Autonomous Calibration for Optical Analysis System

Abstract

The present invention provides an autonomous calibration of a multivariate based spectroscopic system that is preferably implemented as a multivariate based spectrometer. The spectroscopic system is based on a multivariate optical element that provides a spectral weighting of an incident optical signal. Spectral weighting is performed on the basis of spatial separation of spectral components and subsequent spatial filtering by means of a spatial light modulator. Calibration of the spectroscopic system is based on a dedicated calibration segment of the spatial light modulator, whose position corresponds to a characteristic calibration or reference wavelength of the incident optical signal. Preferably, the calibration or reference wavelength is given by the wavelength of the excitation radiation generated by the optical source that serves to induce scattering processes in a volume of interest.

Description

In the following preferred embodiments of the invention will be described in detail by making reference to the drawings in which:

FIG. 1 is a schematic diagram of an embodiment of a blood analysis system,

FIGS. 2 a and 2b are spectra of the optical signal generated from blood in the skin and from a sample comprising one anlayte in a solution,

FIG. 3 is a spectral weighting function implemented in a multivariate optical element,

FIG. 4 schematically illustrates a block diagram of the optical analysis system,

FIG. 5 schematically illustrates a block diagram of possible detector configurations,

FIG. 6 shows a perspective illustration of the spatial light modulating mask and a corresponding detector,

FIG. 7 shows a flowchart of calibrating the optical analysis system.

In the embodiment shown in FIG. 1 the spectroscopic system is schematically illustrated. The spectroscopic system has an optical analysis system 20 for determining an amplitude of a principal component of an optical signal. The spectroscopic system further has a light source 1 for providing light for illuminating a sample 2 comprising a substance having a concentration and thereby generating the principal component. The amplitude of the principal component relates to the concentration of the substance. The light source 1 is a laser such as a gas laser, a dye laser and/or a solid state laser such as a semiconductor or diode laser. The optical analysis system 20 is part of a blood analysis system 22. The blood analysis system further comprises a computational element 19 for determining the amplitude of the principal component, hence for determining the composition of the compound. The sample 2 comprises skin with blood vessels. The substance may be one or more of the following analyses: glucose, lactate, cholesterol, oxy-hemoglobin and/or desoxy-hemoglobin, glycohemoglobin (HbAlc), hematocrit, cholesterol (total, HDL, LDL), triglycerides, urea, albumin, creatinin, oxygenation, pH, bicarbonate and many others. The concentrations of these substances is to be determined in a non-invasive way using optical spectroscopy. To this end the light provided by the light source 1 is sent to a beam splitter 3 which reflects the light provided by the light source towards the blood vessels in the skin. The light may be focused on the blood vessel using an objective 12. The light may be focused in the blood vessel by using an imaging and analysis system as described in the international patent application WO 02/057759.

By interaction of the light provided by the light source 1 with the blood in the blood vessel an optical signal is generated due to Raman scattering and fluorescence. The optical signal thus generated may be collected by the objective 12 and sent to the dichroic mirror 3. The optical signal has a different wavelength than the light provided by the light source 1. The dichroic mirror is constructed such that it transmits at least a portion of the optical signal.

A spectrum 100 of the optical signal generated in this way is shown in FIG. 2A. The spectrum comprises a relatively broad fluorescence background (FBG) 102 and relatively narrow Raman bands (RB) 104, 106, 108. The x-axis of FIG. 2A denotes the wavelength shift with respect to the 785 nm of the excitation by light source 1 in wave numbers, the y-axis of FIG. 2A denotes the intensity in arbitrary units. The x-axis corresponds to zero intensity. The wavelength and the intensity of the Raman bands, i.e. the position and the height, is indicative for the type of analyte as is shown in the example of FIG. 2B for the analyte glucose which was dissolved in a concentration of 80 mMol in water. The solid line 110 of FIG. 2B shows the spectrum of both glucose and water, the dashed line 112 of FIG. 2B shows the difference between the spectrum of glucose in water and the spectrum of water without glucose. The amplitude of the spectrum with these bands is indicative for the concentration of the analyte.

Because blood comprises many compounds each having a certain spectrum which may be as complex as that of FIG. 2B, the analysis of the spectrum of the optical signal is relatively complicated. The optical signal is sent to the spectroscopic system 20 according to the invention where the optical signal is analyzed by a MOE which weighs the optical signal by a weighting function shown e.g. schematically in FIG. 3. The weighting function of FIG. 3 is designed for glucose in blood. It comprises a positive part P and a negative part N. The positive part P and the negative part N each comprise in this example more than one spectral band.

Here and in the remainder of this application the distance between a focusing member and another optical element is defined as the distance along the optical axis between the main plane of the focusing member and the main plane of the other optical element.

A computational element 19 shown in FIG. 1 is arranged to calculate the difference between the positive and negative signal. This difference is proportional to the amplitude of the principal component of the optical signal. The amplitude of the principal component relates to the concentration of the substance, i.e. of the analyte. The relation between the amplitude and the concentration may be a linear dependence.

FIG. 4 schematically illustrates a block diagram of the blood analysis system 22 and the optical analysis system 20 in a more detailed way. Here, the optical analysis system 20 is shown as an integral part of the blood analysis system 22. However, the invention is by no means restricted to blood analysis but may be universally applied to various spectroscopic operational areas. In the illustrated embodiment, the optical analysis system 20 has a dispersive optical element 30, a lens 32, a spatial light transmission mask 34 and two detectors 40, 42. The output of the two detectors 40, 42 is coupled into a control unit 60 that is in turn adapted to manipulate the path of the optical beams inside the optical analysis system 20 as well as to calibrate or to tune the light source 1.

The light source 1 is preferably implemented as a laser light source operating in the near infrared spectral range. The light source produces an optical beam of excitation radiation 50 that is directed towards the beam splitter 3. The arrangement of beam splitter 3 and objective lens 12 serves to focus the excitation radiation 50 into the volume of interest 4 of the biological sample 2. Inside the volume of interest the excitation radiation typically induces a variety of scattering processes. In particular inelastic scattering processes, like Stokes or Anti-Stokes scattering processes lead to a Raman spectrum that allows for spectral analysis for determination of the composition of the volume of interest 4. Typically, a part of the Raman spectrum is back-scattered and re-enters the objective lens 12 in a counter propagating way with respect to the excitation radiation 50. The collected return radiation is then transmitted through the beam splitter 3 and becomes incident on the dispersive optical element 30.

The dispersive optical element 30 can be implemented by e.g. a transmission or reflection grating, a prism or any other dispersive element that provides spatial separation of spectral components of an incident light beam. The dispersive element 30 provides spatial separation of the spectral components of the incident return radiation in the horizontal direction, as shown in FIG. 4. The lens 32 provides focusing of the various spectral components 52, 54 of the return radiation to different positions on the spatial light transmission mask 34. Depending on the light transmission pattern of the spatial light transmission mask 34, the various spectral components of the return radiation are selective attenuated or even blocked. In this way a spectral weighting function as required for multivariate optical analysis can be effectively realized.

For an accurate and reliable determination of a concentration of a particular compound of the volume of interest 4, the relative position between the evolving spectrum and the spatial light transmission mask 34 is of high relevance. Already a slight mismatch between the position of the spectrum and the horizontal alignment of the transmission mask 34 may lead to an insufficient result of the optical and spectral analysis of the return radiation.

The transmission mask 34 comprises a slit 36 at a first position that serves as a reference segment. When properly aligned the position of the reference segment 36 corresponds to the position of the spectral component 52 of the return radiation. This spectral component 52 serves as a reference line in the spectrum. Preferably, the reference line is determined by the wavelength of the excitation radiation 50. Typically, the spectrum not only comprises spectral components from inelastic but also from elastic scattering processes that leave the wavelength of the optical radiation substantially unaffected during a scattering process. Moreover, elastically scattered radiation may even represent a major portion of the return radiation. The slit 36 is therefore highly transmissive for the wavelength of the excitation radiation. Hence, radiation that is transmitted through the slit 36 is detected by means of the detector 40 that is adapted to generate a calibration signal that is fed into the control unit 60.

Slit 38 of the transmission mask 34 is located at a different horizontal position compared to slit 36. It therefore serves to attenuate and/or to transmit a spectral component of the return radiation featuring a different wavelength than the excitation radiation 50. Typically, the transmission mask 34 comprises a plurality of slits 38 that are located at different horizontal locations on the transmission mask 34. The slits 36, 38 might be implemented as slit apertures. Additionally, they might be combined with neutral density filters for selectively attenuating different spectral components of the return radiation to various degrees.

The transmission mask 34 can be implemented as a fixed transmission mask providing a single spatial transmission pattern. In such a case the optical analysis is dedicated for determining the concentration of a single particular compound in the volume of interest 4. Various compounds or analytes in the volume of interest 4 might be investigated and spectrally analyzed by replacing the transmission mask 34 by another compound specific transmission mask that is dedicated for multivariate optical analysis of a different compound.

Alternatively, the transmission mask 34 might be implemented as a reconfigurable transmission mask. This can for example be realized by implementing the transmission mask 34 on the basis of a liquid crystal cell in combination with a crossed polarizer arrangement.

Detector 42 serves to detect radiation that has been transmitted through the transmission mask 34. Preferably, some kind of focusing means is placed between transmission mask 34 and detector 42 in order to collect a plurality of transmitted spectral components onto the detector 42. Hence, detector 42 generates an output signal in response to detect a plurality of transmitted spectral components. The output signal of detector 42 is therefore directly indicative of the concentration of a particular compound that can be calculated by means of the computational element 19.

The calibration signal generated by means of detector 40 is provided to the control unit 60 that in turn is adapted to manipulate the horizontal position of the transmission mask 34, to shift or to rotate the dispersive optical element 30 or to tune the light source 1. Whenever the light intensity detected by detector 40 does not correspond to an intensity that is expected for an accurate calibration of the optical analysis system, the control unit may successively modify the position or orientation of the optical components 30, 34 or may tune the light source 1 until the intensity detected by detector 40 corresponds to an expected value. Additionally, the control unit 60 may perform a position scan of transmission mask 34 in order to maximize the light intensity detected by detector 40.

In this way the intrinsically present excitation radiation component of the collected return radiation can be effectively exploited for calibration of the optical analysis system 20.

FIG. 5 shows an alternative arrangement of the two detectors 40, 42 with respect to the spatial light transmission mask 34. Here, detector 40 that is adapted for detection of the calibration signal 52 is positioned directly behind the slit 36 of the transmission mask 34. In this way no additional optical means, like lenses are required for directing the transmitted radiation onto the detector 40. Additionally, by mechanically fixing or mechanically coupling detector 40 to the transmission mask 34, the detector 40 follows any movement of the transmission mask 34.

Moreover, FIG. 5 also depicts a collection lens 56 that serves to focus transmitted spectral components of the return radiation 54 onto the detector 42 for multivariate optical analysis.

FIG. 6 shows a perspective illustration of the transmission mask 34 and the detector 40. The transmission mask 34 features three slit apertures 36, 38, 39 that serve to transmit a corresponding spectral component of the return radiation. In particular, slit aperture 36 serves as reference segment of the transmission mask 34 and is therefore horizontally positioned in such a way that it provides transmission of an excitation radiation spectral component of the return radiation 52.

Detector 40 is implemented as a split detector that has at least two detection segments 62, 64. In this way the detector 40 not only allows to determine whether the optical analysis system is not properly calibrated but also provides information of a type of positional mismatch between the spatial distribution of the spectrum and the transmission mask 34. For example, when the two detection segments 62, 64 of the detector 40 detect equal transmitted intensity, this gives a clear indication that the optical analysis system is accurately calibrated. Whenever one of the two detection segments 62, 64 detects a larger intensity than the other one, information whether the transmission mask 34 has to be shifted to the left or to the right is directly obtained. In this way a calibration of the optical analysis system can be performed in a less time intensive way, i.e. a scan for a maximum of transmitted intensity has in principle only to be performed along one direction.

FIG. 7 illustrates a flowchart for performing the inventive calibration method making use of e.g. a photodiode as detector 40. In a first step 700 radiation that is transmitted through the reference segment 36 is detected. This detected light intensity is stored in a subsequent step 702 as intensity I_x. When no reference intensity is stored in the control unit that allows to determine whether the measured intensity I_x corresponds to the maximum intensity, thereby indicating that the optical analysis system is accurately calibrated, in a subsequent step 704 a system parameter of the optical analysis system is modified. Modification of a system parameter typically refers to translating of the spatial transmission mask 34, translating or rotating of the dispersive optical element 30, tuning or calibrating of the laser light source 1 and reconfiguring of the spatial light transmission pattern of the transmission mask 34.

Modification of these system parameters either corresponds to a successive modification of a single system parameter or to simultaneous or combined modification of a variety of system parameters. After or during modification of those system parameters performed in step 704 radiation that is transmitted through the reference segment is repeatedly detected in step 706. A corresponding intensity referred to as I_x+1 is stored in step 708. Thereafter, the detected light intensities I_x and I_x+1 are compared in step 710. Comparison of the two light intensities that correspond to slightly different calibration configurations of the optical analysis system typically refers to a comparison of their absolute value.

In case that the successively detected intensity of I_x+1 is larger than the previously detected intensity I_x the method continues with step 714 where intensity I_x is replaced by the recently detected intensity I_x+1. In this way the intensity I_x principally refers to a temporary maximum detected intensity. After step 714 the method returns to step 704 where again a system parameter of the optical analysis system is modified.

In the other case, where the recently detected intensity I_x+1 is not larger than the previously detected intensity I_x, after step 710 the method continues with step 712, where the last modification performed in step 704 is undone. In this case the modification of a system parameter performed in 704 did not lead to an improvement of the optical analysis system's calibration. Therefore, the performed modification is cancelled. After this canceling performed in step 712 the method returns to step 704, where another system parameter is preferably subject to modification.

In this way the optical analysis system is iteratively and constantly calibrated by means of seeking for a maximum of transmitted intensity of the reference spectral component.

Consequently, the invention provides an autonomous calibration of a multivariate based spectrometer. By making use of the excitation radiation as calibration or reference line in the obtained spectrum, no additional light source for calibration of the optical analysis system is needed.

LIST OF REFERENCE NUMERALS

• 1 light source
• 2 sample
• 3 beam splitter
• 4 volume of interest
• 12 objective
• 19 computational element
• 20 optical analysis system
• 22 blood analysis system
• 30 dispersive element
• 32 lens
• 34 spatial light transmission mask
• 36 slit
• 38 slit
• 40 detector
• 42 detector
• 50 excitation radiation
• 52 return radiation
• 54 return radiation
• 56 lens
• 60 control unit
• 62 detection segment
• 64 detection segment

Claims

1. A spectroscopic system for determining a principal component of an optical signal, the optical signal comprising return radiation from a volume of interest (4), the spectroscopic system comprising: light source (1) for generating an excitation radiation, the excitation radiation (50) being adapted to be transmitted into the volume of interest,an objective (12) for collecting return radiation from the volume of interest,a dispersive optical element (30) for spatially separating spectral components of the return radiation in a first direction,spatial light manipulation means (34) for modulating the spectral components of the return radiation, the spatial light manipulation means further having a reference segment (36) at a first position, the reference segment being at least partially transparent for the excitation radiation,at least a first detector (40) for detecting radiation being transmitted through the reference segment,a control unit (60) being adapted to calibrate the optical analysis system on the basis of the radiation being detected by means of the at least first detector.

2. The spectroscopic system according to claim 1, wherein the reference segment (36) comprises a slit aperture and the first position substantially corresponds to the wavelength of the excitation radiation.

3. The spectroscopic system according to claim 1, wherein the control unit (60) is adapted to translate the spatial light manipulation means (34) along the first direction.

4. The spectroscopic system according to claim 1, wherein the control unit (60) is adapted to control the light source (1) in order to modify the wavelength of the excitation radiation (50).

5. The spectroscopic system according to claim 1, wherein the control unit (60) is adapted to rotate or to translate the dispersive optical element (30).

6. The spectroscopic system according to claim 1, wherein the spatial light manipulation means (34) are modifiable, the control unit (60) being further adapted to modify the spatial light manipulation means.

7. The spectroscopic system according to claim 1, wherein the at least first detector (40) comprises a segmented detector having at least two detector segments (62, 64) being separated along the first direction.

8. The spectroscopic system according to claim 1, wherein the at least first detector (40) being integrated into the spatial light manipulation means (34) at the first position.

9. A method of calibrating a spectroscopic system having a light source (1) for generating an excitation radiation (50) being adapted to be transmitted into a volume of interest (4), an objective (12) for collecting return radiation from the volume of interest, a dispersive optical element (30) for spatially separating spectral components of the return radiation in a first direction and spatial light manipulation means (34) for modulating the spectral components of the return radiation, the method of calibrating comprising the steps of: detecting radiation being transmitted through a reference segment (36) of the spatial light manipulation means by means of an at least first detector (40), the reference segment being at least partially transparent for the excitation radiation and being located at a first position on the spatial light manipulation means,calibrating the spectroscopic system on the basis of radiation being detected by means of the at least first light detector.

10. The method according to claim 9, wherein calibrating of the spectroscopic system comprising maximizing the radiation being transmitted through the reference segment.

11. The method according to claim 9, wherein calibrating of the spectroscopic system further comprising the steps of: translating the spatial light manipulation means along the first direction, and/ormodifying the wavelength of the excitation radiation by means of controlling the light source, and/orrotating and/or translating the dispersive optical element, and/orreconfiguring a spatial transmission pattern of the spatial light manipulation means.

12. A computer program product for calibrating a spectroscopic system, the spectroscopic system having a light source (1) for generating an excitation radiation (50) being adapted to be transmitted into a volume of interest (4), an objective (12) for collecting return radiation from the volume of interest, a dispersive optical element (30) for spatially separating spectral components of the return radiation in a first direction and spatial light manipulation means (34) for modulating the spectral components of the return radiation, the computer program product comprising computer program means being adapted to: (a) store a first output signal of an at least first detector (40), the first output signal being generated in response to detect a radiation being transmitted through a reference segment (36) of the spatial light manipulation means,(b) store a second output signal of the at least first detector after a modification of the position of the spatial light manipulation means along the first direction and/or after a modification of the wavelength of the excitation radiation and/or after a modification of the orientation and/or position of the dispersive optical element and/or after a reconfiguration of a spatial transmission pattern of the spatial light manipulation means,(c) compare the first and second output signal of the at least first detector,(d) repeat steps (a) to (c) until the output signal is indicative of a maximum of radiation being detected by means of the at least first detector.