JP2016080497A - Detection method of detection object - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a detection method of a detection object capable of excellently detecting the detection object even when the detection object is not colored.SOLUTION: A detection method of a detection object detects a detection object included in a dispersion dispersed with the detection object, and includes: an oscillation part 13 for generating a terahertz wave having a wavelength equal to or more than the maximum particle diameter of the detection object and irradiating the terahertz wave to the dispersion; a specimen supply part 14 for holding the dispersion so as to be irradiated with the terahertz wave generated by the oscillation part; and a detection part 15 for receiving a scattered wave of the terahertz wave irradiated to the detection object and detecting the detection object. The specimen supply part is composed of a transmission material that transmits the terahertz wave, includes a specimen supply part main body having an incident surface having a normal line along the advancing direction of the terahertz wave, and provided with a flow path for irradiating the terahertz wave to the dispersion, while moving the dispersion inside the specimen supply main body.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a method for detecting a detection object. More specifically, the present invention relates to a detection object detection method that can be suitably used for testing reagents, foods, etc., and obtaining information for assisting diagnosis of animal diseases.

  Flow cytometry is one of the methods for counting, sorting, or detecting detection objects contained in a dispersion. For example, detection of foreign substances in reagents and foods, detection of markers contained in biological samples It is used for etc. In the flow cytometry, generally, a detection target is detected by staining the detection target with a fluorescent reagent or the like and measuring fluorescence or the like derived from the fluorescent reagent. For example, if bovine develops mastitis, the number of somatic cells in the raw milk increases, so bovine mastitis testing involves staining somatic cells with a fluorescently labeled antibody and the fluorescence of the stained somatic cells. Is detected by flow cytometry (see, for example, Patent Document 1).

  However, when flow cytometry is used, it is necessary to stain the detection target with a fluorescent reagent or the like when detecting the detection target included in the sample. There are things that cannot be done.

JP 2012-126657 A

  The present invention has been made in view of the above-described prior art, and it is an object of the present invention to provide a means that can detect a detection target satisfactorily without staining the detection target contained in a dispersion. And

The present invention
(1) A detection object detection method for detecting a detection object contained in a dispersion in which the detection object is dispersed, wherein a terahertz wave having a wavelength equal to or greater than a maximum particle diameter of the detection object is dispersed. And detecting a detection target by receiving a scattered wave of the terahertz wave applied to the detection target, and (2) the detection method according to (1) A detection object detection device used in the detection object detection method of
An oscillating unit that generates a terahertz wave having a wavelength equal to or greater than a maximum particle diameter of the detection target, and irradiates the dispersion with the terahertz wave;
A sample supply unit that holds the dispersion so that the terahertz wave generated by the oscillation unit is irradiated;
A detection unit that receives the scattered wave of the terahertz wave irradiated to the detection target and detects the detection target;
The sample supply unit is made of a transmission material that transmits the terahertz wave, and has a sample supply unit main body having an incident surface having a normal line along the traveling direction of the terahertz wave.
The present invention relates to a detection target object detection apparatus, wherein a flow path for irradiating the dispersion with terahertz waves while moving the dispersion in the sample supply unit main body is provided.

  According to the present invention, there is an excellent effect that the detection object can be detected well without staining the detection object contained in the dispersion.

It is a schematic explanatory drawing which shows an example of the whole structure of the detection apparatus used for the detection method of the detection target object which concerns on one Embodiment of this invention. It is a schematic explanatory drawing which shows an example of a principal part structure of the detection apparatus used for the detection method of the detection target which concerns on one Embodiment of this invention. It is a schematic explanatory drawing which shows the modification of the principal part structure of the detection apparatus used for the detection method of the detection target object concerning one Embodiment of this invention. It is a schematic explanatory drawing which shows the whole structure of the detection apparatus used in Example 1. FIG. FIG. 3 is a schematic explanatory diagram illustrating a configuration of a sample supply unit of the detection device used in Example 1. In Example 1, a graph showing the results of examining the relationship between the wavelength and the S ₄₀₀ / S _30. It is a schematic explanatory drawing which shows the whole structure of the detection apparatus used in Example 2. In Example 2, it is a graph which shows an example of the result of having investigated the time-dependent change of the voltage value. In Example 2, it is a graph which shows the result of having investigated the relationship between the kind of test sample, and scattering intensity / transmission intensity. It is a schematic explanatory drawing which shows the whole structure of the detection apparatus used in Example 3 and 4. In Example 3, it is a graph which shows the result of having investigated the relationship between the kind of test sample, and scattering intensity / transmission intensity. In Example 4, it is a graph which shows the result of having investigated the relationship between the kind of test sample, and scattering intensity / transmission intensity.

  A detection target object detection method according to an embodiment of the present invention is a detection target detection method for detecting a detection target object included in a dispersion in which the detection target object is dispersed. The present invention is characterized in that a terahertz wave having a wavelength equal to or larger than the maximum particle diameter is irradiated onto a dispersion, and the detection target is detected by receiving a scattered wave of the terahertz wave irradiated on the detection target.

  The detection target object detection method according to the present embodiment irradiates a dispersion in which the detection target object is dispersed with a terahertz wave having a wavelength equal to or greater than the maximum diameter of the detection target object, and a scattered wave emitted from the detection target object. Therefore, the detection target contained in the dispersion can be detected satisfactorily.

  The maximum diameter of the detection target is 10 μm to 1 mm or more from the viewpoint of improving the intensity of the scattered wave when the terahertz wave is irradiated. Since the maximum diameter of the detection object varies depending on the wavelength of the terahertz wave used, etc., it is preferable that the maximum diameter is appropriately determined according to the wavelength of the terahertz wave used. The maximum diameter of the detection target can be obtained by observing under a microscope.

  Examples of the detection target include cells such as epithelial cells and somatic cells such as leukocytes, but the present invention is not limited to such examples.

  Examples of the dispersion include a dispersion containing a detection target and the detection target dispersed in a dispersion medium (for example, milk containing somatic cells as the detection target). The invention is not limited to such examples.

  Further, the dispersion may further contain a non-detection object smaller than the detection object. In the present specification, “smaller than the detection object” means smaller than the maximum diameter of the detection object. Examples of the non-detection target include proteins and lipids, but the present invention is not limited to such examples.

  When the dispersion includes a solvent as a dispersion medium, examples of the solvent include water; alcohols such as methanol and ethanol. However, the present invention is not limited to such examples.

  The wavelength of the terahertz wave applied to the dispersion is a wavelength equal to or greater than the maximum diameter of the detection target. Since the wavelength of the terahertz wave differs depending on the type of detection object, the maximum diameter of the detection object, the type of dispersion, etc., it cannot be determined unconditionally, so the type of detection object, the detection object It is preferable to determine appropriately according to the maximum diameter, the type of dispersion, and the like. The wavelength of the terahertz wave is usually 30 μm to 3 mm. For example, when the dispersion is milk and the detection target is a cell, the size of the cell is usually 10 to 30 μm, and therefore the wavelength of the terahertz wave is preferably 30 to 100 μm. .

  The size of the wavelength of the terahertz wave with respect to the maximum diameter of the detection target is one or more times from the viewpoint of improving the scattering intensity and detecting the detection target with higher sensitivity. The upper limit of the wavelength of the terahertz wave with respect to the maximum diameter of the detection target can be appropriately determined within a range where the scattering intensity can be improved and the detection target can be detected with higher sensitivity.

  Since the intensity of the terahertz wave irradiated to the dispersion varies depending on the type of the detection target and the like, it cannot be determined unconditionally. Therefore, it is preferable to appropriately determine the intensity according to the type of the detection target.

  From the viewpoint of detecting the detection target contained in the dispersion with higher sensitivity, the dispersion moves in the flow path while introducing and moving the dispersion in the flow path made of a transmission material that transmits the terahertz wave. It is preferable that the terahertz wave is irradiated. As the flow path, for example, a flow path included in a detection device described later can be used. Since the flow rate of the dispersion in the flow path varies depending on the type of the detection target object and the like, it cannot be generally determined. Therefore, it is preferable to appropriately determine the flow rate of the dispersion according to the type of the detection target object.

  Examples of the transmissive material include silicon; (meth) acrylic resins such as polymethyl methacrylate; polyester resins such as polyethylene terephthalate and polybutylene terephthalate; polyamide resins such as nylon (registered trademark of DuPont); polyethylene and polypropylene However, the present invention is not limited to such examples. In this specification, the (meth) acrylic resin refers to an acrylic resin or a methacrylic resin.

  Examples of the shape of the cross section perpendicular to the flow direction of the flow path include a circular shape and a quadrangular shape, but the present invention is not limited only to such illustration. From the viewpoint of suppressing a decrease in detection sensitivity due to refraction of the terahertz wave, the shape of the cross section perpendicular to the flow direction of the flow path is preferably a quadrangular shape. When the cross-sectional shape perpendicular to the flow direction of the flow path is a quadrangle, from the viewpoint of suppressing a decrease in detection sensitivity due to refraction of the terahertz wave, the incident surface of the terahertz wave to the flow path is the traveling direction of the terahertz wave Are preferably provided so as to have a normal line.

  When the shape of the cross section perpendicular to the flow direction of the flow path is circular, the inner diameter of the flow path varies depending on the type of the detection target, and therefore cannot be determined unconditionally. It is preferable to determine appropriately according to the kind of the. Usually, the inner diameter of the flow path is preferably 500 to 1000 μm.

  Next, an example of a detection apparatus used in the detection method of the detection target object according to the embodiment of the present invention will be described with reference to the accompanying drawings. FIG. 1 is a schematic explanatory diagram showing an example of the entire configuration of a detection apparatus used in the detection target object detection method according to this embodiment of the present invention. In the following, a detection device using a terahertz wave generated using femtosecond pulsed laser light as excitation light will be described as an example, but the present invention is not limited to such an example. . A detection apparatus 1 shown in FIG. 1 generates a terahertz wave by receiving a light source 11 that emits femtosecond pulsed laser light, which is excitation light, and a femtosecond pulsed laser light emitted from the light source 11. An oscillating unit 13 that irradiates, a sample supply unit 14, a detection unit 15 that receives scattered waves, a control unit 16, an output unit 17, and a power source (not shown) are provided.

  The femtosecond pulsed laser light L1 emitted from the light source 11 is incident on the oscillation unit 13. The oscillation unit 13 generates a terahertz wave L3 when the pump light L2 is incident. The detection unit 15 receives the scattered wave L4 generated by irradiating the sample moving in the flow path of the sample supply unit 14 with the terahertz wave L3, whereby a voltage having a magnitude corresponding to the intensity of the scattered wave L4 is generated. Occur. In the detection apparatus 1 shown in FIG. 1, a frequency spectrum can be obtained by Fourier transforming the electric field amplitude waveform in the time domain.

  Examples of the light source 11 include a solid-state laser such as a titanium sapphire laser; a fiber laser such as an erbium-doped fiber laser; and a semiconductor laser such as a quantum cascade laser. However, the present invention is not limited to such an example. Absent.

  Since the wavelength of the femtosecond pulsed laser light emitted from the light source 11 differs depending on the type of the oscillating unit 13 and the like, it cannot be determined unconditionally. Therefore, it can be appropriately determined according to the type of the oscillating unit 13 and the like. preferable.

  In the present invention, a coherent light source such as a light emitting diode may be used as the light source 11 instead of a light source that emits femtosecond pulsed laser light. Since the wavelength of the excitation light that is discarded from the coherent light source varies depending on the type of the oscillating unit 13 and the like, it cannot be determined unconditionally.

  Examples of the oscillating unit 13 include a photoconductive antenna; a nonlinear optical crystal such as 4-dimethylamino-N-methyl-4-stilbazolium tosylate, but the present invention is limited only to such examples. is not. In the detection device 1 shown in FIG. 1, the femtosecond pulsed laser light L <b> 1 emitted from the light source 11 is incident on the oscillation unit 13. The oscillation unit 13 generates a terahertz wave L3 when the pump light L2 is incident.

  The sample supply unit 14 holds a dispersion that is a sample so that the terahertz wave is irradiated. The sample supply unit 14 is made of a transmissive material that transmits terahertz waves. The permeable material in the sample supply unit 14 is the same as the permeable material in the channel.

  The sample supply unit 14 can irradiate a terahertz wave to the dispersion moving in the flow channel while moving the dispersion from the viewpoint of detecting the detection target contained in the dispersion with higher sensitivity. It is preferable to have. An example of the sample supply unit 14 is a sample supply unit 14a shown in FIG. A sample supply unit 14a shown in FIG. 2 includes a block-shaped sample supply unit main body 101 made of the permeation material, a flow channel 102 for conveying a dispersion as a sample, and a sample connected to the upstream side of the flow channel 102. It has an introduction flow path 104, a sample discharge flow path 105 connected to the downstream side of the flow path 102, an incident surface 103 on which terahertz waves are incident, and an output surface 106 from which scattered waves are emitted. In the sample supply unit 14 a shown in FIG. 2, the incident surface 103 is provided so as to have a normal line in the traveling direction of the terahertz wave in the block-shaped sample supply unit main body 101. Further, a lens 103 a is formed on the incident surface 103 so that the terahertz wave is converged on the dispersion that moves in the flow path 102. As described above, the dispersion surface is provided so that the normal line of the incident surface 103 is along the traveling direction of the terahertz wave in the block-shaped sample supply unit main body 101 and moves in the flow channel 102 to the incident surface 103. Since the lens 103 a is formed so as to converge the terahertz wave, the refraction of the terahertz wave can be suppressed, the terahertz wave can be reliably incident on the sample supply unit 14 a, and the dispersion moves in the channel 102. The terahertz wave can be easily focused on. Therefore, according to the detection apparatus having the sample supply unit 14a shown in FIG. 2, it is possible to irradiate the detection target S with the terahertz wave in a better state, and to generate the scattered wave more reliably. The detection object S can be detected with higher sensitivity. The exit surface 106 can be provided so as to be parallel to the entrance surface 103. As described above, when the exit surface 106 is provided so as to be parallel to the entrance surface 103, the scattered wave emitted from the detection target can be easily reached by the detection unit 15. Further, from the viewpoint of suppressing a decrease in detection sensitivity due to terahertz wave refraction, the sample supply unit 14 is formed in a flow path (in the drawing, the sample supply unit main body is It is preferable to have an incident surface having a normal direction in the traveling direction of the terahertz wave and an output surface provided in parallel with the incident surface.

  The detection unit 15 receives the scattered wave L4 emitted from the detection target S included in the dispersion. The detection unit 15 receives the scattered wave L4 to cause a change in current or a change in voltage. According to the detection apparatus 1 shown in FIG. 1, the presence / absence of the detection target and the amount of the detection target can be evaluated using the change in current or voltage. Examples of the detection unit 15 include a photoconductive antenna, but the present invention is not limited to this example. In the detection device 1 shown in FIG. 1, a plurality of detection units 15a1 and 15a2 may be provided on the opposite side of the oscillation unit 13 with the flow channel 102 therebetween (see FIG. 2). The detection unit may be a detection unit 15b in which a plurality of detection units 15b1, 15b2, 15b3, and 15b4 are arranged adjacent to each other (see FIG. 3). In FIG. 3, the sample supply unit 14b has the same configuration as the sample supply unit 14a shown in FIG.

  The control unit 16 uses the intensity of the scattered wave detected by the detection unit 15 to generate information about the detection target contained in the dispersion and causes the output unit 17 to output the information. Examples of the control unit 16 include a computer, but the present invention is not limited to such an example. Examples of such information include the amount of a detection target, but the present invention is not limited to such examples.

  The output unit 17 outputs the information generated by the control unit 16. Examples of the output unit 17 include a monitor and a printer. However, the present invention is not limited to such examples.

  As described above, according to the detection target object detection method and detection apparatus according to an embodiment of the present invention, the detection target object can be detected satisfactorily without staining the detection target object included in the dispersion. can do. Therefore, the sample used for the detection method of the detection target object which concerns on one Embodiment of this invention can be used for another use. Therefore, the present invention is useful for examination of reagents, foods, etc., acquisition of information for assisting diagnosis of animal diseases and the like.

  Next, the present invention will be described in more detail based on examples, but the present invention is not limited to such examples. In the following, the maximum diameter is determined by observing under a microscope. The average particle diameter is measured by a laser diffraction / scattering method.

Example 1
(1) Sample preparation Maximum particle diameter of soda-lime glass: 30 μm beads [manufactured by Unitika Ltd., wavelength: 300 μm, refractive index: 1.9] and thousands of soda-lime glass maximum particles One bead having a diameter of 400 μm (manufactured by Unitika Co., Ltd., wavelength: refractive index at 300 μm: 1.9) was suspended in isobutanol to obtain a dispersion as a sample. In the following, beads having a particle diameter of 400 μm were used as detection objects, and beads having a particle diameter of 30 μm were used as non-detection objects.

(2) Construction of detection device The detection device 2 shown in FIG. 4 was constructed. 4 includes a light source 11 that emits femtosecond pulsed laser light, a beam splitter 12 that divides the femtosecond pulsed laser light, and an oscillation that receives the femtosecond pulsed laser light and generates a terahertz wave. A unit 13, a sample supply unit 14, a detection unit 15 that receives scattered waves, a control unit 16, an output unit 17, and a time delay unit 18 are provided. 4 includes a voltage source for applying a voltage to both the oscillation unit 13 and the detection unit 15 (not shown). In the detection apparatus 2 shown in FIG. 4, the femtosecond pulsed laser light L1 emitted from the light source 11 is split by the beam splitter 12 into pump light L2 and probe light L5. The oscillation unit 13 generates a terahertz wave L3 when the pump light L2 is incident. On the other hand, the time delay unit 18 appropriately adjusts the timing at which the probe light L6 that has passed through the time delay unit 18 reaches the detection unit 15 by changing the optical path length of the probe light L5. The detection unit 15 receives the scattered wave L4 generated by irradiating the sample moving in the flow path of the sample supply unit 14 with the terahertz wave L3 and receives the probe light L6, thereby increasing the intensity of the scattered wave L4. A voltage of a corresponding magnitude is generated. According to the detection device 2 shown in FIG. 4, a frequency spectrum can be obtained by Fourier transforming the electric field amplitude waveform in the time domain.

  In the detection apparatus 2 shown in FIG. 4, a femtosecond pulse laser light source (manufactured by Integral, trade name: FEMTOSOURCE integral PRO) as the light source 11, and a beam splitter (manufactured by Hamamatsu Photonics Co., Ltd.) as the beam splitter 12. ] A photoconductive antenna (manufactured by Hamamatsu Photonics Co., Ltd.) was used as the oscillation unit 13 and the detection unit 15.

  Further, the sample supply unit 14 c shown in FIG. 5 was used as the sample supply unit 14. The sample supply unit 14 shown in FIG. 5 is provided so as to have a normal direction in the traveling direction of the terahertz wave L3 and the sample supply unit main body 111, the channel 112 formed in the sample supply unit main body 111. The incident surface 113 and an exit surface 114 provided in parallel to the incident surface 113. Thin portions 113b and 114b are formed so as to have a thickness of 3 mm at portions of the entrance surface 113 and the exit surface 114 that overlap the optical path of the terahertz wave L3. By providing such thin portions 113b and 114b, it is possible to suppress the influence of the sample supply unit main body 111 on the terahertz wave. The inner diameter of the flow path 112 is 500 μm. A sample introduction channel 115 and a sample discharge channel 116 are connected to the channel 112. As the sample supply unit main body 111, a fine channel [manufactured by Pax, wavelength: refractive index at 300 μm: 1.5] was used.

(3) Receiving scattered waves While introducing and moving the sample into the flow path 112 of the detection device 2, the sample is irradiated with electromagnetic waves (terahertz waves) having a wavelength of 30 to 3000 μm and emitted from beads contained in the sample. The voltage based on the scattered wave that was generated was measured. Using the measured voltage value, the intensity of the scattered wave (hereinafter also referred to as “scattering intensity”) was obtained. Next, using the measured values of the scattering intensity of the beads having a particle diameter of 400 μm (hereinafter referred to as “S ₄₀₀ ”) and the scattering intensity of the beads having a particle diameter of 30 μm (hereinafter referred to as “S ₃₀ ”), S ₄₀₀ / to calculate the S _30. FIG. 6 shows the result of examining the relationship between the wavelength and S ₄₀₀ / S ₃₀ . Figure, A represents S ₄₀₀ and S ₃₀ S ₄₀₀ / S ₃₀ calculated from the respective measured values, B is the light scattering by the literature [ "small particles Harusuto (H.C. van de Hulst) et (Light Scattering By Small the particles) ", Dover Books on Physics (Dover Books on Physics), the scattering intensity and wavelength and the theoretical value of S ₄₀₀ / S ₃₀ obtained according to the relationship equation between the particle diameter according to published 1957) Indicates.

From the results shown in FIG. 6, it can be seen that when the wavelength of the irradiated electromagnetic wave is larger than the maximum diameter (400 μm) of the detection object, the difference between S ₄₀₀ and S ₃₀ becomes large. From these results, irradiate the dispersion containing the detection target and non-detection target with a terahertz wave having a wavelength larger than the maximum diameter of the detection target, and measure the scattered wave emitted from the detection target. This suggests that the detection target contained in the dispersion can be detected separately from the non-detection target. Moreover, it can be seen from the results shown in FIG. 6 that S ₄₀₀ / S ₃₀ calculated from the measured values of S ₄₀₀ and S ₃₀ substantially coincide with the theoretical value of S ₄₀₀ / S ₃₀ .

Example 2
(1) Preparation of sample A human-derived cell DLD-1 was suspended in a medium for DLD-1 so that its concentration would be 3 × 10 ⁶ cells / mL to obtain a dispersion as a sample.

(2) Construction of detection device The detection device shown in FIG. 7 was constructed. 7 includes an oscillation unit 51 that generates a terahertz wave having a wavelength larger than that of the human-derived cell DLD-1 (maximum diameter: 10 μm), a lens 52 that collects the terahertz wave, and a terahertz wave. A chopper 53 to be modulated, a sample supply unit 54, an integrating sphere 55, a scattered wave detection unit 56 that detects scattered waves, and a transmitted light detection unit 57 that detects transmitted light are provided. In the detection device 50 shown in FIG. 5, the terahertz wave L <b> 10 emitted from the oscillating unit 51 passes through the lens 52 and is collected on the sample placed in the sample supply unit 54. The terahertz wave L10 is synchronously detected by the chopper 53. The scattered wave L11 generated by irradiating the sample with the terahertz wave L10 is reflected inside the integrating sphere 55 and detected by the scattered wave detection unit 56. Further, the transmitted light L12 generated by irradiating the sample with the terahertz wave L10 passes through the integrating sphere 55 and is detected by the transmitted light detection unit 57. By receiving the scattered wave L11, the scattered wave detection unit 56 generates a voltage having a magnitude corresponding to the intensity of the scattered wave L11. In addition, the transmitted light detection unit 57 receives the transmitted light L12 and generates a voltage having a magnitude corresponding to the intensity of the transmitted light L12.

  In the detection device 50 shown in FIG. 4, the oscillation unit 51 includes an oscillation unit having a quantum cascade laser light source (oscillation wavelength: 4.57 μm) as a light source, and a transmission cell [optical path length: 100 μm, thickness as a sample supply unit 54]. The DLATAGS (deuterated L-alanine doped triglycine sulfate crystal) pyroelectric detector was used as the scattered wave detector 56 and the transmitted light detector 57.

(3) Measurement of scattered wave and transmitted light While introducing and moving the sample obtained in Example 2 (1) into the sample supply unit 54 of the detection device 50 obtained in Example 2 (2) Then, the sample in the sample supply unit 54 was irradiated with terahertz waves (wavelength 400 μm), and the voltage based on the scattered waves of the cells contained in the sample and the voltage based on the transmitted light were measured over time. An example of the result of examining the change in voltage value over time is shown in FIG. In the figure, A shows a change with time of the voltage value based on transmitted light, and B shows a change with time of the voltage value based on scattered waves.

  From the results shown in FIG. 9, when a scattered wave and transmitted light are emitted by irradiating a cell with a terahertz wave, the voltage value based on the transmitted light and the peak of the voltage value based on the scattered wave (A2 and B2). ), But you can see that. On the other hand, when the terahertz wave is not applied to the cell (A1 and B1), it can be seen that the voltage value based on the transmitted light and the voltage value based on the scattered wave hardly change.

  The same operation as described above was performed except that distilled water or a medium for DLD-1 was used instead of using the sample obtained in Example 2 (1), and the voltage based on the transmitted light and the voltage based on the scattered wave Was measured over time. Next, using the voltage value based on the transmitted light and the voltage value based on the scattered wave, the scattering intensity and the intensity of the transmitted light (hereinafter also referred to as “transmission intensity”) were obtained. Thereafter, the scattering intensity / transmission intensity was determined using the scattering intensity and the transmission intensity. FIG. 9 shows the results of examining the relationship between the type of test sample and the scattering intensity / transmission intensity. In the figure, 1 is the scattering intensity / transmission intensity when using the sample obtained in Example 2 (1), 2 is the scattering intensity / transmission intensity when using distilled water, and 3 is the medium for DLD-1. The scattering intensity / transmission intensity when using is shown.

  From the results shown in FIG. 9, the scattering intensity / transmission intensity when using the sample obtained in Example 2 (1) is the scattering intensity / transmission intensity when using distilled water and for DLD-1. It can be seen that it is significantly higher than the scattering intensity / transmission intensity when the medium is used. From the results shown in FIG. 9 and the results obtained in Example 1, the scattered wave emitted from the detection object by irradiating the terahertz wave having a wavelength larger than the maximum diameter of the detection object such as a cell, It can be seen that the detection target contained in the sample can be detected. Accordingly, the dispersion containing the detection target is irradiated with a terahertz wave having a wavelength larger than the maximum diameter of the detection target, and the scattered wave emitted from the detection target is measured, thereby detecting the detection target included in the dispersion. It is suggested that can be detected separately from non-detected objects.

Example 3
(1) Preparation of sample A human-derived cell DLD-1 (maximum diameter: 10 μm) was suspended in a medium for DLD-1 so that its concentration was 3 × 10 ⁶ cells / mL, and a dispersion as a sample was obtained. .

(2) Construction of detection device The detection device shown in FIG. 10 was constructed. A detection device 50 shown in FIG. 10 includes an oscillation unit 61 that generates a terahertz wave having a wavelength larger than that of the human-derived cell DLD-1 (maximum diameter: 10 μm), a lens 62, a chopper 63 that modulates the terahertz wave, An aperture 64 that narrows down the terahertz wave, mirrors 65a and 65b that change the optical path and adjust the optical axis, a sample supply unit 66, an integrating sphere 67, a scattered wave detection unit 68, and a transmitted light detection unit 69 are provided. Yes. In the detection device 60 shown in FIG. 10, the terahertz wave L10 emitted from the oscillating unit 61 is synchronously detected by the chopper 53, then is narrowed down by the aperture, and enters the sample supply unit 66 as a parallel wave. The scattered wave L11 generated by irradiating the sample with the terahertz wave L10 is reflected inside the integrating sphere 67 and detected by the scattered wave detection unit 68. Further, the transmitted light L12 generated by irradiating the sample with the terahertz wave L10 passes through the integrating sphere 67 and is detected by the transmitted light detection unit 69. The scattered wave detector 68 receives the scattered wave L11 and generates a voltage having a magnitude corresponding to the intensity of the scattered wave L11. In addition, the transmitted light detection unit 69 receives the transmitted light L12 and generates a voltage having a magnitude corresponding to the intensity of the transmitted light L12.

  In the detection apparatus 60 shown in FIG. 10, the oscillation unit 61 has a quantum cascade laser light source (oscillation wavelength: 4.57 μm) as a light source, and the sample supply unit 54 has a transmission cell [optical path length: 50 μm, thickness. : 80 μm], DLATAGS (deuterated L-alanine doped triglycine sulfate crystal) pyroelectric detector was used as the scattered wave detector 68 and the transmitted light detector 69.

(3) Measurement of scattered wave and transmitted light The sample obtained in Example 3 (1) was placed in the sample supply section 66 of the detection device 60 obtained in Example 3 (2). The sample in the sample supply unit 66 was irradiated with terahertz waves, and the voltage based on the scattered waves of the cells contained in the sample and the voltage based on the transmitted light were measured.

  Further, the same operation as described above was performed except that distilled water was used instead of using the sample obtained in Example 3 (1), and the voltage based on the transmitted light and the voltage based on the scattered wave were measured. Next, the scattering intensity and the transmission intensity were obtained using the voltage value based on the transmitted light and the voltage value based on the scattered wave. Thereafter, the scattering intensity / transmission intensity was determined using the scattering intensity and the transmission intensity. FIG. 11 shows the result of examining the relationship between the type of test sample and the scattering intensity / transmission intensity. In the figure, 1 indicates the scattering intensity / transmission intensity when the sample obtained in Example 3 (1) is used, and 2 indicates the scattering intensity / transmission intensity when distilled water is used.

  From the results shown in FIG. 11, the scattering intensity / transmission intensity when the sample obtained in Example 3 (1) is used is the scattering intensity / transmission intensity when distilled water is used. It can be seen that it is significantly higher than the scattering intensity / transmission intensity. From the result shown in FIG. 11 and the result obtained in Example 1, the scattered wave emitted from the detection target by irradiating the terahertz wave having a wavelength larger than the maximum diameter of the detection target such as a cell, It can be seen that the detection target contained in the sample can be detected. Therefore, by detecting the scattered wave emitted from the detection target by irradiating the dispersion containing the detection target with a terahertz wave having a wavelength larger than the maximum diameter of the detection target, the detection target included in the dispersion It is suggested that an object can be detected.

Example 4
(1) Preparation of sample Powdered milk (average particle size: 100 μm) was suspended in distilled water to obtain a dispersion as a sample.

(2) Measurement of scattered wave and transmitted light In Example 3 (3), instead of using the sample obtained in Example 3 (1), the sample obtained in Example 4 (1) was used. Except for the above, the same operation as in Example 3 (3) was performed, and the voltage based on the scattered wave of the milk powder contained in the sample and the voltage based on the transmitted light were measured. Further, in the above, except that distilled water was used instead of using the sample obtained in Example 4 (1), the same operation as described above was performed, and the voltage based on the transmitted light and the voltage based on the scattered wave Was measured. Next, the scattering intensity and the transmission intensity were obtained using the voltage value based on the transmitted light and the voltage value based on the scattered wave. Thereafter, the scattering intensity / transmission intensity was determined using the scattering intensity and the transmission intensity. FIG. 12 shows the results of examining the relationship between the type of test sample and the scattering intensity / transmission intensity. In the figure, 1 indicates the scattering intensity / transmission intensity when the sample obtained in Example 4 (1) is used, and 2 indicates the scattering intensity / transmission intensity when distilled water is used.

  From the results shown in FIG. 12, the scattering intensity / transmission intensity when the sample obtained in Example 4 (1) is used is the scattering intensity / transmission intensity when distilled water is used. It can be seen that it is significantly higher than the scattering intensity / transmission intensity.

Example 5
In the third embodiment, the same operation as in the third embodiment is performed except that the detection apparatus 1 shown in FIG. 1 having the sample supply unit 14a shown in FIG. 2 is used instead of using the detection apparatus 60 shown in FIG. To detect the detection target cell. As a result, as in Example 3, the detection target contained in the sample can be detected by the scattered wave emitted from the detection target by irradiating the terahertz wave having a wavelength larger than the maximum diameter of the detection target. I understand that I can do it. Therefore, by detecting the scattered wave emitted from the detection target by irradiating the dispersion containing the detection target with a terahertz wave having a wavelength larger than the maximum diameter of the detection target, the detection target included in the dispersion It is suggested that an object can be detected.

  As described above, according to the detection target object detection method and detection apparatus according to an embodiment of the present invention, the detection target object can be detected satisfactorily without staining the detection target object included in the dispersion. can do. For example, milk is known to increase the number of somatic cells in raw milk when the cow develops mastitis. Conventionally, bovine mastitis has been examined by staining somatic cells with a fluorescently labeled antibody or the like and detecting the fluorescence of the stained somatic cells by flow cytometry. On the other hand, according to the method of this example, terahertz waves having a wavelength larger than the maximum diameter of somatic cells are applied to milk, which is a dispersion of somatic cells and milk components such as lipids and proteins. Irradiates and measures scattered waves emitted from somatic cells, so that somatic cells contained in milk can be detected separately from the milk components without staining somatic cells with fluorescently labeled antibodies as in the past. It is suggested that you can. Therefore, it is considered that the milk used for the inspection can be used as it is for other purposes (other inspections and the like). As described above, according to the detection object detection method and the detection apparatus according to the embodiment of the present invention, it is not necessary to stain the detection object included in the sample that is the dispersion. It can be used for other purposes as it is. Therefore, the detection target object detection method and detection apparatus according to an embodiment of the present invention can be suitably used for examination of reagents, foods, etc., acquisition of information for assisting diagnosis of animal diseases, and the like. Is suggested.

DESCRIPTION OF SYMBOLS 1 Detection apparatus 2 Detection apparatus 11 Light source 12 Beam splitter 13 Oscillation part 14 Sample supply part 14a Sample supply part 14c Sample supply part 15 Detection part 15a1, 15a2 Detection part 15b1, 15b2, 15b3, 15b4 Detection part 16 Control part 17 Output part 18 Time delay unit 50 Detector 51 Oscillator 51 Transmitter 52 Lens 53 Chopper 54 Sample supply unit 55 Integrating sphere 56 Scattered wave detector 57 Transmitted light detector 60 Detector 61 Oscillator 62 Lens 63 Chopper 64 Aperture 65a, 65b Mirror 66 Sample supply unit 67 Integrating sphere 68 Scattered wave detection unit 69 Transmitted light detection unit 101 Sample supply unit main body 102 Channel 103 Incident surface 103a Lens 103 The incident surface 104 Sample introduction channel 105 Sample discharge channel 106 Output surface 111 Sample supply unit Body 11 Flow path 113 incident surface 113b thin portion 114 emitting surface 114b thin portion 115 sample introduction channel 116 sample discharge channel

Claims (2)

  A detection object detection method for detecting a detection object contained in a dispersion in which the detection object is dispersed, wherein the dispersion is irradiated with terahertz waves having a wavelength equal to or greater than a maximum particle diameter of the detection object. A method of detecting a detection target, comprising: detecting a detection target by receiving a scattered wave of a terahertz wave irradiated on the detection target. A detection object detection device used in the detection object detection method according to claim 1,
An oscillating unit for generating a terahertz wave having a wavelength equal to or greater than the maximum particle diameter of the detection target,
A sample supply unit that holds the dispersion so that the terahertz wave generated by the oscillation unit is irradiated;
A detection unit that receives the scattered wave of the terahertz wave irradiated to the detection target and detects the detection target;
The sample supply unit is made of a transmission material that transmits the terahertz wave, and has a sample supply unit main body having an incident surface having a normal line along the traveling direction of the terahertz wave.
An apparatus for detecting an object to be detected, comprising: a flow path for irradiating the dispersion with terahertz waves while moving the dispersion in the sample supply unit main body.

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