Fluorescence detection system

• US 7,315,376 B2
• Filed: 01/07/2005
• Issued: 01/01/2008
• Est. Priority Date: 01/07/2005
• Status: Active Grant

• Current Assignee: GeneSTAT Molecular Diagnostics, LLC
• Original Assignee: Advanced Molecular Systems, LLC
• Inventors: Roberts, Danvern Ray, Bickmore, William D. Jr.
• Primary Examiner(s): Toatley, Jr.; Gregory J.
• Assistant Examiner(s): Skovholt; Jonathan
• Application Number:
  US11/031,526
• Publication Number:
  US 20060152727A1
• Time in Patent Office: 1,089 Days
• Field of Search: None
• US Class Current: 356/417
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates

Abstract

Apparatus and methods for exciting and detecting fluorescence in samples are disclosed. In one embodiment, a sample holder for holding a plurality of samples is provided together with an optical manifold having an excitation source, a photo receiver, or both, for each of the plurality of samples. In another embodiment, the optical manifold contains only the excitation source or a photo receiver, and the other is associated with the sample holder. This system permits for rapid excitation and measurement of fluorescence without the use of moving parts and without any opto-mechanical or electronic disturbance. It exhibits an exceptional signal to noise ratio, which permits it to differentiate between very low level differences in fluorescence.

Claims

1. A system for detecting fluorescence, comprising:

a sample holder for holding a plurality of samples, at least some of which may contain a fluorescent material;

a plurality of excitation sources, one for each of the plurality of samples, capable of exciting the fluorescent material;

a plurality of photo receivers, one for each of the plurality of samples, capable of detecting fluorescent emissions from the fluorescent material when it is excited;

an emission bandpass filter for each of the plurality of photo receivers, each said emission bandpass filter being positioned perpendicular to the axis of reception of the photo receiver with which it is associated, with no further optical elements between the associated photo receiver and the associated sample;

an optical manifold for holding each excitation source and each photo receiver in association with the sample holder, so that operation of each excitation source will excite any of said fluorescent material within the corresponding sample and be detected by the corresponding photo receiver.

2. The system of claim 1, wherein each excitation source and corresponding photo receiver are offset off axis so as to fit in close proximity to one another while defining a ray trace from excitation source to the sample and back to the corresponding photo receiver.

3. The system of claim 1, wherein each excitation source and corresponding photo receiver are offset 7 degrees off axis.

4. The system of claim 1, further comprising an amplifier associated with each of the photo-receivers.

5. The system of claim 4, wherein each of the amplifiers are provided with an adjustable gain control.

6. The system of claim 1, wherein the illumination of the excitation source is proportional to the voltage applied, and further comprising a variable voltage control for each of the plurality of excitation sources.

7. The system of claim 1, wherein each of the photo receivers comprise an avalanche receiver.

8. The system of claim 1, further comprising a controller for passing data sequentially from the plurality of photo receivers to a recording device.

9. The system of claim 1, wherein the sample holder is configured to receive a plurality of sample vials.

10. The system of claim 1, wherein the sample holder contains a plurality of dimples for holding sample.

11. The system of claim 1, wherein the photo receivers comprise a high impedance cadmium sulfide photo resistor, and further comprising a high impedance dropping resistor for development of an electrical signal.

12. The system of claim 11, further comprising a filter capacitor for dampening electronic radio frequency interference.

29. A system for detecting fluorescence as defined in claim 1, further comprising an excitation bandpass filter for each of the plurality of excitation sources, each said excitation bandpass filter being positioned perpendicular to the axis of emission of the excitation source with which it is associated, with no further optical elements between the associated excitation source and the associated sample.

30. A system for detecting fluorescence as defined in claim 1, wherein each excitation source has a corresponding photo receiver, and the axis of corresponding excitation source and its associated excitation bandpass filter is offset from the axis of the corresponding photo receiver and its associated emission bandpass filter by 180 degrees.

31. A system for detecting fluorescence as defined in claim 29, wherein each excitation source has a corresponding photo receiver, and the axis of corresponding excitation source and its associated excitation bandpass filter is offset from the axis of the corresponding photo receiver and its associated emission bandpass filter by 180 degrees.

13. A system for detecting fluorescence, comprising:

a sample holder for holding a plurality of samples;

a plurality of excitation sources, one for each of the plurality of samples;

a plurality of photo receivers, one for each of the plurality of samples;

an emission bandpass filter for each of the plurality of photo receivers, each said emission bandpass filter being positioned perpendicular to the axis of reception of the photo receiver with which it is associated, with no further optical elements between the associated photo receiver and the associated sample;

an optical manifold;

the optical manifold being configured for holding either the plurality of excitation sources or the plurality of photo receivers in association with the sample holder, and the sample holder being configured so as to hold the other of the plurality of excitation sources or the plurality of photo receivers, so that operation of each excitation source will excite any of said fluorescent material within the corresponding sample and be detected by the corresponding photo receiver.

14. The system of claim 13, wherein the optical manifold holds the plurality of excitation sources and the sample holder holds the plurality of photo receivers.

15. The system of claim 13, wherein the optical manifold holds the plurality of photo receivers and the sample holder holds the plurality excitation sources.

16. The system of claim 13, further comprising an amplifier associated with each of the photo-receivers.

17. The system of claim 16, wherein each of the amplifiers are provided with an adjustable gain control.

18. The system of claim 13, wherein the illumination of the excitation source is proportional to the voltage applied, and further comprising a variable voltage control for each of the plurality of excitation sources.

19. The system of claim 13, wherein each of the photo receivers comprise an avalanche receiver.

20. The system of claim 13, further comprising a controller for passing data sequentially from the plurality of photo receivers to a recording device.

21. The system of claim 13, wherein the sample holder is configured to receive a plurality of sample vials.

22. The system of claim 13, wherein the sample holder contains a plurality of dimples for holding sample.

23. The system of claim 13, wherein the photo receivers comprise a high impedance cadmium sulfide photo resistor, and further comprising a high impedance dropping resistor for development of an electrical signal.

24. The system of claim 13, further comprising a filter capacitor for dampening electronic radio frequency interference.

32. A system for detecting fluorescence as defined in claim 13, further comprising an excitation bandpass filter for each of the plurality of excitation sources, each said excitation bandpass filter being positioned perpendicular to the axis of emission of the excitation source with which it is associated, with no further optical elements between the associated excitation source and the associated sample.

25. A system for detecting fluorescence, comprising:

a sample holder for holding a plurality of samples;

a plurality of excitation sources, one for each of the plurality of samples;

a plurality of photo receivers, one for each of the plurality of samples;

an emission bandpass filter for each of the plurality of photo receivers, each said emission bandpass filter being positioned perpendicular to the axis of reception of the photo receiver with which it is associated, with no further optical elements between the associated photo receiver and the associated sample;

the sample holder being configured for holding said plurality of excitation sources and plurality of photo receivers so that operation of each excitation source will excite any of said fluorescent material within the corresponding sample and be detected by the corresponding photo receiver.

26. The system of claim 25, wherein the photo receivers comprise a high impedance cadmium sulfide photo resistor, and further comprising a high impedance dropping resistor for development of an electrical signal.

27. The system of claim 25, further comprising a filter capacitor for dampening electronic radio frequency interference.

33. A system for detecting fluorescence as defined in claim 25, further comprising an excitation bandpass filter for each of the plurality of excitation sources, each said excitation bandpass filter being positioned perpendicular to the axis of emission of the excitation source with which it is associated, with no further optical elements between the associated excitation source and the associated sample.

28. A method for detecting fluorescence in a plurality of samples, comprising:

providing a plurality of samples including fluorescent material;

providing an excitation source for each of the plurality of samples for exciting said fluorescent material;

providing a photo receiver for each of the plurality of samples for detecting fluorescent emissions from said fluorescent material when it is excited;

providing an emission bandpass filter for each of the plurality of photo receivers, each said emission bandpass filter being positioned perpendicular to the axis of reception of the photo receiver with which it is associated, with no further optical elements between the associated photo receiver and the associated sample;

activating each excitation source; and

measuring the output of each photo receiver.

34. A method for detecting fluorescence as defined in claim 28, further comprising the step of providing an excitation bandpass filter for each of the plurality of excitation sources, each said excitation bandpass filter being positioned perpendicular to the axis of emission of the excitation source with which it is associated, with no further optical elements between the associated excitation source and the associated sample.