Method and apparatus for UV imaging

• US 7,491,943 B2
• Filed: 01/13/2006
• Issued: 02/17/2009
• Est. Priority Date: 01/13/2005
• Status: Expired due to Fees

• Current Assignee: Massachusetts Institute of Technology, Whitehead Institute for Biomedical Research
• Original Assignee: Massachusetts Institute of Technology, Whitehead Institute for Biomedical Research
• Inventors: Zeskind, Benjamin J., Ehrlich, Daniel, Matsudaira, Paul
• Primary Examiner(s): Porta; David P
• Assistant Examiner(s): Malevic; Djura
• Application Number:
  US11/331,982
• Publication Number:
  US 20060188869A1
• Time in Patent Office: 1,131 Days
• Field of Search: 250/461.1, 250/461.2, 250/459.1, 250/458.1
• US Class Current: 250/372
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 21/19 Dichroism
• CPC Class Codes: G01N 21/33 using ultraviolet light G01N21/39 takes precedence
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging
• 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: G02B 21/16 adapted for ultraviolet illumination ; Fluorescence microscopes G02B21/0076 takes precedence

Abstract

A method and apparatus for illuminating a subject with deep UV light and detecting 2-dimensional image information, e.g., a transmission image, fluorescence image, reflected light image, etc. The subject may be illuminated with a light pulse having a duration of 2 seconds or less and one or more components with a wavelength of between about 150 nm to about 300 nm. The light pulse may be arranged so as to minimize or eliminate any detrimental effect on the functioning of the subject, e.g., on a living cell. A concentration, mass, quantum yield, absorbance or other characteristics of one or more compounds in the subject may be determined based on one or more 2-dimensional images obtained of the subject. An illumination beam having left and right circularly polarized components may be generated, e.g., for circular dichroism analysis of a subject.

Claims

1. A method for imaging a subject, comprising:

providing a subject;

illuminating the subject with a light pulse having a wavelength of between about 150 nm to about 300 nm, the light pulse having a time duration of about 2 seconds or less;

detecting illumination that results from interaction of the light pulse with the subject to capture 2-dimensional image information for at least a portion of the subject; and

waiting a time interval of at least 5 seconds before repeating performances of the illuminating and detecting steps to capture additional 2-dimensional image information for at least a portion of the subject.

2. The method of claim 1, wherein the step of providing a subject comprises providing a living subject, and wherein the step of illuminating the subject does not result in significant effect on functioning of the living subject.

3. The method of claim 2, wherein the steps of illuminating and detecting are repeated multiple times with the living subject without causing significant effect on functioning of the living subject by repeating the steps of illuminating and detecting multiple times.

4. The method of claim 3, wherein time duration of the light pulse is less than 1 second.

5. The method of claim 3, wherein the time duration of the light pulse is about 100 milliseconds and the wavelength of the light pulse is about 280 nm.

6. The method of claim 1, wherein the step of detecting is performed using an electronic imaging device adapted to detect image information in 2 dimensions.

7. The method of claim 6, wherein a timing of emission of the light pulse is synchronized with operation of the electronic imaging device.

8. The method of claim 1, wherein the steps of illuminating and detecting are performed using brightfield illumination of the subject to capture a transmission image of the subject.

9. The method of claim 1, wherein the subject includes a living cell or a biological material other than artificially crystallized proteins.

10. An apparatus for imaging a subject, comprising:

an illumination system that illuminates the subject with a light pulse having a wavelength of about 150 nm to about 300 nm, the light pulse having a time duration of about 2 seconds or less, the illumination system constructed and arranged to prevent a waiting time interval of less than 5 seconds between performances of illuminating the subject with the light pulse to capture 2-dimensional image information for at least a portion of the subject; and

a detector tat detects illumination that results from interaction of the light pulse with the subject so as to capture 2-dimensional image information of at least a portion of the subject.

11. The apparatus of claim 10, wherein the subject comprises a living subject, and wherein the illumination system is adapted to illuminate the living subject without causing significant effect on functioning of the living subject.

12. The apparatus of claim 11, wherein the illumination system is adapted to illuminate the living subject with a light pulse multiple times without causing significant effect on functioning of the living subject after illumination with multiple light pulses.

13. The apparatus of claim 12, wherein the time duration of each light pulse is about 100 milliseconds and the wavelength of the light pulse is about 280 mn.

14. The apparatus of claim 10, wherein the detector includes an electronic imaging device adapted to detect image information in 2 dimensions, and wherein a timing of emission of the light pulse is synchronized with operation of the electronic imaging device.

15. The apparatus of claim 10, wherein the detector detects light from the light pulse that is transmitted through the subject.

16. The apparatus of claim 10, wherein the detector is adapted to obtain multiple 2-dimensional images to provide 3-dimensional image information regarding the subject.

17. A method for analyzing a subject, comprising:

providing a subject including at least first and second different compounds, said compounds being absorbent of light in a wavelength range of between about 150 mn to about 300 mn;

illuminating the subject with light in the wavelength range for a first time;

detecting illumination of a first wavelength or range of wavelengths that results from interaction of the light in the wavelength range with the subject so as to capture first 2-dimensional image information of at least a portion of the subject;

illuminating the subject with light in the wavelength range for a second time after waiting a time interval of at least 5 seconds after performance of illuminating the subject for the first time and detecting illumination of the first wavelength or range or wavelengths;

detecting illumination of a second wavelength or range of wavelengths that results from interaction of the light in the wavelength range with the subject so as to capture second 2-dimensional image information of at least a portion of the subject; and

using the first and second 2-dimensional image information to determine concentrations of the first and second compounds in the subject.

18. The method of claim 17, wherein the step of using the first and second 2-dimensional image information comprises determining relative concentrations of the first and second compounds for a plurality of pixel or voxel locations.

19. The method of claim 17, wherein the step of using the first and second 2-dimensional image information comprises determining absolute concentrations or masses of the first and second compounds for a plurality of pixel or voxel locations.

20. The method of claim 17, wherein the step of using the first and second 2-dimensional image information comprises determining a contribution of each of the first and second compounds to an optical density for each of a plurality of pixel or voxel locations.

21. The method of claim 17, wherein the first and second compounds are protein and nucleic acid, respectively.

22. The method of claim 17, wherein the steps of illuminating comprise illuminating the subject with a brightfield pulse of light having a wavelength below 300 nm.

23. The method of claim 17, wherein the steps of illuminating comprise illuminating the subject with a brightfield pulse of light having a time duration of less than 2 seconds.

24. The method of claim 17, wherein the subject includes a living cell, and the steps of illuminating comprise illuminating the subject with a brightfield pulse of light having a wavelength below 300 nm such that the brightfield pulse of light does not cause significant effect on the functioning of the living cell.

25. The method of claim 17, further comprising generating information for visible display as an image of the concentration of the first and second compounds.

26. A method for imaging a subject, comprising:

providing a subject including a living cell and/or a biological material other than artificially crystallized proteins;

illuminating the subject with illumination having a wavelength of about 300 nm or less;

detecting autofluorescent illumination emitted by the subject that results from interaction of the illumination with the subject so as to capture 2-dimensional image information of at least a portion of the subject; and

waiting a time interval of at least 5 seconds before repeating performances of the illuminating and detecting steps to capture additional 2-dimensional image information for at least a portion of the subject.

27. The method of claim 26, wherein the steps of illuminating and detecting are performed using a microscopy and/or endoscopy device.

28. The method of claim 26, wherein the step of illuminating the subject comprises illuminating the subject using a diafluorescence illumination arrangement.

29. A method for imaging a subject, comprising:

providing a subject;

illuminating the subject a first time with illumination having a wavelength of about 350 nm or less, the illumination being left or right circularly polarized;

detecting illumination that results from interaction of the left or right circularly polarized illumination with the subject so as to capture first 2-dimensional image information of at least a portion of the subject;

illuminating the subject a second time with illumination having a wavelength of about 350 nm or less after waiting a time interval of at least 5 seconds after illuminating the subject the first time, the illumination of the subject the second time being the other of left or right circularly polarized; and

detecting illumination that results from interaction of the other of left or right circularly polarized illumination with the subject so as to capture second 2-dimensional image information of at least a portion of the subject.

30. The method of claim 29, wherein the steps of illuminating and detecting are performed using a microscopy and/or endoscopy device.

31. The method of claim 29, wherein the step of illuminating the subject comprises illuminating the subject using a diafluorescence illumination arrangement.

32. The method of claim 29, wherein the left and right circularly polarized illumination has a wavelength suitable to cause at least one compound in the subject to autofluoresce;

and wherein the steps of detecting comprise detecting autofluorescent illumination that is emitted by the at least one compound in response to the left and right circularly polarized illumination.