Flow cytometer jet monitor system

• US 5,602,039 A
• Filed: 10/14/1994
• Issued: 02/11/1997
• Est. Priority Date: 10/14/1994
• Status: Expired due to Term

• Current Assignee: University of Washington
• Original Assignee: University of Washington
• Inventors: Van den Engh, Ger
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): FREED, RACHEL
• Application Number:
  US08/323,352
• Time in Patent Office: 851 Days
• Field of Search: 356/72, 356/73, 250/461.2, 209/3.1, 209/3.3, 209/571, 209/576-579, 209/552, 209/127.4, 209/906, 435/2, 435/7.24, 435/29, 436/52, 436/63, 436/164, 422/82.05, 422/82.08
• US Class Current: 436/164
• CPC Class Codes: G01N 15/1404 Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/1406 Control of droplet point

Abstract

A direct jet monitor illuminates the jet of a flow cytometer in a monitor wavelength band which is substantially separate from the substance wavelength band. When a laser is used to cause fluorescence of the substance, it may be appropriate to use an infrared source to illuminate the jet and thus optically monitor the conditions within the jet through a CCD camera or the like. This optical monitoring may be provided to some type of controller or feedback system which automatically changes either the horizontal location of the jet, the point at which droplet separation occurs, or some other condition within the jet in order to maintain optimum conditions. The direct jet monitor may be operated simultaneously with the substance property sensing and analysis system so that continuous monitoring may be achieved without interfering with the substance data gathering and may be configured so as to allow the front of the analysis or free fall area to be unobstructed during processing.

Claims

1. A flow cytometer system which monitors jet conditions comprising:

a. a nozzle container establishing a nozzle volume and having a nozzle exit;

b. a sheath fluid port located within said nozzle volume;

c. a substance introduction port located within said nozzle volume;

d. an analysis area below said nozzle exit wherein a jet occurs;

e. a substance property sensor which senses emissions from said analysis area over a substance wavelength band;

f. a jet illumination source directed towards said analysis area and which emits electromagnetic emissions over a monitor wavelength band which is substantially separate from said substance wavelength band; and

g. a jet condition sensor which receives emissions from said analysis area and which is responsive to said monitor wavelength band.

2. A flow cytometer system which monitors jet conditions as described in claim 1 and further comprising a substance stimulation source directed towards said analysis area.

3. A flow cytometer system which monitors jet conditions as described in claim 2 wherein said substance introduction port introduces a substance and wherein said substance stimulation source comprises an electromagnetic source which causes fluorescence in said substance.

4. A flow cytometer system which monitors jet conditions as described in claim 3 wherein said fluorescence occurs in a fluorescence emission band and wherein said monitor wavelength band is substantially separate from said fluorescence emission band.

5. A flow cytometer system which monitors jet conditions as described in claim 4 wherein said jet illumination source comprises an infrared source.

6. A flow cytometer system which monitors jet conditions as described in claim 5 wherein said jet condition sensor comprises a charge coupled device which is responsive to said infrared source.

7. A flow cytometer system which monitors jet conditions as described in claim 6 wherein said electromagnetic source comprises a laser source.

8. A flow cytometer system which monitors jet conditions as described in claim 7 wherein said analysis area has a front, wherein said infrared source, said charge coupled device, said substance property sensor, and said laser source are configured away from said analysis front.

9. A flow cytometer system which monitors jet conditions as described in claim 1, 3, 5, or 7 and further comprising a controller which is responsive to said jet condition sensor and wherein said controller acts to adjust at least one condition within said jet.

10. A flow cytometer system which monitors jet conditions as described in claim 9 and further comprising a nozzle adjuster connected to said nozzle container and connected to said controller which adjusts the location of said nozzle container.

11. A flow cytometer system which monitors jet conditions as described in claim 1, 3, 5 or 7 and further comprising an oscillator to which said sheath fluid is responsive and which acts to cause droplets in said jet at a droplet separation point.

12. A flow cytometer system which monitors jet conditions as described in claim 11 and further comprising a controller which is responsive to said jet condition sensor and wherein said controller acts to adjust the location at which said droplet separation point occurs.

13. A flow cytometer system which monitors jet conditions as described in claim 12 and further comprising an oscillator drive connected to said oscillator and connected to said controller.

14. A flow cytometer system which monitors jet conditions as described in claim 13 wherein said oscillator drive comprises an alternating voltage source having an amplitude and wherein said controller adjusts said amplitude.

15. A flow cytometer system which monitors jet conditions comprising:

a. a nozzle container establishing a nozzle volume and having a nozzle exit;

b. a sheath fluid port located within said nozzle volume;

c. a substance introduction port located within said nozzle volume;

d. an analysis area below said nozzle exit wherein a jet occurs;

e. a substance property sensor which senses emissions from said analysis area over a substance wavelength band; and

f. a direct jet monitor responsive to said jet and which operates simultaneously with said substance property sensor wherein said jet monitor emits a substantially separate wavelength band from said substance wavelength band.

16. A flow cytometer system which monitors jet conditions as described in claim 15 wherein said direct jet monitor comprises an optical sensor.

17. A flow cytometer system which monitors jet conditions as described in claim 16 wherein said direct jet monitor further comprises an infrared source.

18. A flow cytometer system which monitors jet conditions as described in claim 17 wherein said optical sensor comprises a charge coupled device.

19. A flow cytometer system which monitors jet conditions as described in claim 18 and further comprising:

a. an oscillator to which said sheath fluid is responsive and which acts to cause droplets in said jet at a droplet separation point;

b. an alternating voltage source having an amplitude; and

c. a controller which adjusts said amplitude in response to said direct jet monitor.

20. A flow cytometer system which monitors jet conditions as described in claim 18 and further comprising a nozzle adjuster connected to said nozzle container and responsive to said direct jet monitor which adjusts the location of said nozzle container.

21. A method of droplet flow cytometry which monitors the condition of the jet comprising the steps of:

a. establishing a nozzle volume;

b. introducing a flow of sheath fluid into said nozzle volume;

c. introducing a flow of a substance within said sheath fluid in said nozzle volume;

d. allowing said sheath fluid to exit from said nozzle volume into an analysis area to create a jet;

e. sensing a property of said substance in a substance wavelength band;

f. illuminating said jet in said analysis area in a monitor wavelength band which is substantially separate from said substance wavelength band; and

g. monitoring at least one jet condition in said analysis area in said monitor wavelength band.

22. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 21 and further comprising the step of stimulating said substance after it exits from said nozzle volume.

23. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 22 wherein said step of stimulating said substance after it exits from said nozzle volume comprises the step of causing said substance to fluoresce in a fluorescence emission band and wherein said step of sensing a property of said substance comprises the step of sensing emissions in said fluorescence emission band.

24. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 23 wherein said monitor wavelength band is substantially separate from said fluorescence emission band.

25. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 24 wherein said step of illuminating said jet in said analysis area comprises the step of subjecting said jet to infrared radiation.

26. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 25 wherein said step of monitoring at least one jet condition in said analysis area comprises the step of activating a charge coupled device.

27. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 26 wherein said step of stimulating said substance after it exits from said nozzle volume comprises the step of directing a laser at said substance.

28. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 21, 23, 25, or 27 and further comprising the step of Controlling at least one condition within said nozzle volume in response to said step of monitoring at least one jet condition in said analysis area.

29. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 28 wherein said step of controlling at least one condition within said nozzle volume in response to said step of monitoring at least one jet condition in said analysis area comprises the step of adjusting the location of said nozzle volume.

30. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 21, 23, 25, or 27 and further comprising the step of establishing an oscillation which acts to cause droplets in said jet at a droplet separation point.

31. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 30 and further comprising the step of controlling the location at which said droplet separation point occurs in response to said step of monitoring at least one jet condition in said analysis area.

32. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 31 wherein said step of establishing an oscillation is responsive to said step of controlling the location at which said droplet separation point occurs.

33. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 32 wherein said step of establishing an oscillation comprises the step of applying an alternating voltage having an amplitude to an oscillator and wherein said step of controlling the location at which said droplet separation point occurs comprises the step of varying said amplitude of said alternating voltage.

34. A method of droplet flow cytometry which monitors the condition of the jet comprising the steps of:

a. establishing a nozzle volume;

b. introducing a flow of sheath fluid into said nozzle volume;

c. introducing a flow of a substance within said sheath fluid in said nozzle volume;

d. allowing said sheath fluid to exit from said nozzle volume into an analysis area to create a jet;

e. sensing a property of said substance in a substance wavelength band; and

f. directly monitoring conditions in said jet while accomplishing said step of sensing a property of said substance in a substance wavelength band wherein said step of directly monitoring Conditions in said jet comprises the step of illuminating said jet in said analysis area in a monitor wavelength band which is substantially separate from said substance wavelength band.

35. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 34 wherein said step of directly monitoring conditions in said jet comprises the step of optically sensing said conditions from said jet without interfering with said step of sensing a property of said substance in a substance wavelength band.

36. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 35 wherein said step of directly monitoring conditions in said jet comprises the step of sensing said monitor wavelength band after it passes through a portion of said analysis area.

37. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 36 wherein said step of illuminating said jet in said analysis area in a monitor wavelength band comprises the step of subjecting said jet to infrared radiation.

38. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 36 wherein said step of sensing said monitor wavelength band after it passes through a portion of said analysis area comprises the step of activating a charge coupled device.

39. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 38 and further comprising the steps of:

a. applying an alternating voltage having an amplitude to an oscillator; and

b. adjusting said amplitude of said alternating voltage in response to said step of directly monitoring conditions in said jet.

40. A method of droplet flow cytometry which monitors the condition of the jet as described in claim 38 and further comprising the step of adjusting the location of said nozzle in response to said step of directly monitoring conditions in said jet.