials comprises transfected cells or tissues.

45. A method of any of claims 1 or 5 or 19 or 31, wherein the at least one sample population of biological materials comprises cells or tissue of a transgenic a or plant.

5. A method for assessing the response of a sample biological population to a challenge comprising:

maintaining at least one sample population of biological material in a viable condition;

administering a challenge to the sample population;

acquiring a test data set corresponding to one or more geometrical or optical properties of the sample population following the challenge;

comparing the test data set to a comparison data set, wherein the comparison data set corresponds to the one or more geometrical or optical properties of a predetermined sample population having a predetermined condition; and

assessing the response of the sample population to the challenge based on the comparison.

6. A method of claim 5, wherein the comparison data set is acquired by determining the one or more geometrical or optical properties of the sample population prior to the challenge.

7. A method of claim 5, wherein the comparison data set is acquired by determining the one or more geometrical or optical properties of a control sample population having a similar population and physiological state as the sample population prior to the challenge.

8. A method of claim 5, additionally comprising acquiring multiple comparison data sets corresponding to the one or more geometrical or optical properties of the sample population at different times after administration of the challenge.

9. A method of claim 5, wherein the challenge produces a change in electrical activity in the sample population.

10. A method of claim 5, wherein the sample population is in a viable, healthy condition prior to administration of the challenge.

11. A method of claim 5, wherein the challenge comprises an agent selected from the group consisting of:

pathogens, pollutants;

radiation;

chemotherapeutic agents;

inmumodulatory agents;

biological agents;

infectious agents;

apostolic agents;

necrotic agents;

mechanical insult;

stress;

aging agents;

or disease agents.

12. A method of claim 5, wherein the challenge is a physiological stimuli.

13. A method of claim 5, wherein the challenge produces a change in at least one of the physiological characteristics of the sample population selected from the group consisting of:

extracellular osmolarity, ion concentration, oxygen condition, nutrient condition, metabolite condition, ion homeostasis, electrical stimulations electrical inhibition, synchrony, or extracellular space.

14. A method of claim 5, wherein the challenge produces a genetic modification in the sample population.

15. A method of claim 5, wherein the challenge produces a change in gene expression in the sample population.

16. A method of claim 5, additionally comprising simulating a disease state in the sample population prior to administration of the challenge.

17. A method of claim 5, wherein the sample population is in a dysfunctional condition prior to administration of the challenge.

18. A method of claim 5, wherein the sample population is in a malignant condition prior to administration of the challenge.

19. A method for screening a candidate compound for activity as a diagnostic or therapeutic agent, comprising:

maintaining at least one sample population of biological material;

introducing a candidate compound to the sample population;

acquiring and recording at least one test data set corresponding to one or more geometrical or optical properties of the sample population following introduction of the candidate compound;

comparing the test data set to a comparison data set, wherein the comparison data set corresponds to one or more geometrical or optical properties of a predetermined sample population having a predetermined physiological state; and

assessing the activity of the candidate compound as a diagnostic or therapeutic agent based on the comparison between the test data set and the comparison data set.

20. A method according to claim 19, wherein the candidate compound is screened as an agent for diagnosis or treatment of a condition selected from the group consisting of:

migraine headache, spreading depression, epilepsy, Alzheimer'"'"'s disease, multiple sclerosis, psychiatric disorders, depression, anxiety, bipolar disorder, schizophrenia, Parkinson'"'"'s disease, inflammation, trauma, mechanical injury, anoxia, stroke, ischemia, hypoxia, malignancies, angiogenesis, wound healing, and immune deficiencies.

21. A method of claim 19, wherein the comparison data set is acquired by determining the one or more geometrical or optical properties of the sample population prior to introduction of the candidate compound.

22. A method of claim 19, wherein the comparison data set is acquired by determining the one or more geometrical or optical properties of a control sample population having a similar population and physiological state as the sample population prior to introduction of the candidate compound.

23. A method of claim 19, additionally comprising acquiring multiple comparison data sets corresponding to the one or more geometrical or optical properties of the sample population at different times after introduction of the candidate compound.

24. A method of claim 19, wherein the sample population is in a viable, healthy condition prior to introduction of the candidate compound.

25. A method of claim 19, wherein the candidate compound comprises an agent selected from the group consisting of:

pathogens, pollutants;

radiation;

chemotherapeutic agents;

inmumodulatory agents;

biological agents;

infectious agents;

apostolic agents;

necrotic agents;

mechanical insult;

stress;

aging agents;

or disease agents.

26. A method of claim 19, wherein the candidate compound produces a change in at least one of the physiological characteristics of the sample population selected from the group consisting of:

extracellular osmolarity, ion concentration, oxygen condition, nutrient condition, metabolite condition, ion homeostasis, electrical stimulation, electrical inhibition, synchrony, or extracellular space.

27. A method of claim 19, wherein the candidate compound produces a genetic modification in the sample population.

28. A method of claim 19, additionally comprising simulating a disease state in the sample population prior to introduction of the candidate compound.

29. A method of claim 19, wherein the sample population is in a dysfunctional condition prior to introduction of the candidate compound.

30. A method of claim 19, wherein the sample population is in a malignant condition prior to introduction of the candidate compound.

33. A method of any of claims 5 or 19 or 31, wherein the comparison data set is an empirically determined standard for a predetermined sample population type and physiological state.

34. A method of any of claims 5 or 19 or 31, wherein the sample population is part of an animal or a plant and the assessment is done on an in situ sample population.

35. A method of any of claims 5 or 19 or 31, wherein the sample population is a cell or tissue or organ culture maintained in vitro.

36. A method of any of claims 5 or 19 or 31, wherein test data corresponding to one or more optical properties is acquired and the one or more optical properties are selected from the group consisting of:

light scattering, reflection, absorption, refraction, diffraction, birefringence, refractive index, or Kerr effect.

37. A method of any of claims 5 or 19 or 31, wherein test data corresponding to one or more geometrical properties of the sample population is acquired and the one or more geometrical properties are selected from the group consisting of:

cell diameter;

cell volume;

cell confirmation;

intracellular space or extracellular space.

38. A method of any of claims 5 or 19 or 31, additionally comprising introducing a contrast enhancing agent to the sample population prior to acquisition of the test data set.

46. A method according to any of claims 5 or 19 or 31, additionally comprising illuminating the sample population with an electromagnetic radiation (emr) source prior to acquisition of the test data set, wherein the emr source is selected from the group consisting of:

high intensity emr sources, broad spectrum emr sources, non-chromatic emr sources, tungsten-halogen lamps, lasers, light emitting diodes, laser diodes, polarized emr sources and dark field illumination sources.

47. A method according to any of claims 5 or 19 or 31, wherein the sample population is illuminated with emr delivered to the sample population through an optical fiber.

48. A method according to any of claims 5 or 19 or 31, wherein the test data is acquired by an optical detector selected from the group consisting of:

cameras, photo diodes, photo multiplier tubes, photo sensitive silicon detector chips, or CCD devices.

49. A method according to any of claims 5 or 19 or 31, additionally comprising acquiring a series of data sets representing one or more geometrical or optical properties and combining the series of data sets to obtain a data profile.

50. A method according to any of claims 5 or 19 or 31, additionally comprising magnifying the sample population sufficiently to discern single cells in the sample population.

51. A method according to any of claims 5 or 19 or 31, wherein the at least one sample population of biological materials is maintained in a culture system and the sample population is illuminated continuously during acquisition of test data.

52. A method according to any of claims 5 or 19 or 31, wherein the at least one sample population of biological materials is maintained in a culture system and the sample population is illuminated non-continuously during acquisition of test data.

53. A method according to any of claims 5 or 19 or 31, wherein the at least one sample population of biological materials is maintained in a culture system and the sample population is illuminated using a technique selected from the group consisting of:

amplitude modulated, frequency modulated or phase modulated techniques.

31. A method for assessing the viability of a biological sample, comprising:

acquiring test data corresponding to one or more geometrical or optical properties of the sample population;

comparing the test data to comparison data corresponding to the one or more geometrical or optical properties of a population having predetermined viability properties; and

assessing the viability of the sample population based on the comparison.

32. A method of claim 31, additionally comprising classifying the sample population as viable or non-viable.