Optical biosensor system

• US 5,313,264 A
• Filed: 05/10/1991
• Issued: 05/17/1994
• Est. Priority Date: 11/10/1988
• Status: Expired due to Term

• Current Assignee: GE Healthcare Bio-Sciences AB (General Electric Company)
• Original Assignee: Pharmacia Biosensor AB
• Inventors: Sjodin, Hakan, Sjolander, Stefan, Ulf, Jonsson, St.ang.hlberg, Ralph, Ivarsson, Bengt
• Primary Examiner(s): Evans, F. L.
• Application Number:
  US07/681,533
• Time in Patent Office: 1,103 Days
• Field of Search: 356/73, 356/317, 356/318, 356/338, 356/343, 356/417, 356/446, 356/246, 356/445, 250/458.1, 250/459.1, 250/461.1, 250/461.2
• US Class Current: 356/73
• CPC Class Codes: G01N 2021/0346 Capillary cells; Microcells
• CPC Class Codes: G01N 2021/212 Arrangement with total internal reflection
• CPC Class Codes: G01N 2021/5957 using an image detector type detector, e.g. CCD
• CPC Class Codes: G01N 21/0332 with temperature control control of temperature G05D23/00; cryostats F17C3/08
• CPC Class Codes: G01N 21/05 Flow-through cuvettes G01N21/09 takes precedence; handling fluid samples G01N1/10
• CPC Class Codes: G01N 21/552 Attenuated total reflection
• CPC Class Codes: G01N 21/553 and using surface plasmons fluorescence excitation G01N21/648; enhanced Raman G01N21/658
• CPC Class Codes: G01N 21/648 using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• CPC Class Codes: G02B 6/34 utilising prism or grating G02B6/293 takes precedence

Abstract

An optical biosensor system using internal reflection versus angle of incidence determination for the detection of biomolecules, the system comprising a sensor unit (10) with at least two sensing surfaces (39A-D), a source of light (1), and lens means (2) for forming a convergent beam of light which is focused in wedge-shape fashion to form a streak of light (5) extending transversely over all the sensing surfaces; a photodetector device (7) in the form of a two-dimensional matrix of individual photodetector; optical imaging instrumentation in the form of an anamorphic lens system (6) for the purpose of imaging rays of reflected light from the sensing surfaces on each its own column of photodetectors, so that for each sensing surface there is a corresponding set of columns of photodetectors; and an evaluation unit (8) for determining the minimum reflectance or the resonance angle at each of the sensing surfaces. The invention also relates to a method for calibrating the biosensor system, a method for correcting for baseline drift as well as a method for temperature regulation of thermostat means in the biosensor system.

Claims

1. An optical biosensor system using an internal reflection versus angle of incidence determination for detecting a specific biomolecule, comprising:

a sensor unit including a transparent plate, at least a portion of one of the faces of said plate having a dielectric film adhered thereto, or being primarily coated with a metal film to which has been adhered a dielectric film, said dielectric film having an affinity for the specific biomolecule at the time the measurement is carried out;

a light source, including first lens means, for directing a beam of light at an interface of the transparent plate and said metal film or said dielectric film directly adhered thereto to produce an evanescent wave at a sensing surface of said dielectric film when a sample solution contacts said dielectric film on the transparent plate;

optical imaging instrumentation including second lens means by which one of light reflected from said sensing surface of the transparent plate, light originating from the sample through evanescent wave stimulated fluorescence or phosphorescence, and light originating from scattering at said sensing surface, is imaged on a photodetector device; and

an evaluation unit for determining at least one of an angle of minimum reflectance, a state of polarization, and a collected light intensity;

wherein at least one sensing surface is arranged on the sensor unit;

wherein said at least one sensing surface, in order to form an elongated flow cell, is pressed over an upwardly open portion of at least one channel for the sample solution which is to be analyzed and affects at least one of the angle of minimum reflectance, the polarization state for minimum reflectance, and the collected light intensity, at which the evanescent wave interacts with the sample;

wherein said first lens means forms a convergent beam of light focused in a wedge-shape fashion to form a streak of light extending transversely over all of said at least one sensing surface;

wherein the photodetector device, at least in the case of more than one sensing surface, is a two-dimensional matrix of individual photodetectors;

wherein said second lens means is a lens system for imaging rays of one of said reflected light from said at least one sensing surface, said light emitted from the sample, and said light originating from scattering at the sensing surface onto a corresponding column of photodetectors, so that said at least one sensing surface has at least one corresponding column of photodetectors;

wherein said evaluation unit determines at least one of the angle of minimum reflectance, the state of polarization, and the collected light intensity, for said at least one sensing surface.

2. The optical biosensor system of claim 1, further comprising:

a support in which the open portion of the at least one channel is arranged in a fixed position;

a housing in which said light source, said optical imaging instrumentation, the photodetector device, and a coupling prism are arranged in fixed positions;

an aperture provided in said housing below said coupling prism and covered by an opto-interface plate for coupling incident and reflected light and/or scattered or emitted light from the sample, respectively, to said sensor unit and said optical imaging instrumentation; and

holder means by which said sensor unit is detachably attached below said aperture,

said housing being placed on top of said sensor unit for pressing the opto-interface plate against an opposite face of said sensor unit.

3. The optical biosensor system of claim 2, wherein said housing is hinged to said support by means of a system of articulated arms extending from each lateral face of said housing and said support, respectively.

4. The optical biosensor system of claim 2, further comprising thermostat means provided in said housing and at the at least one channel, for thermostatically controlling said at least one sensing surface and the at least one channel, respectively.

5. The optical biosensor system of claim 1, wherein said at least one sensing surface includes a plurality of sensing surfaces, arranged in a side-by-side relationship on said sensor unit.

6. The optical biosensor system of claim 1, wherein the transparent plate of said sensor unit is of glass.

7. The optical biosensor system of claim 1, wherein the dielectric film comprises a dextran layer.

8. The optical biosensor system of claim 1 wherein the at least one channel includes a plurality of channels arranged in a parallel side-by-side relationship, wherein each of the plurality of channels has a corresponding upwardly open portion, wherein the corresponding upwardly open portions are all covered by said sensor unit when a measurement is performed, and wherein in said measuring stage, wherein each of said at least one sensing surfaces is situated above its corresponding upwardly open portion.

9. The optical biosensor system of claim 8, wherein each of the corresponding upwardly open portions are slits extending through a first layer of elastomer material or silicon.

10. The optical biosensor system of claim 9, wherein the first layer is made of a rubber material.

11. The optical biosensor system of claim 10, wherein said plurality of channels are formed in a liquid handling block unit of a solid material selected from the group consisting of plastics, metals and ceramics.

12. The optical biosensor system of claim 8 wherein means are provided which permit said plurality of channels to be coupled in series with each other.

13. The optical biosensor system of claim 9, wherein said plurality of channels are formed in a liquid handling block unit of a solid material selected from the group consisting of plastic, metal, and ceramics, and wherein said liquid handling block unit comprises:

a planar support plate having a second elastomer layer on its upper face;

a base plate positioned on top of the second elastomer layer including,

a third elastomer layer positioned on an underside of said base plate,

a first number of riser ducts which correspond to a number of the corresponding upwardly open channel portions, extend through said base plate, and connect with one end of each of the corresponding upwardly open channel portions, and

a second number of riser ducts which correspond to the number of corresponding upwardly open channel portions and connect with an opposite end of each of the corresponding upwardly open channel portions;

a pattern of channels formed in the third elastomer layer including,

at least one primary volume for a sample liquid, said primary volume having a first exactly determined volume,

at least one inlet channel for said sample liquid,

at least one inlet channel for eluent liquid, and

an outlet channel for said sample and eluent liquids; and

a plurality of individually controllable valve means in said planar support plate for directing the liquid flow in said plurality of channels.

14. The optical biosensor system of claim 13, further comprising a secondary volume for said sample liquid, said secondary volume having a second exactly determined volume which differs from said first exactly determined volume.

15. The optical biosensor system of claim 14, further comprising a second plurality of channels so that said second plurality of channels are filled while said sample liquid of said plurality of channels is analyzed.

17. The optical biosensor system of claim 13, wherein each of said plurality of individually controllable valve means comprise solenoids actuating respective wires, each wire being of superelastic material and extending through the through opening in said support plate to act upon the second elastomer layer.

18. The optical biosensor system of claim 8, wherein the corresponding upwardly open portions have a length of about 800 μ

m, a width of about 300 μ

m, and a height of about 30 μ

m.

19. The optical biosensor system of claim 2, further comprising:

a carrier plate for said sensor unit, wherein an opening in said carrier plate accommodates said sensor unit and wherein said sensor unit rests on flange means at a lower end of said opening.

20. The optical biosensor system of claim 1, wherein said second lens means is an anamorphic second lens system for

imaging reflected rays of light from surface elements in a longitudinal direction of the streak of light, in the form of real image elements corresponding to rows and columns of photodetectors where each column corresponds to part of said at least one sensing surface in a direction of the streak of light, and

imaging reflected rays of light from surface elements transverse to the longitudinal direction of the streak of light, onto pixels situated along the columns of photodetectors, such that mutually parallel incident rays lying in a plane of incidence and aligned transversely to the longitudinal direction of the streak of light are focused toward a single photodetector in a column, so that each detector in a column corresponds to a one particular angle of incidence.

21. The optical biosensor system of claim 20, wherein said internal reflection versus angle of incidence determination is based upon surface plasmon resonance.

22. The optical biosensor system of claim 21, wherein a convergent beam of light strikes a reverse side of the metal film with angles of incidence of between 62 and 78 degrees.

23. The optical biosensor system of claim 1, wherein said second lens means is arranged for collecting one of rays of light emitted from a sample and rays of light scattered from surface elements in a longitudinal direction of the streak of light, onto columns of photodetectors where each column corresponds to part of said at least one sensing surface in the direction of the streak of light, and for

collecting rays of light from surface elements transverse to the longitudinal direction of the streak of light, onto pixels situated along the columns of photodetectors, such that mutually parallel incident rays lying in a plane of incidence and aligned transverse to the longitudinal direction of the streak of light are focused toward a single photodetector in a column, so that each photodetector in a column corresponds to one particular angle of incidence.

24. The optical biosensor system of claim 1, wherein said at least one sensing surface includes a plurality of surfaces, a width of the streak of light irradiating said plurality of sensing surface is substantially equal to an extent of said plurality of sensing surfaces transverse to the longitudinal direction of the streak of light to produce an integrated mean value of a mass of biomolecules binding to the dielectric film transverse to the longitudinal direction of the streak of light, thereby permitting a reproducible quantitative analysis or alternatively a reproducible analysis of the relative amount of the particular biomolecule.

25. The optical biosensor system of claim 24, wherein an extent of said plurality of sensing surfaces in the longitudinal direction of the streak of light are imaged in reduced size on a single photodetector, to produce an integrated mean value of the mass of biomolecules binding to the dielectric film in the longitudinal direction of the streak of light.

26. The optical biosensor system of claim 20, wherein said second lens means reduces an image element in S planes with a linear degree of magnification m=0.56 when the extent is of the order of magnitude of about 300 μ

m and the photodetector has dimensions of about 90×

90 μ

m.

27. The optical biosensor system of claim 1, wherein said first lens means of said light source includes a lens member for varying the width of the streak of light irradiating each of said at least one sensing surfaces.

28. The optical biosensor system of claim 2, wherein said coupling prism is a truncated straight hemicylinder placed over said aperture in a bottom wall of the housing.

29. The optical biosensor system of claim 22, wherein said light source in combination with a filter emits substantially monochromatic and incoherent light.

30. The optical biosensor system of claim 22, wherein said light source emits substantially monochromatic and coherent light.

31. The optical biosensor system of claim 2, further comprising positioning means for determining relative positions of said housing, said sensor unit and said support.

32. The optical biosensor system of claim 2 wherein said opto-interface plate includes,

a second transparent plate of glass including longitudinally extending parallel ridges on one face aligned with longitudinally extending parallel ridges on an opposite face of said second transparent plate, said ridges being made of an elastic transparent material, spaced apart with distances between corresponding to distances between each of said at least one sensing surfaces with a length at least sufficient to couple an entire cross section of the convergent beam of light to said sensor unit and pressed against said transparent plate of said sensor unit directly opposite each of said at least one sensing surfaces.

33. The optical biosensor system of claim 32, wherein each of said longitudinally extending parallel ridges has a number of longitudinally extending stepped portions on each side to form a structure having, in cross section, a configuration of a flight of stairs, the uppermost steps thereof being pressed resiliently against said sensor unit and said coupling prism without formation of enclosed air pockets.

34. The optical biosensor system of claim 23, wherein said evaluation unit is arranged such that when the mass of bound specific biomolecules is calculated, the calculation includes light emitted from samples or scattered from said at least one sensing surface from only a partial band of the width of said at least one sensing surface and positioned centrally therein, this positioning being effectuated by evaluation of electric signals obtained from the photodetectors in a column which corresponds to a corresponding partial band on each of said at least one sensing surface.

35. The optical biosensor system of claim 25, wherein said evaluation unit is arranged such that when the mass of bound specific biomolecules is calculated, the calculation includes reflected light from only a narrow band.

36. The optical biosensor system of claim 35, wherein said evaluation unit is arranged for calculating parameters for a surface plasmon resonance curve for a determination including a reflectance minimum and a resonance angle and a displacement thereof for each flow cell, by curve fitting response values obtained from column detector members corresponding to the resonance angle.

37. The optical biosensor system of claim 2, wherein said coupling prism is a plane-sided prism placed over the aperture in a bottom wall of said housing, wherein refractive properties of said plane-sided prism determine a layout of said first lens means in order to form a convergent beam of light focused in a wedge-shaped fashion into the streak of light extending transversely over all sensitized areas.

38. The optical biosensor system of claim 37, wherein the upwardly open channel portion of at least one channel is a wall-jet cell, from which a jet flow is directed against a centre of a circular sensing surface.

39. A method of calibrating the optical biosensor system of claim 1, comprising the steps of:

directing a beam of light from said light source to form a streak of light extending transversely over all of said at least one sensing surface;

pumping a plurality of calibration solutions without affinity to said at least one sensing surface, each with a known refractive index one after the other, through a flow cell,

storing electric signals from the detectors of the photodetector rows corresponding to said at least one sensing surface in a memory, for each of the plurality of calibration solutions, so that a known relationship is obtained between a resonance angle and a refractive index of the dielectric film, and

calibrating the response from each of the photodetector rows.

40. The method of claim 39, further comprising the steps of:

passing the sample solution over a separate sensing surface that has no affinity for any biomolecule and is situated on said metal film;

storing the electric signals obtained from the photodector rows corresponding to the separate, non-affinity sensing surface in another location of the memory;

and correcting the stored signals obtained when the sample solution is passed along said at least one sensing surface, to determine a difference between the refractive indices due to the presence of the particular biomolecule for said at least one sensing surface.

41. A method of correcting for baseline drift in surface plasmon resonance (SPR) analysis procedures using the optical biosensor system of claim 1, comprising the steps of:

establishing a predetermined temperature set point for the sample solution;

creating an electric signal representing an actual temperature value of the sample solution, by utilizing electric signals which represent the resonance angle as measured in a flow cell, a sensing surface of which has no affinity for the specific molecule and through which a liquid is fed which is conveyed through a liquid flow handling unit;

correcting the signals from the sample solution with respect to a temperature disturbance caused by not positioning a thermostat in the flow cell; and

controlling said thermostat by measuring difference between the corrected signal and the actual value in order to adjust the temperature set point.

42. A method of measuring the actual feedback value of the temperature at the flow channels for regulation of a thermostat system by surface plasmon resonance (SPR) analysis procedures using the optical biosensor system of claim 1 comprising the steps of:

creating an electrical signal representing an actual temperature value of the sample solution, by utilizing electric signals which represent the resonance angle as measured at a flow cell, a sensing surface of which is in contact with at least one sealing area adjacent or between the flow channels, wherein said sealing area is coated with a dielectricum of suitable material and refractive index properties in order to obtain a surface plasmon resonance (SPR) response from said sealing area, said dielectricum having a known temperature dependence of its refractive index, light beams reflected at areas of the metal film opposite to areas of the film pressed in close optical contact with the liquid sealing areas of said dielectricum being imaged by the second lens means, being anamorphic onto corresponding columns of detector elements; and

controlling said thermostat system by measuring a difference between the corrected signal and the actual value in order to adjust the temperature toward the temperature set point.

43. The optical biosensor system of claim 1, wherein said optical imaging instrumentation further including optical filter means.

44. The optical biosensor system of claim 10, wherein the rubber material is silicon rubber.

45. The optical biosensor system of claim 29, wherein said light source includes a light emitting diode and a interference filter.

46. The optical biosensor system of claim 30, wherein said light source includes one of a semiconductor diode laser, a dye laser, and a gas laser.

47. The optical biosensor system of claim 36, wherein the response values are curve fitted using a reflectance minimum of the resonance angle.

48. The optical biosensor system of claim 1, wherein said at least one sensing surface includes a plurality of surfaces, a width of the streak of light irradiating said plurality of sensing surfaces is substantially equal to an extent of said plurality of sensing surfaces transverse to the longitudinal direction of the streak of light to produce an integrated mean value of a mass of biomolecules binding to the dielectric film transverse to the longitudinal direction of the streak of light, thereby permitting a reproducible quantitative analysis or alternatively a reproducible analysis of the relative amount of the particular biomolecule.

49. The optical biosensor system of claim 43, wherein an extent of each of said at least one sensing surfaces in the longitudinal direction of the streak of light are imaged in reduced size on a single photodetector, to produce an integrated mean value of the mass of biomolecules binding to the dielectric film in the longitudinal direction of the streak of light.

50. The optical biosensor system of claim 1, wherein said second lens means is an anamorphic lens system for imaging rays of reflected light from said at least one sensing surface, and said evanescent wave is produced by surface plasmon resonance.

51. The optical biosensor system of claim 1, wherein said second lens means is a lens system for imaging one of rays of light emitted from the sample and rays scattered at the sensing surface, said emission and said scattering of said light rays being generated by said evanescent wave.

52. The optical biosensor system of claim 35, wherein said narrow band is preferably about 50% of the width of each of said plurality of sensing surfaces, said narrow band extending substantially along an entire length of each of said plurality of sensing surfaces and positioned centrally therein, this positioning being effectuated by evaluation of electric signals obtained from the photodetectors in a column which corresponds to a corresponding narrow band on each of said plurality of sensing surfaces.

53. The method of claim 39, wherein said plurality of calibration solutions are salt solutions and/or suitable organic solutions.

16. The optical biosensor system of 13, wherein pneumatic valves are formed in through openings in said planar support plate, by the second elastomer layer above these through openings, and by valve seats in the form of projections from the third elastomer layer of said base plate.