Wide field method for detecting pathogenic microorganisms

• US 7,057,721 B2
• Filed: 06/27/2003
• Issued: 06/06/2006
• Est. Priority Date: 01/10/2002
• Status: Active Grant

• Current Assignee: Chemimage Corporation
• Original Assignee: Chemimage Corporation
• Inventors: Nelson, Matthew P., Treado, Patrick J., Schweitzer, Robert C., Vanni, G. Steven, Wolfe, Julianne, Gardner, Charles W. Jr., Maier, John S.
• Primary Examiner(s): EVANS, FANNIE L
• Application Number:
  US10/608,470
• Publication Number:
  US 20050185178A1
• Time in Patent Office: 1,075 Days
• Field of Search: 356/301
• US Class Current: 356/301
• CPC Class Codes: G01J 3/02 Details
• CPC Class Codes: G01J 3/0272 Handheld
• CPC Class Codes: G01J 3/10 Arrangements of light sources specially adapted for spectrometry or colorimetry
• CPC Class Codes: G01J 3/44 Raman spectrometry; Scattering spectrometry ; Fluorescence spectrometry
• CPC Class Codes: G01N 2021/6417 Spectrofluorimetric devices
• CPC Class Codes: G01N 2021/6423 Spectral mapping, video display
• CPC Class Codes: G01N 2021/656 Raman microprobe
• CPC Class Codes: G01N 21/35 using infrared light G01N21/39 takes precedence
• CPC Class Codes: G01N 21/3577 for analysing liquids, e.g. polluted water
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 21/65 Raman scattering
• CPC Class Codes: G01N 2201/0221 Portable; cableless; compact; hand-held

Abstract

Pathogenic microorganisms are detected in a wide field of view and classified by Raman light scattered light from these organisms together with digital pattern recognition of their spectral patterns.

Claims

1. A method for investigating the presence of Anthrax (Bacillus anthracis) on a substrate, comprising:

a) selecting a first area from the surface of the substrate, the first area sufficiently large to contain a small plurality of Anthrax organisms, then

b) selecting a wide field of view within the first area for further investigation, then

c) irradiating the wide field of view plus an area surrounding the wide field of view with sufficient energy to photobleach the irradiated area, then

d) collecting Raman shifted light from the wide field of view, then

e) analyzing the Raman shifted light for patterns characteristic of Anthrax microorganisms.

2. The method of claim 1, further comprising repeating steps a) to e) for a second area.

3. The method of claim 1, where the step of analyzing includes analyzing the strain of the Anthrax microorganisms.

4. The method of claim 1, where the step of analyzing includes analyzing the viability of the Anthrax microorganisms.

5. The method of claim 1, where the step of analyzing includes analyzing the growth medium in which the Anthrax microorganisms has been grown.

6. A method for investigating the presence of pathogenic microorganisms on a substrate, comprising:

a) selecting a first area from the surface of the substrate, the first area sufficiently large to contain a small plurality of pathogenic microorganisms, then

b) selecting a wide field of view within the first area for further investigation, then

c) irradiating the wide field of view plus an area surrounding the wide field of view with sufficient energy to photobleach the irradiated area, then

d) collecting Raman shifted light from the wide field of view, then

e) analyzing the Raman shifted light for patterns characteristic of pathogenic microorganisms.

7. The method of claim 6, further comprising repeating steps a) to e) for a second area.

8. The method of claim 6, where the step of analyzing includes analyzing the growth medium in which the pathogenic microorganisms has been grown.

9. The method of claim 6, where the Raman shifted light from the wide field passes thru a spot, line or image focusing type of spectrometer.

10. The method of claim 6, where the pathogenic microorganisms are anthrax spores.

11. The method of claim 6, where the pathogenic microorganisms are protozoa.

12. The method of claim 6, where the pathogenic microorganisms are cryptosporidia microorganisms.

13. The method of claim 6, where the pathogenic microorganisms are Escherichia coli microorganisms.

14. The method of claim 6, where the pathogenic microorganisms are Escherichia coli 157 microorganisms.

15. The method of claim 6, where the pathogenic microorganisms are Plague (Yersinia pestis) microorganisms.

16. The method of claim 6, where the pathogenic microorganisms are Smallpox (variola ajor) microorganisms.

17. The method of claim 6, where the pathogenic microorganisms are Tularemia (Francisella tularensis) microorganisms.

18. The method of claim 6, where the pathogenic microorganisms are Brucellosis (Brucella species) microorganisms.

19. The method of claim 6, where the pathogenic microorganisms are Clostridium perfringens microorganisms.

20. The method of claim 6, where the pathogenic microorganisms are Salmonella microorganisms.

21. The method of claim 6, where the pathogenic microorganisms are Shigella microorganisms.

22. The method of claim 6, where the pathogenic microorganisms are Glanders (Burkholderia mallei) microorganisms.

23. The method of claim 6, where the pathogenic microorganisms are Melioidosis (Burkholderia pseudomallei) microorganisms.

24. The method of claim 6, where the pathogenic microorganisms are Psittacosis Chlamydia psittci) microorganisms.

25. The method of claim 6, where the pathogenic microorganisms are Q fever Coxiella burnetii) microorganisms.

26. The method of claim 6, where the pathogenic microorganisms are Typhus ever (Rickettsia prowazekii) microorganisms.

27. The method of claim 6, where the pathogenic microorganisms are Vibrio cholerae microorganisms.

28. The method of claim 6, where the pathogenic microorganisms are chosen from the group of filoviruses (such as Ebola and Marburg viruses), naviruses (such as Lassa fever and Machupo viruses) and alphaviruses (such as Venezuelan equine encephalitis, eastern equine encephalitis, and western equine encephalitis).

29. The method of claim 6, where the pathogenic microorganisms are Giardia microorganisms.

30. The method of claim 6, where the pathogenic microorganisms are Candida albicans microorganisms.

31. The method of claim 6, where the pathogenic microorganisms are Enterococcus faecalis microorganisms.

32. The method of claim 6, where the pathogenic microorganisms are Staphylococcus epidermidis microorganisms.

33. The method of claim 6, where the pathogenic microorganisms are Staphylococcus aureus microorganisms.

34. The method of claim 6, where the pathogenic microorganisms are Enterobacter aerogenes microorganisms.

35. The method of claim 6, where the pathogenic microorganisms are Corynebacterium diphtheriae micorganisms.

36. The method of claim 6, where the pathogenic microorganisms are Pseudomonas aeruginosa microorganisms.

37. The method of claim 6, where the pathogenic microorganisms are Acinetobacter calcoaceticus microorganisms.

38. The method of claim 6, where the pathogenic microorganisms are Klebsiella pneumoniae microorganisms.

39. The method of claim 6, where the pathogenic microorganisms are Serratia marcescens microorganisms.

40. The method of claim 6, where the irradiating light is in the ultraviolet spectral region with wavelength less than 410 nm.

41. The method of claim 6, where the irradiating light is in the visible spectral region with wavelength less than 780 nm and greater than 410 nm.

42. The method of claim 6, where the irradiating light is in the near infrared spectral region with wavelength less than 2500 nm and greater than 780 nm.

43. The method of claim 6, where the step of analyzing includes analyzing the strain of the pathogenic microorganisms.

44. The method of claim 6, where the step of analyzing includes analyzing the viability of the pathogenic microorganisms.

45. The method of claim 6, where miniaturized components are used to enable a handheld detector.