DE102007054309A1 - Highly dispersive matrix interaction length increasing method for determining concentration of e.g. blood, involves, involves detecting electromagnetic radiation, where amount of detected radiation is different from noise of detectors - Google Patents

Highly dispersive matrix interaction length increasing method for determining concentration of e.g. blood, involves, involves detecting electromagnetic radiation, where amount of detected radiation is different from noise of detectors

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Publication number
DE102007054309A1
DE102007054309A1 DE200710054309 DE102007054309A DE102007054309A1 DE 102007054309 A1 DE102007054309 A1 DE 102007054309A1 DE 200710054309 DE200710054309 DE 200710054309 DE 102007054309 A DE102007054309 A DE 102007054309A DE 102007054309 A1 DE102007054309 A1 DE 102007054309A1
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Prior art keywords
characterized
method
substance
device according
detection
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
DE200710054309
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German (de)
Inventor
Hansjörg Dr. Albrecht
Hans-Joachim Dipl.-Ing. Cappius
Jürgen Dr.rer.nat. Helfmann
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Laser und Medizin Technologie GmbH
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Laser und Medizin Technologie GmbH
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Priority to DE200710054309 priority Critical patent/DE102007054309A1/en
Publication of DE102007054309A1 publication Critical patent/DE102007054309A1/en
Application status is Withdrawn legal-status Critical

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Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
    • G01N21/17Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47Scattering, i.e. diffuse reflection
    • G01N21/4738Diffuse reflection, e.g. also for testing fluids, fibrous materials
    • G01N21/474Details of optical heads therefor, e.g. using optical fibres
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
    • G01N21/75Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/78Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
    • G01N21/84Systems specially adapted for particular applications
    • G01N21/8483Investigating reagent band
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
    • G01N21/17Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47Scattering, i.e. diffuse reflection
    • G01N21/4738Diffuse reflection, e.g. also for testing fluids, fibrous materials
    • G01N21/474Details of optical heads therefor, e.g. using optical fibres
    • G01N2021/4742Details of optical heads therefor, e.g. using optical fibres comprising optical fibres

Abstract

Method and apparatus for the high-sensitivity detection of a substance which is located on a visually strongly scattering solid substrate and which changes its optical properties when a second substance to be analyzed is added. In order to quantify even the lowest concentrations of this substance, the sensitivity according to the invention is increased by maximizing the path length. Thus, the interaction length is determined by the appropriate distance from Einstrahlort and detection site according to the invention under the constraints that the radiation does not leave the medium and the amount of radiation detected at the detector is sufficiently high for low-noise detection.

Description

  • task
  • On a (visually strongly scattering) solid substrate located Substance that adds its optical properties to a second substance to be analyzed, should be highly sensitive be detected to even the lowest concentrations of this substance yet to quantify.
  • State of the art
  • Under In certain circumstances, it is desired to be very low Concentrations of certain organic or inorganic compounds to eat. In medicine, for example, it is very useful the concentration of one, mostly in solution, given type of molecule, either from Nature from in physiological fluids (such as blood, saliva or Urine), or which are introduced into the living system was (such as drugs or toxins, or contaminants). by virtue of the rapidly progressing level of knowledge of the molecular basis both normal and morbid conditions more alive Systems, there is an increasing need for detection methods, which are quantitative, specific to the one of interest Molecule, highly sensitive and relatively easy to implement are. Examples of molecules of interest in one medical and / or biological context, but are not limited to medicines, intoxicants, Sexual and adrenaline hormones, biologically active peptides, circulating Hormones and antigens associated with tumors or infectious Agents. In the case of drugs, for example, requires the safe and effective use of a special drug, that its concentration in the circulatory system within relatively narrow Limits what is called the therapeutic area.
  • It It is known that the use of specific binding, radiation-emitting Particles can accomplish this task (radionuclide assay). disadvantage Such methods are the high cost of shielding the Radiation and the time-consuming disposal of such samples, and the technical complexity of generating short-lived radionuclides, the usually be used for binding to the analytes mentioned. In addition, contamination in the vicinity of the Measuring station make the entire measurement obsolete, as by the detectors used are the direction from which the radiation originates can not be distinguished well. In most cases Therefore, the use of the non-ionizing portion of the electromagnetic Radiation sought.
  • It is known that low concentrations of analyte in solutions can be detected optically spectroscopically with greater sensitivity if the optical path length through the sample is long. When the amount of sample is limited, as is often the case, the optimum measuring cell has a small cross-section and a long length. This uses for example the in EP 0 523 680 represented solution, where using a capillary tube, along the axis of which the liquid sample is irradiated, a small, non-scattering sample volume is detected highly sensitive.
  • It it is known that with the use of luminescent binding partners, which adhere to the substances to be detected and otherwise washed out a sensitive detection method exists, however through the additional application and washing steps a handling disadvantage with sources of error and high consumption has additives. In addition, the detection of a reliable darkened cell linked to the desired Sensitivity to realize. However, this conflicts with handling, because the single test elements should either be changed quickly (eg in a clinical laboratory) or by inexperienced people to be served.
  • An increasing number of sample formats (sample carriers) use a disposable test strip, a fluidic device or card. Conventional disposable test elements used in photometric tests usually take the form of the prior art test strips on which a test pad is applied. In addition, a control field can be applied, which checks the proper function (sample task). The test field receives the sample to be tested and typically contains reagents that are required for the test to be performed. The reagent system used in the test field can perform different functions. The sample is either applied directly to the test field or placed on a special field on the test strip and delivered to the test field. After a required reaction time characteristic color changes are measured reflection analysis by means of an analysis unit for the analysis of the sample. The evaluation device, which is intended to evaluate an analysis result, is usually suitable for a very specific type of test element of a particular manufacturer. The test elements and the evaluation device thus form mutually matched components and are commonly referred to collectively as an analysis system. Such analysis systems are used for example in the US 5,281,395 and US 5,424,035 described.
  • Different approaches to improve the Photometric measurement are already set forth in the prior art. In the DE 199 38 839 describes the problem of optical detection by the use of an optically transmissive medium with a reflective background, which aims at the use of common in the computer field CD-ROM readers as a measuring device. Due to the reflection, the detection layer is irradiated at least twice, which also causes an increase in sensitivity.
  • The representation of the prior art in the DE 102 10 436 A1 executes further examples:
    For example, the DE-A-692 27 545 an analysis method with measurement of the diffuse reflectance of individual frequency ranges. The light irradiated by means of a fiber optic is already diffusely reflected, depending on the wavelength and sample used, after short distances. The reflected light is then mixed with the same or wider fibers as in DE-A-42 42 083 . DE-A-44 15 728 . DE-A-43 37 570 and DE-A-199 34 038 described collected.
    [0004] The DE-A-43 37 570 discloses an arrangement which determines by determining the light transit time at which depth of the sample the light has been reflected. This additional information is inhomogeneous samples such. As the human skin valuable to gain a differentiated picture of the concentration distributions of individual analytes. A transmittance is as in WO-A-01/01852 is not possible for an IR absorption determination of glucose in human skin, because there is no sufficiently thin body site for it. The magneto-optical rotation of the polarization plane of polarized light in interaction with the molecules of a sample can be used to generate mixtures of e.g. B. to analyze hydrocarbons ( E. Hecht "Optics" Addison-Wesley, 1989 ). In this Faraday effect spectrometer, the sample is normally transilluminated (transmission spectrometer), with the wavelength of light within the sample as in JP-A-2000111585 and JP-A-63266323 described, constant and defined.
    The comparison of the observed absorption with absorption on samples of known concentration allows according to EP-A-94915814 . EP-A-91906149 . EP-A-552 300 and DE-A-199 63 561 Conclusions about their analyte concentration.
  • As soon as however, the liquid or solid sample to be analyzed scatters heavily (on average more than one scattering event on passage through the considered radiation-penetrated volume), this is Sample no longer reliable due to the scattered parts because the scattering does not hit the detector Radiation components are not differentiated from the absorbed beam components can be. Thus, in such samples is a simple Solution, like the above methods, no longer for Increase in sensitivity applicable.
  • Various solutions have been proposed to circumvent the problems of measurement on scattering media. The US 5,962,852 A describes a confocal-based method in which light is to be focused into different depths of a biological tissue in order to draw conclusions about the concentration of the analyte sought from the backscatter from the region of the focus. To help maintain the region of focus, it is proposed to use a low-coherence measurement method in addition to the confocal technique. Furthermore, it is proposed to use several wavelengths in order to obtain sufficient information about the absorption of the analyte. The determination is to be made via a calibration with a concentration series of the analyte. The technical complexity is enormous (n short-pulse emitter with x wavelengths and m time-dependent connected detectors arranged on a beam splitter). Also, the technical realization of the beam splitter is not clearly described, for which, however, at the same wavelength of the incident and detected light only the linear polarization comes into question, which is generally lost by scattering processes. Therefore, the detected proportion of the scattered light should be very low. In addition, with the depth of the focus region, the intensity of the light backscattered in the same way decreases sharply. Highly sensitive analysis will not be possible due to strong matrix scattering due to low light intensities at longer optical path lengths.
  • US 6,534,012 B1 describes a method which modulates the scattering by controlled compression of the test body or of the test fluid (the distance of the scattering centers is changed) and thus can detect the influence separately from the absorption which does not change due to the compression. However, this method is not applicable to a highly sensitive biological (analyte) contaminated substrate analyzer because of the possible cross contamination between successively measured single test elements.
  • at solid test substrates is the problem of increasing sensitivity only by concentration of the sample - which is often not reproducible and without distortions possible is - and by increasing the quota of chemical Reaction of the analytes on the substance bound in the test substrate to reach. Thus, the highly sensitive detection is only in tight Borders possible.
  • invention solution
  • The extension of the optical path length can be done in scattering media by irradiation at a defined location and the detection at an adjacent, at a defined distance (d) lying to point. The path length of the light (s) increases approximately linearly with the distance d ( RA Bolt, KR Rinzema, JJ ten Bosch, Pure Appl. Opt. 4, 1995 ). With small absorption, the extension of the path is s / d ≈ 4.5. As a result, in particular those portions of the radiation are detected, which come in the depth of the substrate by scattering at some distance back to the surface. Overall, this leads to an extension of the optical path traveled by the light and thus to the detection of a greater proportion of chemical conversion of the analytes to the substance bound in the test substrate.
  • The radiation distribution should be as completely as possible within the substrate. In the diffusion approximation of the radiation transport equation, the radiation distribution can be determined approximately as a function of the distance (d) of the locations for the irradiation and detection ( S. Feng, F.-A. Zeng, B. Chance, Appl. Opt. Vol 34 No 19, 1995 ). For the propagation of light, the attenuation coefficient increases μ eff = (3 μ a a + μ s (1 - g)) 1.2
  • The depth at which the maximum of the detected radiation distribution lies reaches a maximum in the middle between the irradiation site and the detection site. This depth can be determined for two cases: strong (μ eff d >> 1) and weak ((μ eff d << 1)) attenuation to: z Max = (2) 1.2 d / 4 weak damping z Max = (d / (2 μ eff )) 1.2 strong damping
  • These solutions are in 2 for a typical attenuation of μ eff = 0.5 mm -1 .
  • In order to keep the radiation distributions as completely as possible in the substrate, z max should be equal to or less than half the substrate thickness.
  • The Measuring principle works in the same way regardless of whether the irradiation and detection on the same side of the substrate or lying on opposite sides.
  • For technical implementation, the following points should be considered:
    Due to the displacement of the optical path into the depth of the substrate, a chemical conversion of the analyte must also be ensured in the depth of the substrate, or its active penetration depth should be taken into account when choosing the distances between the irradiation site and the detection site.
  • By the scattering reduces the proportion of the radiation intensity, which is to be detected at the detection site. Therefore, depending on of the test substrate, the absorption properties of the on a (optically strongly scattering) solid substrate bound substance, their optical properties when adding a second too altered substance (in the following color reaction called, but not limited to the visible area) and the parameter of the detector is the distance beam location to the detection location and the Einstrahlintensität be chosen appropriately.
  • In Continuation of the inventive idea, it is possible the geometry of the illumination and detection surfaces adjust the specifics given by the test reaction. Especially can be at a linear or rectangular shape of the test substrate bound substance the shape of the surfaces be chosen so that through the optical integration over the lighting surface noise is minimized. to Clarification is as a possible execution the design of a rectangular detector and a linear one Called lighting, the irradiated volume the entire Width of the test substrate / sample carrier includes and not limited to an example circular area is, which lead to a lack of utilization of the edge areas would. Without limitation of the invention are also other conceivable arrangements, such as circular or three-dimensional Designs of the test substrate included.
  • Also in continuation of the inventive concept, the detection a surface not intended for the color reaction of the test substrate / slide, for improved zero balance to achieve and above that the sensitivity and Increase robustness of the measurement.
  • Also in continuation of the inventive concept can be attached to two or more spatially separated locations on the sample a different concentration of the tightly bound first substance made the comparison more robust the measurement and make it more sensitive. For this purpose, the task is symmetrical between the test fields, with the analyte moving in all directions moved evenly to the test fields.
  • Also in continuation of the inventive concept, a plurality of similar tests can be performed in parallel on the substrate in spatially separated locations, the test result is sequentially detected by one of the described light source detector assemblies or partially or completely parallel in time by a plurality of be written light source detector arrangements is detected.
  • Description of the drawings
  • 1 shows the principle of increasing the interaction length by way of example on a fiber-coupled solution for a reflection arrangement. In the free jet, ie with a light source which illuminates the substrate directly or via optics, and as a transmission arrangement, ie light source and detector or their respective fibers are located on opposite sides of the substrate, the principle is equally applicable.
  • 2 shows the achievable depth of the irradiated volume as a function of the distance between the irradiation site and detection site with 2 different strong scattering test elements.
  • 3 shows a result of a chromatography assay in which various evaluations of the color reaction zone have been made. The rows marked C were illuminated and measured at the same position (local reflection). In the row B was illuminated on a small area and detected on a large, the illumination surface containing area.
  • In the row A was illuminated on a small area and spatially separated from it on a small area detected. All images are set in brightness and contrast, that in each case the full extent of the 8 bit recording (scan of illumination area and detection surface over the test strip with, for example a laser scan microscope) is exploited. The variations in of a series include the variation of the lateral Distance of the illumination surface from the detection surface (only row A), the size change the illumination / detection area and the use of crossed polarizers for illumination and detection radiation.
  • 4 shows the profiles from the pictures in 3 where all pixels of the vertical (each column) were averaged and averaged over groups of 8 pixels horizontal (lines) of images. The determined signal to noise ratio (signal / noise) is indicated next to the profiles. It can be clearly seen that at least one increase in sensitivity is expected to be five times greater if, instead of local reflection, the reflection is measured laterally.
  • 1
    Einstrahlfaser
    2
    detection fiber
    3
    second Position of the detection fiber
    4
    strongly scattering matrix with reagent system
    5
    carrier substrate
    6
    By Scattering radiated area of the photons detected in FIG. 2
    7
    By Scattering radiated area of the detected in 3 photons
  • QUOTES INCLUDE IN THE DESCRIPTION
  • This list The documents listed by the applicant have been automated generated and is solely for better information recorded by the reader. The list is not part of the German Patent or utility model application. The DPMA takes over no liability for any errors or omissions.
  • Cited patent literature
    • - EP 0523680 [0004]
    • - US 5281395 [0006]
    • US 5424035 [0006]
    • - DE 19938839 [0007]
    • DE 10210436 A1 [0008]
    • - DE 69227545 A [0008]
    • - DE 4242083 A [0008]
    • - DE 4415728 A [0008]
    • - DE 4337570 A [0008, 0008]
    • - DE 19934038 A [0008]
    • WO 01/01852 A [0008]
    • - JP 2000111585 A [0008]
    • - JP 63266323A [0008]
    • - EP 94915814 A [0008]
    • - EP 91906149 A [0008]
    • - EP 552300 A [0008]
    • - DE 19963561 A [0008]
    • - US 5962852 A [0010]
    • US 6534012 B1 [0011]
  • Cited non-patent literature
    • - E. Hecht "Optics" Addison-Wesley, 1989 [0008]
    • - RA Bolt, KR Rinzema, JJ ten Bosch, Pure Appl. Opt. 4, 1995 [0013]
    • - S. Feng, F.-A. Zeng, B. Chance, Appl. Opt. Vol 34 No 19, 1995 [0014]

Claims (19)

  1. Method and apparatus for increasing the interaction length in a strongly scattering matrix characterized in that - electromagnetic radiation is detected at a lying at a defined distance to the Einstrahlort detection site, which on interaction with a bound in a strongly scattering matrix first substance conclusions about the amount of detecting second substance allows - the distance is selected defined by a calculation rule that the interaction length is maximum and the detected with suitable detectors amount of electromagnetic radiation is to be distinguished from the noise of the detector taking into account the achievable over the first substance changes to the electromagnetic radiation which is triggered by the second substance present quantitatively at the detection limit.
  2. Method and device according to claim 1, characterized characterized in that said bound first substance by Reaction with the second substance to be analyzed an absorption change generates the electromagnetic radiation.
  3. Method and device according to claim 2, characterized characterized in that said bound first substance is the lag the second substance to be analyzed prevents and thus an absorption change generates the electromagnetic radiation.
  4. Method and device according to claim 2, characterized characterized in that said bound first substance by a reaction of reaction with the second substance to be analyzed an absorption change of the electromagnetic radiation generated.
  5. Method and device according to claim 1, characterized characterized in that said bound first substance by Reaction with the second substance to be analyzed if appropriate Illumination with electromagnetic radiation changed one Scattering generated.
  6. Method and device according to claim 2 or 5, characterized in that said reaction is more than two substances is generated.
  7. Method and device according to claim 1, characterized characterized in that the detection of the electromagnetic radiation by suitable pulsed or modulated operation of light source and detector (lock-in technique) of disturbing influences can be separated.
  8. Method and device according to claim 1, characterized characterized in that the detection of the electromagnetic radiation by placing a suitable filter in front of the detector of spurious radiation unaffected.
  9. Method and device according to claim 1, characterized characterized in that the detection of the electromagnetic radiation by detecting a background signal of disturbing influences is cleaned up.
  10. Method and device according to claim 1, characterized characterized in that Einstrahlort and detection site on the same Side of the test strip lie (reflection arrangement).
  11. A method and apparatus according to claim 10, characterized in that the calculation rule for the distance d of the illumination and detection surfaces at high attenuation (μ eff d >> 1) is: z max ≤ ½ substrate thickness, with z max = (d / (2 μ eff )) 1/2 .
  12. A method and apparatus according to claim 10, characterized in that the calculation rule for the distance d of the illumination and detection surfaces at low attenuation (μ eff d << 1) is: z max ≤ ½ substrate thickness, with z max = (2) 1 / 2 d / 4.
  13. Method and device according to claim 1, characterized characterized in that Einstrahlort and detection location on different Pages of the test strip lie (transmission arrangement).
  14. A method and apparatus according to claim 13, characterized in that the calculation rule for the distance d of the illumination and detection surfaces at high attenuation (μ eff d >> 1) is: z max ≤ ½ substrate thickness, with z max = (di (2 μ eff )) 1/2 .
  15. Method and device according to claim 13, characterized in that the calculation rule for the distance d of the illumination and detection surfaces at low attenuation (μ eff d << 1) is: z max ≤ ½ substrate thickness, with z max = (2) 1 / 2 d / 4.
  16. Method and device according to one of the claims 10 or 13, characterized in that the shape of the illumination surface and detection surface so to the surface of the substance to be detected adjusted, that a spatial averaging is achieved.
  17. Method and device according to one of claims 10 or 13, characterized in that an array of a plurality of light sources or a plurality of detectors to the surface of the substance to be spatially averaged.
  18. Method and device according to one of the claims 10 or 13, characterized in that a plurality of test fields spatially are arranged separately on the substrate and temporally sequential be scanned.
  19. Method and device according to one of the claims 10 or 13, characterized in that a plurality of test fields spatially are arranged separately on the substrate and by a plurality of light source detector assemblies be scanned parallel in time.
DE200710054309 2007-11-08 2007-11-08 Highly dispersive matrix interaction length increasing method for determining concentration of e.g. blood, involves, involves detecting electromagnetic radiation, where amount of detected radiation is different from noise of detectors Withdrawn DE102007054309A1 (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102014003470A1 (en) 2014-03-07 2015-09-10 Laser- Und Medizin-Technologie Gmbh, Berlin Sensor device for spatially resolving detection of target substances
DE202014010964U1 (en) 2014-03-07 2017-03-10 Laser- Und Medizin-Technologie Gmbh, Berlin Sensor device for spatially resolving detection of target substances

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