Laser apparatus and method for microscopic and spectroscopic analysis and processing of biological cells

• US 5,608,519 A
• Filed: 03/20/1995
• Issued: 03/04/1997
• Est. Priority Date: 03/20/1995
• Status: Expired due to Term

• Current Assignee: Paul L. Gourley
• Original Assignee: Mark F. Gourley, Paul L. Gourley
• Inventors: Gourley, Paul L., Gourley, Mark F.
• Primary Examiner(s): HANTIS, KAREN
• Application Number:
  US08/407,345
• Time in Patent Office: 715 Days
• Field of Search: 356/318, 356/301, 356/338-339, 356/246, 356/471, 372/72, 250/461.1, 250/461.2, 250/458.1, 250/459.1
• US Class Current: 356/318
• CPC Class Codes: G01N 15/10 Investigating individual particles
• CPC Class Codes: G01N 15/1429 Signal processing
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/1024 Counting particles by non-optical means
• CPC Class Codes: G01N 2015/1029 Particle size
• CPC Class Codes: G01N 2021/391 Intracavity sample
• CPC Class Codes: G02B 21/32 Micromanipulators structurally combined with microscopes

Abstract

An apparatus and method for microscopic and spectroscopic analysis and processing of biological cells. The apparatus comprises a laser having an analysis region within the laser cavity for containing one or more biological cells to be analyzed. The presence of a cell within the analysis region in superposition with an activated portion of a gain medium of the laser acts to encode information about the cell upon the laser beam, the cell information being recoverable by an analysis means that preferably includes an array photodetector such as a CCD camera and a spectrometer. The apparatus and method may be used to analyze biomedical cells including blood cells and the like, and may include processing means for manipulating, sorting, or eradicating cells after analysis thereof.

Claims

1. Apparatus for analyzing biological cells comprising:

(a) a laser having a resonant optical cavity formed by at least two reflecting mirrors, a gain medium within the resonant optical cavity, and pump means for activating the gain medium; and

(b) an analysis region located within the resonant optical cavity for containing at least one biological cell to be analyzed, the analysis region being disposed along a central portion of the cavity wherein a laser beam is generated by the laser at least partially in response to one of the biological cells present within the analysis region, the lasers beam having information about the biological cell encoded thereupon.

2. The apparatus of claim 1 further including analysis means for receiving a portion of the laser beam and recovering the information about the biological cells.

3. The apparatus of claim 2 in which the information is in the form of an optical characteristic of the laser beam selected from the group consisting of an emission spectrum, a transverse mode profile, an optical intensity, a nonlinear optical signal, a lasing threshold characteristic, or a combination thereof.

4. The apparatus of claim 2 further including processing means for processing of the biological cells.

5. The apparatus of claim 4 in which the processing means includes at least one manipulation laser.

6. The apparatus of claim 1 in which the analysis region is located within a flow cell.

7. The apparatus of claim 6 further including means for supplying the biological cells to the flow cell, and means for accumulating the biological cells after passing through the flow cell.

8. The apparatus of claim 1 in which the gain medium comprises at least in part a fluorescent stain.

9. The apparatus of claim 1 in which the laser beam has an emission wavelength within the wavelength range of about 600 to about 1500 nanometers.

10. The apparatus of claim 1 in which the means for activating the gain medium is a pump laser having a pump beam directed into the gain medium to provide an activated portion thereof.

11. The apparatus of claim 10 further including steering means for steering the pump beam to activate a particular part of the gain medium.

12. The apparatus of claim 1 in which the laser is a semiconductor laser.

13. The apparatus of claim 12 in which the semiconductor laser is a vertical-cavity surface-emitting laser.

14. The apparatus of claim 13 in which one of the reflecting mirrors is a distributed Bragg reflector mirror on a semiconductor substrate, and another reflecting mirror is located on a transparent substrate above the semiconductor substrate.

15. The apparatus of claim 13 in which the pump means for activating the gain medium is a pump laser having a pump beam directed into the gain medium to provide an activated portion thereof.

16. The apparatus of claim 15 further including steering means for steering the pump beam to activate a particular part of the gain medium.

17. The apparatus of claim 13 in which the gain medium includes a semiconductor p-n junction, and the pump means for activating the gain medium is an electrical current.

18. Apparatus for analyzing biological cells comprising:

(a) a first substrate having a lower reflecting mirror thereon, and a gain medium above the lower reflecting mirror;

(b) a second substrate having an upper reflecting mirror formed on a lower surface thereof, the upper and lower reflecting mirrors forming a resonant optical cavity;

(c) an analysis region within the resonant optical cavity for containing at least one biological cell to be analyzed, the analysis region being disposed along a central portion of the cavity; and

(d) pump means for activating the gain medium to generate in combination with one of the cells present within the central portion of the cavity a laser beam having information about the biological cell encoded thereupon.

19. The apparatus of claim 18 in which the lower reflecting mirror is a distributed Bragg reflector mirror.

20. The apparatus of claim 18 in which the pump means for activating the gain medium is a pump laser having a pump beam directed into the gain medium to provide an activated portion thereof.

21. The apparatus of claim 20 further including steering means for steering the pump beam to activate a particular part of the gain medium.

22. The apparatus of claim 18 in which the gain medium includes a semiconductor p-n junction, and the pump means for activating the gain medium is an electrical current.

23. The apparatus of claim 18 further including analysis means for receiving a portion of the laser beam and recovering the information about the biological cells.

24. The apparatus of claim 23 in which the information is in the form of an optical characteristic of the laser beam selected from the group consisting of an emission spectrum, a transverse mode profile, an optical intensity, a nonlinear optical signal, a lasing threshold characteristic, or a combination thereof.

25. The apparatus of claim 23 further including processing means for processing the biological cells.

26. The apparatus of claim 25 in which the processing means includes at least one manipulation laser.

27. The apparatus of claim 18 in which the analysis region is located within a flow cell.

28. The apparatus of claim 27 further including means for supplying the biological cells to the flow cell, and means for accumulating the biological cells after passing through the flow cell.

29. The apparatus of claim 18 in which the gain medium comprises at least in part a fluorescent stain.

30. The apparatus of claim 18 in which the laser beam has an emission wavelength within the wavelength range of about 600 to about 1500 nanometers.

31. A method for analyzing biological cells comprising the steps of:

(a) locating at least one cell to be analyzed within an analysis region within a resonant optical cavity of a laser;

(b) activating a gain medium within the laser by a pump means and generating a laser beam within the cavity of the laser; and

(c) encoding the laser beam with information about at least one of the biological cells present in the analysis region.

32. The method of claim 31 further including the step of recovering the information about the cell by measuring at least one optical characteristic of the laser beam by analysis means.

33. The method of claim 32 in which the optical characteristic is selected from the group consisting of an emission spectrum, a transverse mode profile, an optical intensity, a nonlinear optical signal, a lasing threshold characteristic, or a combination thereof.

34. The method of claim 32 in which the pump means is a pump laser for optically activating the gain medium.

35. The method of claim 34 further including the step of steering a pump beam from the pump laser to activate a particular part of the gain medium for analyzing at least one cell in superposition therewith.

36. The method of claim 32 further including the step of processing a plurality of biological cells after analyzing the cells and recovering information thereabout.

37. The method of claim 36 in which the step of processing the cells includes at least one process selected from the group consisting of sorting, manipulating, or eradicating cells.

38. The method of claim 31 further including the steps of supplying a biological cells to the analysis region located within a flow cell, and accumulating the biological cells after passing through the flow cell.

39. The method of claim 38 further including the step of activating at least one valve for supplying the biological cells to the analysis region.

40. The method of claim 31 further including the step of staining the cells with a fluorescent stain.

41. The method of claim 40 wherein the fluorescent stain comprises at least a part of the gain medium.

42. Apparatus for analyzing biological cells comprising:

(a) a first laser having a resonant optical cavity including a gain medium and an analysis region for containing at least one biological cell to be analyzed; and

(b) a second laser for activating the gain medium to generate a laser beam within the resonant optical cavity of the first laser, with the laser beam having information about the biological cell encoded thereupon.

43. The apparatus of claim 42 in which the first laser is a vertical-cavity surface-emitting laser.