Multi parameter scanner

• US 6,043,506 A
• Filed: 12/30/1997
• Issued: 03/28/2000
• Est. Priority Date: 08/13/1997
• Status: Expired due to Term

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Bio-Rad Laboratories Incorporated
• Inventors: Heffelfinger, David M., Salgado, Renato, Batterson, Rebecca Ann
• Primary Examiner(s): Westin, Edward P.
• Assistant Examiner(s): HANIG, RICHARD E
• Application Number:
  US09/001,254
• Time in Patent Office: 819 Days
• Field of Search: 250/584, 250/585, 250/586, 250/458.1, 250/484.4, 356/417
• US Class Current: 250/584
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 2021/6421 Measuring at two or more wavelengths
• CPC Class Codes: G01N 2021/6471 Special filters, filter wheel
• CPC Class Codes: G01N 2021/6484 Optical fibres
• CPC Class Codes: G01N 2201/024 Modular construction
• CPC Class Codes: G03B 42/00 Obtaining records using waves other than optical waves; Visualisation of such records by using optical means

Abstract

An apparatus capable of measuring quantities of biological or other types of samples that have been labeled using any of a variety of techniques including fluorescence, radioisotopes, enzyme activated light emitting chemicals, and enzyme activated fluorescent materials is provided. The apparatus allows for either simultaneous or sequential acquisition of signals from multiple sample types. The apparatus is not restricted to a particular source or wavelength of excitation or readout light, nor is the apparatus restricted to a particular emission wavelength. The provided scanner includes a source module that preferably contains an internal laser emitting two different wavelengths of approximately the same intensity. An optional external light source may be coupled to the source module, thus adding further flexibility through the addition of other wavelengths (e.g., V, visible, mid-IR, and IR). The scanner also includes a detection module. Within the detection module are two detectors, thus allowing the simultaneous detection of multiple wavelengths. A bifurcated optical cable is used to transfer the excitation and/or readout light from the source module to the sample and subsequently transfer the emitted and/or scattered light from the sample to the detection module. The scanning stage of the scanner is designed to accommodate a variety of samples, ranging from phosphor screens, gels, and fluorescent samples to microtiter plates. An internal microprocessor is used to control the various aspects of the scanner, preferably including translation stage control, source filters, and detection filters. The internal microprocessor may be coupled to an external computer. The external computer may be used to change the programming of the microprocessor, provide a user interface to the microprocessor, process and store test results, and display sample images.

Claims

1. A scanner system, comprising:

a sample illumination source, wherein said illumination source is comprised of a laser, wherein said laser simultaneously emits radiation of a first wavelength and a second wavelength;

a scan head for scanning an area of a sample held within a sample support, wherein said scanning is along a first direction and a second direction, wherein said scan head directs said radiation from said source to a portion of said sample;

a detection system for monitoring radiation emitted by said sample, wherein said detection system comprises a first detector monitoring radiation of a third wavelength and a second detector monitoring radiation of a fourth wavelength, wherein said first and second detectors simultaneously monitor radiation emitted by said sample, and wherein each detector outputs a signal corresponding to the intensity of said radiation; and

a wavelength selection system coupled to said detection system, wherein said wavelength selection system determines said third and fourth wavelengths of radiation monitored by said first and second detectors.

2. The scanner system of claim 1, further comprising:

a first translation stage for moving said scan head in said first direction;

a second translation stage for moving said scan head in said second direction; and

a processor coupled to said first and second translation stages, said processor controlling movement of said scan head.

3. The scanner system of claim 2, further comprising a third translation stage coupled to said scan head, said third translation stage controlling motion of said scan head in a third direction, wherein said third direction is perpendicular to a plane formed by said first and second directions, and wherein said processor controls movement of said third translation stage.

4. The scanner system of claim 1, wherein said first and second detectors are photomultiplier tube detectors.

5. The scanner system of claim 4, further comprising a processor coupled to said first and second detectors, wherein said processor controls a first gain associated with said first detector and a second gain associated with said second detector.

6. The scanner system of claim 1, wherein a first intensity of said laser radiation of said first wavelength is approximately equal to a second intensity of said laser radiation of said second wavelength.

7. The scanner system of claim 1, wherein said wavelength selection system is further comprised of at least a beam splitter and a bandpass filter.

8. The scanner system of claim 1, wherein said wavelength selection system is further comprised of at least two beam splitters and at least two bandpass filters.

9. The scanner system of claim 1, further comprising a bifurcated bundle of optical fibers having distal and proximal ends, said optical fibers comprising at least one emitter fiber and at least one collecting fiber, said distal ends of said at least one emitter fiber and said at least one collecting fiber coupled to said scan head, wherein said radiation from said illumination source is focussed by a first focussing optic into said proximal end of said at least one emitter fiber, said radiation passing through said at least one emitter fiber and being focussed by a second focussing optic onto said portion of said sample, and wherein said radiation emitted by said portion of said sample is collected by a collection optic and focussed onto said distal end of said at least one collecting fiber, and wherein said proximal end of said at least one collecting fiber is positioned to pass said collected radiation to said detection system through said wavelength selection system.

10. The scanner system of claim 1, further comprising a filter wheel comprised of at least a plurality of optical bandpass filters, wherein radiation from said illumination source passes through said filter wheel.

11. The scanner system of claim 10, wherein said filter wheel is further comprised of at least one opaque optical shutter element.

12. The scanner system of claim 1, further comprising a processor coupled to said detection system, wherein said processor creates an image of said scan area of said sample from said detection system output signals.

13. The scanner system of claim 12, further comprising a monitor coupled to said processor, wherein said image is displayed on said monitor.

14. The scanner system of claim 1, wherein said sample illumination source further comprises an external source port allowing an external source to be coupled to said scanner system, wherein radiation from said external source passes through said external source port and said scan head to said portion of said sample.

15. The scanner system of claim 14, further comprising said external source coupled to said external source port, wherein said external source is selected from the group consisting of lasers and broadband sources.

16. The scanner system of claim 14, further comprising:

a radiation collimating system, wherein radiation from said external source passes through said collimating system; and

a translation stage system coupled to said collimating system, said translation stage system allowing said external source radiation to be optimally coupled to said scanner system.

17. The scanner system of claim 16, further comprising a processor coupled to said translation stage system, said processor controlling movement of said translation stage system and optimization of said coupling of said external source radiation to said scanner system.

18. The scanner system of claim 17, wherein said coupling optimization is determined by peaking said output signal from said detection system during movement of said translation stage system.

19. The scanner system of claim 17, wherein said coupling optimization is determined by peaking a signal from an alignment detector during movement of said translation stage system.

20. The scanner system of claim 15, wherein radiation from said external source and said laser simultaneously illuminate said sample.

21. The scanner system of claim 1, wherein said radiation emitted by said sample is scattered radiation originating with said illumination source.

22. The scanner system of claim 1, wherein said radiation emitted by said sample is due to fluorescence, wherein said fluorescence is of a wavelength distinguishable from said first and second wavelengths of said illumination source.

23. The scanner system of claim 1, wherein said sample is selected from the group consisting of fluorescent samples, storage phosphor screens, samples containing DNA stains, samples containing chemiluminescent probes, and samples containing protein dyes.

24. The scanner system of claim 1, wherein said sample is selected from a group of sample types consisting of gels, membranes, TLC plates, sequencing plates, DNA hybridization arrays, and PCR chips.

25. The scanner system of claim 1, wherein said first illumination source wavelength is suitable for use with storage phosphor screens based on BaFBr:

Eu and said second illumination source wavelength is suitable for use with storage phosphor screens based on SrS;

Ce and SrS;

Sm.

26. The scanner system of claim 2, further comprising a first optical encoder coupled to said first translation stage and a second optical encoder coupled to said second translation stage, wherein said first and second optical encoders provide absolute position information for said scan head.

27. The scanner system of claim 1, wherein said illumination source is modulated and said detection system is modulated.

28. The scanner system of claim 27, further comprising a processor coupled to said modulated detection system, wherein said illumination source excites fluorescence in said sample, and wherein said processor determines fluorescence lifetimes from said output signals of said modulated detection system.

29. A storage phosphor screen scanner, comprising:

a laser source simultaneously emitting radiation at a first wavelength and a second wavelength, wherein said first wavelength is compatible with BaFBr;

Eu storage phosphor screens and said second wavelength is compatible with SrS;

Ce and SrS;

Sm storage phosphor screens;

a screen scanning head coupled to said laser source by at least one excitation optical fiber, wherein said scanning head is coupled to an x-y scanning system;

a detection system comprised of a first detector monitoring radiation of a third wavelength and a second detector monitoring radiation of a fourth wavelength and a beam splitter transmitting radiation of said third wavelength to said first detector and reflecting radiation of said fourth wavelength to said second detector, wherein said first and second detectors simultaneously monitor radiation, and wherein said detection system is coupled to said scanning head by at least one collection optical fiber; and

a processor coupled to said x-y scanning system, said processor controlling an area of a sample scanned by said scanning head.

30. The storage phosphor screen scanner of claim 29, wherein said processor controls a resolution and a scan speed associated with said x-y scanning system.

31. The storage phosphor screen scanner of claim 29, wherein said first wavelength is approximately 532 nanometers and said second wavelength is approximately 1064 nanometers.

32. The storage phosphor screen scanner of claim 29, further comprising an external source coupled to said scanning head through said at least one excitation optical fiber, wherein said external source emits radiation of a fifth wavelength distinguishable from said first and second wavelengths.

33. The storage phosphor screen scanner of claim 29, further comprising a sample storage phosphor screen, said sample storage phosphor screen exposed in a substantially light tight exposure box prior to insertion into said storage phosphor screen scanner.

34. The storage phosphor screen scanner of claim 29, further comprising a wavelength selection system interposed between said laser and said at least one excitation optical fiber.

35. The storage phosphor screen scanner of claim 29, further comprising a wavelength selection system interposed between said at least one collection optical fiber and said first and second detectors.

36. The storage phosphor screen scanner of claim 29, wherein said processor is coupled to said detection system, and wherein said processor creates an image of a sample within said storage phosphor screen scanner.

37. A scanner system, comprising:

a sample illumination source, said illumination source comprising;

an internal source emitting radiation of at least a first wavelength; and

an external source emitting radiation of at least a second wavelength;

a radiation collimating system, wherein radiation from said external source passes through said collimating system;

a translation stare system coupled to said collimating system, said translation stage system allowing said external source radiation to be optimally coupled to said scanner system;

a processor coupled to said translation stage system, said processor controlling movement of said translation stage system and optimization of said coupling of said external source radiation to said scanner system;

a scan head for scanning an area of a sample held within a sample support, wherein said scanning is along a first direction and a second direction, wherein said scan head simultaneously directs said radiation of said at least first and second wavelengths from said sample illumination source to a portion of said sample;

a detection system for monitoring radiation emitted by said sample, wherein said detection system outputs a signal corresponding to the intensity of said monitored radiation; and

a wavelength selection system coupled to said detection system, wherein said wavelength selection system controls the wavelength of radiation monitored by said detection system.

38. The scanner system of claim 37, further comprising:

a first translation stage for moving said scan head in said first direction;

a second translation stage for moving said scan head in said second direction; and

a processor coupled to said first and second translation stages, said processor controlling movement of said scan head.

39. The scanner system of claim 38, further comprising a third translation stage coupled to said scan head, said third translation stage controlling motion of said scan head in a third direction, wherein said third direction is perpendicular to a plane formed by said first and second directions, and wherein said processor controls movement of said third translation stage.

40. The scanner system of claim 37, wherein said detection system is further comprised of a first detector and a second detector, and wherein said scanner system further comprises a beam splitter, wherein said beam splitter allows said first detector and said second detector to simultaneously monitor radiation emitted by said sample, wherein said first detector monitors radiation of a third wavelength and said second detector monitors radiation of a fourth wavelength.

41. The scanner system of claim 40, wherein said first and second detectors are photomultiplier tube detectors.

42. The scanner system of claim 37, wherein a first intensity of said internal source emissions is approximately equal to a second intensity of said external source emissions.

43. The scanner system of claim 37, further comprising a bifurcated bundle of optical fibers having distal and proximal ends, said optical fibers comprising at least one emitter fiber and at least one collecting fiber, said distal ends of said at least one emitter fiber and said at least one collecting fiber coupled to said scan head, wherein said radiation from said sample illumination source is focussed by a first focussing optic into said proximal end of said at least one emitter fiber, said radiation passing through said at least one emitter fiber and being focussed by a second focussing optic onto said portion of said sample, and wherein said radiation emitted by said portion of said sample is collected by a collection optic and focussed onto said distal end of said at least one collecting fiber, and wherein said proximal end of said at least one collecting fiber is positioned to pass said collected radiation to said detection system through said wavelength selection system.

44. The scanner system of claim 37, further comprising a processor coupled to said detection system, wherein said processor creates an image of said scan area of said sample from said detection system output signals.

45. The scanner system of claim 38, further comprising a first optical encoder coupled to said first translation stage and a second optical encoder coupled to said second translation stage, wherein said first and second optical encoders provide position information for said scan head.