WO2010095071A1 - Sensing device for detecting a target substance - Google Patents
Sensing device for detecting a target substance Download PDFInfo
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- WO2010095071A1 WO2010095071A1 PCT/IB2010/050553 IB2010050553W WO2010095071A1 WO 2010095071 A1 WO2010095071 A1 WO 2010095071A1 IB 2010050553 W IB2010050553 W IB 2010050553W WO 2010095071 A1 WO2010095071 A1 WO 2010095071A1
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- region
- investigation
- reference region
- sensing device
- light
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/55—Specular reflectivity
- G01N21/552—Attenuated total reflection
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration
- G01N21/274—Calibration, base line adjustment, drift correction
Definitions
- the present invention relates to a sensing device and an analyzing device for sensing a target substance in an investigation region.
- the invention further relates to a corresponding sensing method for sensing a target substance in an investigation region.
- US 7,317,534 B2 provides a measuring method comprising a measuring unit with a film layer having a detecting area where a target molecule is fixed to the surface thereof and a reference area where no ligand is fixed to the surface thereof.
- a photo detector detects the intensities of light beams reflected in total internal reflection at the detecting area and reference area, respectively. Further, the result of measurement in the detecting area is calibrated on the bases of the result of measurement in the reference area.
- US2005/0052655 Al describes an interferometer comprising an optical body adapted in operation to mount a measurement area comprising a film which is capable of acting as a two dimensional environment for surface plasmons and an adjacent reference area, an optical beam generation means for irradiating the reference and measurement areas with radiation capable of generating surface plasmon resonance, optical means for combining radiation reflected from the reference and measurement areas, and pixelated detection means for generating data representing two dimensional images of the combined radiation beams.
- a sensing device for detecting a target substance in an investigation region comprising
- the invention is based on the idea that a signal corresponding to the presence of a substance at the investigation region obtained by detecting light reflected from the investigation region under total internal reflection conditions can be calibrated by a signal corresponding to a standard reflecting light under total internal reflection conditions.
- a reference element effectively shielding the reference region from the influence of substances potentially present at the investigation region allows to measure a property, typically the intensity, of light reflected at the reference region in a standardized form, i.e. not dependent on the presence or absence of the substance to be analysed.
- the light reflected at the reference region is substantially only a function of factors other than the presence or absence of the substance and thus reflects errors due to measurement errors, e.g. increases in temperature or fluctuations in the light output reaching the reference region.
- light reflected at the reference region under total internal reflection conditions is considered unaffected by the presence or absence of the substance when a selected property of the light, typically its intensity, is not altered by the presence or absence of the substance by more than 10 %, preferably not more than 1 %, and most preferably not more than 0.1 %. Particularly preferred is a range of 0.05 to 0.3%.
- the reference element shields the reference region not only from the substance to be analysed, but also against other substances potentially influencing an evanescent field present at the reference region.
- This advantageously allows to analyse the presence of a substance in a coloured medium, as the coloration of the medium would not interfere with the light reflected at the reference region under total internal reflection conditions.
- the reference region can serve as a reliable standard.
- the method of the present invention and the sensing devices of the present invention can also advantageously be used to complement analytical methods involving measurement of transmitted light, for example measurement of light absorption and optical density. It is thus advisable for the reference element to be preferably solid, gel-like or otherwise resistant against being washed away by the medium.
- total internal reflection shall include the case frequently termed “frustrated total internal reflection", where some of the incident light is lost during the reflection process.
- the reflected light beam originating at the investigation region will typically consist of or comprise light of the incident light beam that was totally internally reflected at the investigation region of the sensing surface, which might be a binding surface for binding the target substance. It may, however, also comprise light from other sources like a fluorescence stimulated in the investigation region.
- the refractive index and dimensions of the reference element are preferably chosen such that the dimensions exceed the exponential decay length of an evanescent field elicited therein, i.e. such that the evanescent field is substantially not located outside of the reference element and of a carrier on which the reference element is located.
- the thickness and lateral dimensions of the reference element are then chosen in such a way that the evanescent field generated under total internal reflection conditions inside the reference element is substantially damped out for a chosen wavelength and angle of incidence of light.
- the angle ⁇ CTlt defines the critical angle.
- the lateral shape of the reference element can be chosen arbitrarily, thus allowing the present device to be adapted to a plethora of ancillary conditions.
- the reference element is a film or layer having a thickness of at least
- Such elements advantageously can be included in microsensors allowing to analyse minute quantities of a medium.
- the investigation region comprises a binder for binding a target.
- the target can be a target substance which can influence an evanescent field on its own.
- analyte the target would be such label.
- the target will then be a substance, preferably a magnetic bead, comprising a coupling section to attach to the analyte, preferably by an attachment specific for the analyte in view of other substances that are expected to be further comprised in the medium.
- the coupling section may be linked covalently or non-covalently to the analyte and the rest of the target substance, respectively.
- the target comprises one, two, three or more antibodies or Fab fragments thereof, including F(ab') 2 fragments, which can bind to the analyte at the same or at different sections of the analyte.
- the investigation region may comprise one, two or more types of binders.
- the types of binders can be specific for different target molecules or for different sections and antigens of one or more target molecules.
- the sensing surface of the present invention is thus suitable for different kinds of biosensors and measuring methods.
- Another type of assay for determining the concentration of an analyte in a medium can be a competitive binding assay.
- quantitation of an analyte concentration can be achieved by analysing the presence or absence of a target at a respective investigation region as a result of a competition between the analyte and an analyte-like substance for binding by the binder and/or the target or its respective coupling section.
- the sensing device of the present invention is preferably adapted to analyse the presence of a substance (i.e. target or, if applicable, analyte) in a medium at a concentration of less than or equal to 1 nM, even more preferably from 1 to 1000 pM and most preferably from 10 to 1000 fJVl.
- a substance i.e. target or, if applicable, analyte
- concentration of less than or equal to 1 nM even more preferably from 1 to 1000 pM and most preferably from 10 to 1000 fJVl.
- Such low concentrations typically require long measuring times and the obtained signals from the medium are small as well.
- the signals generated by typical optical sensor devices can drift over time with no changes in assay composition.
- the light output of the light source may vary, e.g. due to temperature changes of the environment or within the biosensor device. The drift can lead to significant deviations of the obtained signal compared to the real amount of target molecules bound at the investigation region.
- the reference region is preferably adjacent to the investigation region. This way, errors in intensity measurements of light reflected under total internal reflection conditions e.g. due to slight variations in the carrier material compositions can be minimized.
- a reference region is considered adjacent when it is separated from an investigation region, preferably an investigation region having attached a binder thereto as described above, by at most 5 mm, more preferably 0.5 to 1 mm, and most preferably 0.1 to 0.5 mm.
- the material of the reference element can be chosen arbitrarily, as long as it allows total internal reflection at the reference region for a preselected wavelength and angle of incidence of light and sufficiently shields the reference region as indicated above.
- the reference element is a solid, even more preferably the reference element is a film.
- glasses are considered solids.
- Further preferred materials of the reference element are chosen from polymers, bio molecules and particularly proteins, nucleic acids and polysaccharides, gels, sol-gels and other plastics.
- the reference region comprises a mirror to reflect incident light.
- a mirror to reflect incident light.
- Such mirror frequently termed true mirror, can be applied in the form of a dielectric multilayer or metallic coating and is particularly suitable as a reference element in such conditions where finding a reference element material with sufficiently low refractive index is difficult.
- the sensing surface will comprise two or more investigation regions at which different incident light beams can be totally internally reflected.
- One device then allows the processing of several investigation regions and thus for example the search for different target substances, the observation of the same target substances under different conditions and/or the sampling of several measurements for statistical purposes.
- the "different incident light beams" may optionally be components of one broad light beam that is homogeneously generated by one light source, they may be individual separate light beams addressing the investigation regions and/or reference regions simultaneously (optionally through the same or through different optical windows), and/or they may be temporally different (i.e. be generated by one generic light beam scanning the investigation regions).
- the "different incident light beams" are part of one broad light beam simultaneously illuminating the one, two or more reference region(s) and the one, two or more investigation region(s).
- a detector may then measure the light reflected from the respective regions separately, als will be described in greater detail below.
- the sensing device preferably is a cartridge having a carrier comprising the sensing surface thereon. Such cartridges can be advantageously used with analyzing devices, to adapt the analyzing devices to specific measurement tasks.
- the sensing device of the present invention preferably further comprises
- a light source to direct incident light to the investigation region and the reference region of the sensing surface such that the incident light is reflected under total internal reflection conditions at the investigation region and the reference region thereby generating reflected light
- Such device makes use of the advantage of the present invention, i.e. the sensing device allows to calibrate the first characteristic signal in view of the second characteristic signal.
- the sensing device thus particularly facilitates reliable, significant measurements, limits measurement errors and allows detecting a target substance at the investigation region(s) at low concentrations by enabling long(er) measurement times.
- the calibrator is preferably adapted to correct or ameliorate a drift in the first characteristic signal in view of the second characteristic signal. It is an advantage of such device that particularly long measurement times without signficant influence of drift can be achieved, thus facilitating or even enabling to detect target substances at low concentrations at an investigation region or against a noisy medium background.
- the sensing device is adapted to cooperate with an analyzing device for detecting the target substance, wherein the analyzing device comprises::
- a light source to direct incident light to the analysis region such that the incident light is directed to the investigation region and the reference region of the sensing surface and such that the incident light is reflected under total internal reflection conditions at the investigation region and the reference region, thereby generating reflected light, when the sensing device is accommodated in the analysis region,
- a detector for detecting the reflected light to yield a first characteristic signal depending on the reflection at the investigation region and a second characteristic signal depending on the reflection at the reference region
- an analyzer device for detecting a target substance adapted to cooperate with a sensing device for detecting the target substance, wherein the sensing device comprises:
- the analyzing device comprises: - an analysis region for accommodating the sensing device,
- a light source to direct incident light to the analysis region such that the incident light is directed to the investigation region and the reference region of the sensing surface and such that the incident light is reflected under total internal reflection conditions at the investigation region and the reference region, thereby generating reflected light, when the sensing device is accommodated in the analysis region,
- a detector for detecting the reflected light to yield a first characteristic signal depending on the reflection at the investigation region and a second characteristic signal depending on the reflection at the reference region
- Such an analyzer device is advantageously adapted to a sensing device, particularly a cartridge, of the present invention and makes use of the reference region for calibrating a characteristic signal obtained from the investigation region.
- the characteristic signal preferably is a light intensity signal.
- the sensing device preferably is a biosensor.
- the calibrator is preferably adapted to correct or ameliorate a drift in the first characteristic signal of light emanating from an investigation region of the sensing device, particularly a cartridge, in view of the second characteristic signal of light emanating from a reference region of the sensing device .
- the analyzer device allows to reduce the measurement error for light intensity measurements at the investigation region. It is thus possible to analyze light reflected from the investigation region for a long period of time without significantly tainting the first characteristic signal obtained from said investigation region by erroneous light intensity drifts. Such analyzing device thus facilitates detection of substances at an investigation region at the very low concentrations which typically require a long irradiation time of the investigation region.
- a sensing method for detecting a target substance in an investigation region comprising the steps of
- the sensing method can further comprise the steps of: a) irradiating an investigation region of a sensing device of the invention with light and obtaining a first characteristic signal of totally internally reflected light therefrom, b) before, during or after step a) irradiating a reference region of the sensing device of the present invention with light and obtaining a second characteristic signal of totally internally reflected light therefrom, c) calibrating the first characteristic signal obtained in step a) by the second characteristic signal obtained in step b).
- the sensing method allows to detect the presence of a target substance at an investigation region, and also allows to determine another property of the investigation region, e.g. to determine a temperature by analysing the influence of a thermochromic substance on the intensity of light reflected under total internal reflection conditions at the investigation region.
- steps a) and c), steps b) and c) or steps a), b) and c) are repeated. Such repetition can be performed for one investigation region to allow a time- resolved analysis of target substance binding at the investigation region.
- the steps can also be repeated by applying them sequentially to different investigation regions, allowing to determine the amount of target substance(s) bound at different investigation regions.
- a sensing method comprising i) providing an analyzing device of the present invention, ii) providing a sensing device, preferably a cartridge, of the present invention at an analysis region of said analyzing device , iii) binding a target substance at an investigation region of said sensing device, iv) performing the measuring method steps a), b) and c) as described above.
- Such method exploits the advantages of the present invention and particularly allows analysis of a target substance at a very low concentration at an investigation region as described above.
- sensing device of claim 1 the analyzer device of claim 12 and the sensing method of claim 14 have similar and/or identical preferred embodiments as defined in the dependent claims. It shall further be understood that a preferred embodiment of the invention can also be any combination of the dependent claims with the respective independent claim.
- Fig. 1 shows schematically and exemplarily a side view of an embodiment of a sensing device
- Fig. 2 shows schematically and exemplarily a graph of a signal drift of an uncorrected detection signal
- Fig. 3 shows schematically and exemplarily a side view of light reflected under total internal reflection conditions
- Fig. 5 shows schematically and exemplarily a side view of the embodiment of a sensing device of figure 4,
- Fig. 6 shows schematically and exemplarily a top view of an embodiment of a sensing device
- Fig. 7 shows schematically and exemplarily a side view of the embodiment of a sensing device of figure 6
- Fig. 8 shows schematically and exemplarily a graph of a detection signal and a reference signal
- Fig. 10 shows schematically and exemplarily a top view of the embodiment of a sensing device of figure 9,
- Fig. 11 shows schematically and exemplarily a side view of an embodiment of a sensing device
- Fig. 12 shows schematically and exemplarily a side view of an embodiment of an analyzing device
- Fig. 13 shows schematically and exemplarily magnetic particles bound to an investigation region of a sensing device
- Fig. 14 shows schematically and exemplarily a method of the present invention.
- Fig. 1 shows schematically and exemplarily a side view of an embodiment of a sensing device 100 of the present invention.
- the device 100 comprises a carrier 110 of a material transparent for an incident light beam Ll .
- the carrier 110 has a sensing surface 112.
- a fluid delimiter 101 is positioned such as to allow a medium 4 to be added to the sensing surface 112.
- An area of the sensing surface 112 is covered by a reference element 121 to shield a reference region 120 of the sensing surface 112 from any target substance (2) in the medium 4.
- the investigation region 113 can comprise a binder 114 for direct or indirect binding of the target substance 2.
- Both the reference region 120 and the investigation region 113 can be illuminated by incident light (shown as incident light beam Ll).
- incident light beam Ll is reflected under total internal reflection conditions at the reference region 120 to produce outgoing light L2.
- the reference element 121 has such refractive index and such dimensions that an evanescent field elicited by total internal reflection at the reference region 120 remains unaffected by the presence or absence of the target substance 2.
- the intensity of the outgoing light beam L2 of the reference region 120 does not depend on the presence or absence of the target substance 2 in the medium 4, and is preferably also independent of the presence or absence of other substances of the medium 4.
- the outgoing light beam L2 of the reference region 120 can then serve as a standard light beam, and its intensity can be used as a (second) characteristic signal 220 at a detector 18.
- a small drop of UV-curable, low refractive index acrylate (2,2,3, 3,4,4, 5,5-Octafluoro-hexanediol-l,6-dimethacrylate) can be applied on a sensing surface 112 of a polystyrene cartridge. After curing under nitrogen conditions, a reference element 121 with n 2 ⁇ 1.42 at a reference region 120 can thus be obtained.
- a reference element 121 it was found that when an adjacent investigation region 113 of the sensing surface 112 was blackened with a black marker pen, the intensity of light reflected (L2) under total internal reflection conditions (angle of incidence B 1 : 70°) decreased markedly at the investigation region 113. However, even though the reference element 121 was also covered by ink of the black marker pen, the intensity of light reflected under total internal reflection conditions (angle of incidence Q 1 : 70°) did not decrease markedly at the reference region 120, as shown in figure 8.
- the refractive index n 2 of the cured acrylate is still rather high.
- the incoming beam was not perfectly parallel, such that part of the incident light had a smaller angle than 70°. This means that a small portion of the incoming light beam penetrated the reference element 121 and was thus subjected to the influence of the black marker ink, reducing the intensity of reflected light.
- Fig. 2 shows schematically and exemplarily a graph of a signal drift of an uncorrected detection signal.
- the x-axis refers to measurement time t given in minutes.
- the y-axis refers to the variation (given as a percentage) of a first characteristic signal 213 of light of an investigation region 113 of a sensing device 100 of the type of figure 1. No substance was added to the investigation region 113 during the measurement of figure 2.
- the graph shows that after 10 minutes of measuring time, the first characteristic signal 213 had drifted by 0.1%. Since this drift was not caused by the addition of a substance to the investigation region 113, the drift must be considered a measurement error.
- drift is considered to result from arbitrary variations in the intensity of a light source 11 , e.g. a LED, and the sensitivity of a sensor, e.g. a CMOS sensor, particularly due to temperature changes in the device.
- a light source 11 e.g. a LED
- a sensor e.g. a CMOS sensor
- Such measurement errors cannot be tolerated in analytical tasks requiring high detection sensitivity and precision, e.g. the determination of Troponin-I concentration in a medium 4 like blood.
- Such analytical tasks can require a detection limit of less than 1 pM which, in turn, require long measuring times with very low signal drift.
- the device of figure 1 now allows to reliably accomplish these analytical tasks.
- Fig. 3 shows schematically and exemplarily a side view of light reflected under total internal reflection conditions and illustrates the principle of total internal reflection.
- An incident light beam Ll travels through a medium 4 with a first refractive index ni.
- the light beam Ll reaches a surface to another medium 4 with a second refractive index n 2 .
- a material useful for manufacture of biosensor cartridges is polystyrene.
- ni is 1.55.
- the second medium 4 should be chosen such that n 2 ⁇ 1.45. ⁇ c ⁇ t would then be 69.1°, such that total internal reflection would occur at the surface between both media.
- n 2 some margin is included for n 2 , as there is frequently some angular distribution in the angle of incidence, e.g. as a LED might not produce perfectly parallel beams.
- n 2 should be chosen to be less than 1.419.
- Figures 4 and 5 schematically and exemplarily show a top and side view of a sensing device 100, respectively.
- the sensing device 100 has a cartridge 110 of a first refractive index ni.
- the cartridge 110 has a sensing surface 112.
- On the sensing surface 112, a reference element 121 is positioned to shield a reference area 120 of the sensing surface 112. Adjacent to the reference area 120 is an investigation region 113 of the sensing surface 112.
- the investigation region 113 does not comprise a reference element 121. In figure 5, the investigation region 113 is obscured by the reference element 121. Both the investigation region 113 and the reference region 120 can be illuminated by a beam of incident light Ll of a LED light source 11.
- the light source 11 can also be a laser diode or Super Luminescent Diode (SLED) or another light source 11.
- the incident light beam Ll is reflected under total internal reflection conditions at the reference region 120 and, in the absence of an influencing target substance (2), also at the investigation region 113 to form a light beam L2.
- the light beam L2 is detected at a detector 18 to determine the intensity of light reflected at the reference region 120 and investigation region 113, respectively.
- a calibrator 20 compares the outgoing light L2 of both the reference region 120 and the investigation region 113 as detailed in the discussion of figure 1, and as further detailed in the discussion of figure 12.
- Figures 6 and 7 show schematically and exemplarily a top view and side view, respectively, of an embodiment of a sensing device 100.
- the sensing device 100 comprises a carrier 110.
- the carrier 110 has a sensing surface 112.
- a fluid delimiter 101 is positioned such as to allow a preferably liquid medium 4 to be added to the sensing surface 112.
- the fluid delimiter 101 has openings forming a fluid channel system comprising a fluid reception opening 112, a channel leading to a measurement chamber, a channel leading away from the reference chamber, and a vent opening 119.
- the measurement chamber comprises an investigation region 113 of the sensing surface 112. Adjacent to the measurement chamber are two reference regions 120, each comprising a solid reference element to allow total internal reflection of incident light in the carrier 110.
- the investigation region 113 and one or both of the reference regions 120 are irradiated by an incident light beam (not shown) in the manner depicted in figure 1.
- the light is reflected under total internal reflection conditions at the reference region(s) 120 and/or the investigation region 113.
- a fluid medium 4 preferably a liquid, is added to the sensing device 100 via fluid reception opening 120. Air contained in the measurement chamber and channel leading thereto is expelled via vent opening 119.
- the medium 4 is channeled to the measurement chamber.
- a target substance 2 can influence the intensity of light reflected at the investigation region 113 of the sensing surface 112 under total internal reflection conditions.
- the intensities of light reflected under total internal reflection conditions at the investigation region 113 and the reference region(s) 120 are detected by a detector 18.
- a calibrator 20 compares the outgoing light of both the reference region(s) 120 and the investigation region 113 in the manner described for figure 1.
- Figure 8 shows schematically and exemplarily a graph of a detection signal and a reference signal of a device of figures 6 and 7.
- a normalized signal intensity S n is graphed.
- the characteristic signal 220 obtained from light reflected at a reference region 120 of the sensing surface 112 shows some drift in signal intensity. This detected drift in signal intensity can be used to correct and calibrate the characteristic signal 213 obtained from light reflected at the investigation region 113 of the sensing surface 112.
- Figures 9 and 10 show schematically and exemplarily a side view and top view, respectively, of a further embodiment of a sensing device 100.
- the device 100 comprises a carrier 110 having a sensing surface 112 thereon.
- the sensing surface 112 comprises a series of recesses to form a series of lowered reference regions 120 therein.
- the recesses may be formed by surface patterning, e.g. by using focus ion beam milling or pulsed laser ablation. Adjacent to the reference areas 120 is an investigation region 113 as described with regards to figure 1.
- a liquid medium 4 passes over the sensing surface 112 of the carrier 110.
- a droplet of medium 4 is shown in figures 9 and 10.
- the medium 4 cannot enter the recesses at the reference region 120 due to its surface tension, entrapping air between the medium droplet 4 and the reference region 120.
- the air then functions as reference element 121 as described regarding figure 1.
- FIG. 11 shows schematically and exemplarily a side view of a further embodiment of a sensing device 100.
- the device comprises a carrier 110 having a sensing surface 112 formed thereon.
- the sensing surface 112 comprises an investigation region 113 as described for figure 1, and a reference region 120.
- the sensing surface 112 of the reference region 120 is tilted relative to the sensing surface 112 of the investigation region 113 by an angle of ⁇ .
- Fig. 12 shows schematically and exemplarily a side view of an embodiment of an analyzer device 10.
- a sensing device 100 has been inserted in the analyzer device 10.
- the sensing device 100 comprises, in this embodiment, a carrier 110 having a sensing surface 112 thereon. Further, a fluid delimiter 101 is positioned on the carrier 110.
- a top fluidic part 14 completes a measuring chamber between the sensing surface 112, the fluid delimiter 101 and the top fluidic part 14.
- the analyzer device 10 further comprises a magnetic element 13 which provides a magnetic field for forcing magnetic particles 2 onto the sensing surface 112 of the sensing device.
- the magnetic particles 2 are detected by, in this embodiment, illuminating the sensing surface 112 with a light beam Ll generated by a light source 11.
- the light source 11 is, for example, a laser device, SLED or a LED.
- Light L2 reflected from the sensing surface 112 is detected by a detector 18.
- the detector 18 is, for example, a photodetector or a two-dimensional camera.
- Optical elements can be arranged in the light beams Ll and L2 for generating parallel light beams Ll and L2, respectively. Such optical elements are preferably lenses.
- the carrier 110 is accommodated in an analysis region.
- both incident light Ll can be directed at the reference region 120 and the investigation region 113 such that light reflected under total internal reflection conditions can be detected by the detector 18.
- the magnetic field created by magnetic element 13 can force magnetic particles 2 onto the sensing surface 112.
- the sensing surface 112 further comprises a reference region 120 and an investigation region 113; both are not shown. Incident light Ll arriving at the reference region 120 and investigation region 113 of the sensing surface 112 is reflected under total internal reflection conditions to become a light beam L2. However, at the investigation region 113 such internal reflection may be under frustrated total internal reflection conditions.
- the detector 18 yields two characteristic signals 213, 220.
- the intensity of light reflected under total internal reflection conditions at the investigation region 113 is yielded as first characteristic signal 213, the intensity of light reflected under total internal reflection conditions at the reference region 120 is yielded as second characteristic signal 220.
- a calibrator 20 then compares the first characteristic signal 213 and the second characteristic signal 220 to produce a corrected result. The result of the comparison is displayed by a display device 21.
- Fig. 13 shows schematically and exemplarily magnetic beads bound to an investigation region 113 of a sensing device.
- An investigation region 113 of a sensing surface 112 is coated with antibodies 114.
- the antibodies 114 can bind an analyte 2'.
- magnetic beads 2 are coated with antibodies to bind analyte 2'.
- analyte 2' is sandwiched between antibodies 114 and magnetic beads 2, thus effectively coupling the magnetic beads 2 to the investigation region 113.
- the magnetic beads 2 can then influence the light reflected at the investigation region 113 as described with respect e.g. to figure 12.
- the magnetic beads are preferably particles having at least one dimension ranging between 3 nm and 10000 nm, preferably between 10 nm and 3000 nm, and more preferably between 200 nm and 1000 nm.
- Fig. 14 shows schematically and exemplarily a method of the present invention.
- a sensing surface 112 with an investigation region 113 and reference region 120 is provided.
- a reference element 121 is located at the reference region 120 to shield the reference region 120 such that light reflected at the reference region 120 under total internal reflection conditions remains unaffected by the presence or absence of a target substance 2 or coloured particulates of a medium 4 comprising the target substance 2 and, where applicable, an analyte.
- Providing such reference element 121 at the reference region 120 may in some embodiments be performed as an independent method step.
- the investigation region 113 and the reference region 120 are illuminated under total internal reflection conditions in step 402.
- a detector 18 detects light reflected from the investigation region 113 (step 403) and the reference region 120 (step 404) under total internal reflection conditions. Steps 403 and 404 can be performed simultaneously, e.g. using a CCD detector.
- a first characteristic signal 213 is obtained from the detector 18 depending on the light reflected from the investigation region 113.
- a second characteristic signal 220 is obtained from the detector 18 depending on the light reflected from the reference region 120.
- the first 213 and second 220 characteristic signals are read by a calibrator 20.
- the calibrator 20 calibrates the first characteristic signal 213 in view of the second characteristic signal 220.
- a calibrated first characteristic signal 213 is output by the calibrator 20.
- the medium 4 was preferably blood.
- the fluid can be any other fluid, in particular any other body fluid, for example saliva or urine.
- the preferred application of the sensing and analyzer device 10 is in the field of point of care diagnostics and detection of drugs or the detection of the occurence of a heart attack, and in particular based on finger prick blood samples and saliva samples.
- the device, methods and systems of this invention are suited for sensor multiplexing (i.e. the parallel use of different sensors and sensor surfaces), label multiplexing (i.e. the parallel use of different types of labels as target substances 2) and chamber multiplexing (i.e. the parallel use of different reaction chambers).
- the devices and methods described in the present invention can be used as rapid, robust, and easy to use point-of-care biosensors for small sample volumes.
- the carrier can be a disposable item to be used with a compact reader, containing one or more magnetic field generating means and one or more detection means.
- the devices and methods of the present invention can be used in automated high-throughput testing.
- the carrier is e.g. a well plate or cuvette type carrier, fitting into an automated instrument.
- the device uses evanescent field techniques for determining the amount of magnetic particles on the investigation region 113 of the sensing surface 112. In other embodiments, other substances may be detected.
- larger moieties can be detected, e.g. cells, viruses, or fractions of cells or viruses, tissue extracts and so on.
- the embodiments have been described with reference to a sandwich assay as described in figure 13.
- other assay types can be employed, e.g. binding/unbinding assay, sandwich assay, competition assay, displacement assay, enzymatic assay, etc.
- the target substance 2 may be directly analyzed by its presence on the investigation region 113 without use of magnetic beads. This is particularly preferred for target substances 2 with a high ability to absorb or scatter light of an evanescent field on the investigation region 113.
- the target substance 2 can be further processed prior to detection. An example of further processing is that further materials are added or that the (bio)chemical or physical properties of the target substance 2 are modified to facilitate detection.
- the target substance 2 may for example be reacted with a reactant to alter its ability to absorb or scatter light of an evanescent field on the investigation region 113.
- the target substance 2 may be covalently linked to a light absorbing or scattering substance to influence an evanescent field in the investigation region 113.
- the detection can occur with or without scanning of the sensor element with respect to the sensing surface 112, in particular the reference region 120 and/or the investigation region 113.
- the reference element 121 has been described as being an element transparent to incident light but chosen to effect reflection under total internal reflection conditions.
- the reference element 121 may also be a true mirror 121, preferably a metal layer, to reflect incident light. This abolishes the need to select such materials for the manufacture of the reference element 121 that have a sufficient refractive index in view of the refractive index of the sensing surface material and the incident light.
- a single unit or device may fulfill the functions of several items recited in the claims.
- the mere fact that certain measures are recited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage.
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- Physics & Mathematics (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
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Abstract
Description
Claims
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
BRPI1006390A BRPI1006390A2 (en) | 2009-02-18 | 2010-02-08 | detection device for detecting a target substance in a research region, analyzer device for detecting a target substance and detection method for detecting a substance in a research region |
US13/201,927 US9804089B2 (en) | 2009-02-18 | 2010-02-08 | Sensing device for detecting a target substance |
CN201080008124.4A CN102317758B (en) | 2009-02-18 | 2010-02-08 | Sensing device for detecting objects matter |
EP10704203A EP2399116A1 (en) | 2009-02-18 | 2010-02-08 | Sensing device for detecting a target substance |
RU2011138146/28A RU2519505C2 (en) | 2009-02-18 | 2010-02-08 | Sensor device for target substance identification |
JP2011549712A JP5701778B2 (en) | 2009-02-18 | 2010-02-08 | Detection device for detecting target substances |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP09153105.3 | 2009-02-18 | ||
EP09153105A EP2221603A1 (en) | 2009-02-18 | 2009-02-18 | Sensing device for detecting a target substance |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2010095071A1 true WO2010095071A1 (en) | 2010-08-26 |
Family
ID=40848456
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/IB2010/050553 WO2010095071A1 (en) | 2009-02-18 | 2010-02-08 | Sensing device for detecting a target substance |
Country Status (7)
Country | Link |
---|---|
US (1) | US9804089B2 (en) |
EP (2) | EP2221603A1 (en) |
JP (1) | JP5701778B2 (en) |
CN (1) | CN102317758B (en) |
BR (1) | BRPI1006390A2 (en) |
RU (1) | RU2519505C2 (en) |
WO (1) | WO2010095071A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2014525572A (en) * | 2011-08-31 | 2014-09-29 | フラウンホッファー−ゲゼルシャフト ツァー フェーデルング デア アンゲバンテン フォルシュング エー ファー | Method and apparatus for determining a substance by using terahertz radiation |
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EP2567216A2 (en) * | 2010-05-03 | 2013-03-13 | Koninklijke Philips Electronics N.V. | Sensing device for detecting a substance in a fluid |
EP2527814A1 (en) * | 2011-04-27 | 2012-11-28 | Koninklijke Philips Electronics N.V. | Sensor system with an exchangeable cartridge and a reader |
JP5772238B2 (en) * | 2011-06-01 | 2015-09-02 | ウシオ電機株式会社 | Microchip for surface plasmon resonance measurement and surface plasmon resonance measurement apparatus |
EP2726852B1 (en) | 2011-06-30 | 2020-11-25 | Koninklijke Philips N.V. | Multiple examinations of a sample |
WO2013048328A1 (en) * | 2011-09-30 | 2013-04-04 | General Electric Company | Systems and methods for self-referenced detection and imaging of sample arrays |
CN105745526B (en) * | 2014-01-09 | 2018-11-30 | 夏普株式会社 | Light intensity sensing device and detection method |
CN108449968B (en) * | 2015-06-12 | 2021-08-17 | 莱克瑞科学有限责任公司 | Handheld field portable surface plasma resonance device and application thereof in detection of chemical and biological reagents |
DE102016101554A1 (en) * | 2016-01-28 | 2017-08-03 | Vorwerk & Co. Interholding Gmbh | Cleaning device, in particular wet mop |
CN113588602B (en) * | 2021-07-08 | 2024-06-04 | 合肥新唯基因科技有限公司 | Rapid parathyroid hormone detection method |
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- 2010-02-08 CN CN201080008124.4A patent/CN102317758B/en not_active Expired - Fee Related
- 2010-02-08 EP EP10704203A patent/EP2399116A1/en not_active Withdrawn
- 2010-02-08 JP JP2011549712A patent/JP5701778B2/en not_active Expired - Fee Related
- 2010-02-08 US US13/201,927 patent/US9804089B2/en active Active
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Also Published As
Publication number | Publication date |
---|---|
JP5701778B2 (en) | 2015-04-15 |
JP2012518169A (en) | 2012-08-09 |
CN102317758A (en) | 2012-01-11 |
EP2399116A1 (en) | 2011-12-28 |
EP2221603A1 (en) | 2010-08-25 |
RU2519505C2 (en) | 2014-06-10 |
BRPI1006390A2 (en) | 2019-09-24 |
RU2011138146A (en) | 2013-03-27 |
US9804089B2 (en) | 2017-10-31 |
CN102317758B (en) | 2016-06-01 |
US20120040475A1 (en) | 2012-02-16 |
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