JP2009162664A - Detection method and interaction detection method using flow path, and detection apparatus - Google Patents

Abstract

A detection method capable of detecting the state of a laminar flow containing a sample simultaneously with the detection of the sample.
A method for detecting a sample flowing in a flow path, wherein a sample flow containing the sample is sandwiched between sheath flows, and stacked vertically with respect to a wall surface of the flow path facing a detection direction. A first laminar flow forming step for forming a laminar flow in the formed state, and a laminar flow for bending the laminar flow so as to be in a laminated state parallel to the wall surface after the first laminar flow forming step Provided is a detection method for performing at least a bending step and a sample detection step for detecting the sample after the laminar flow bending step.
[Selection] Figure 1

Description

  The present invention relates to a detection method using a flow path. More specifically, a detection method for detecting a sample flowing in a flow channel, an interaction detection method for detecting an interaction between substances in the flow channel, and a detection device suitable for the detection method and the interaction detection method About.

  In recent years, biological microparticles such as cells and microorganisms, microparticles such as microbeads, etc. are allowed to flow through channels and capillaries, or channels formed on a substrate such as two-dimensional or three-dimensional plastic or glass. These are detected by physical means, optical means, etc., and analyzed, separated, etc., or the interaction or reaction between substances is advanced in the flow path, and these are made physical means, optical means, etc. The technology to detect by this is advancing.

  The technology includes disease diagnosis, compound screening for drugs, forensic medicine, comprehensive analysis of genetic information, biological substance functional analysis, proteome analysis, in vivo reaction analysis, food field, agriculture field, engineering field, criminal insight field, etc. Then, it is already becoming an important core technology. A typical example of a technique for analyzing or separating fine particles using such a flow path is an analysis technique called flow cytometry.

  Flow cytometry refers to the flow of microparticles to be analyzed into a fluid, forming an array of microparticles, and irradiating the aligned microparticles with laser light or the like to emit from each microparticle. This is an analysis method in which minute particles are analyzed by detecting the detected fluorescence and scattered light, and further, the minute particles are sorted based on the analysis result.

  When detecting a sample passed through a flow path or the like as in flow cytometry, generally, a laminar flow containing the sample is formed so that the detection target is detected in the laminar flow. Detection is performed by physical means or optical means while orderly transporting the cells or the like.

  The laminar flow formation method will be described more specifically by taking flow cytometry as an example. For example, in a three-way channel, rectification of a sample such as a cell called a sheath flow is performed on both sides of the channel. A fluid medium for urging is introduced at a constant flow rate, and a sample flow containing cells or the like to be detected is injected at a constant flow rate into the central channel of the three-way channel. At this time, since the Reynolds number in the flow path is relatively small, the flows are not mixed with each other due to the principle of laminar flow (laminar flow) in the flow path, and a stratified flow (laminar flow) is formed. The

  As described above, in order to accurately detect and sort the sample passed through the flow path or the like, it is important how the laminar flow can be controlled. For example, in Patent Document 1, as a microchip capable of controlling the movement direction of fine particles in order to easily and accurately separate fine particles such as cells, a fine particle-containing solution introduction channel and at least one of the channels are provided on a substrate. A sheath flow forming channel disposed on one side, a fine particle measurement site for measuring the introduced fine particles, and two or more fine particles for separately collecting fine particles placed downstream of the fine particle measurement site There is disclosed a microparticle sorting microchip having a sorting channel and two or more electrodes for controlling the moving direction of the microparticles installed in the vicinity of the channel opening from the microparticle measurement site to the microparticle sorting channel.

  In Patent Document 2, as a method for efficiently controlling or focusing the flow of a microfluidic fluid, a sample fluid is flowed in a central channel (flow path), a sheath flow is flowed in a focusing channel (flow path), and A method of controlling or focusing the flow of sample fluid by controlling the flow rate of the sheath fluid is disclosed.

Furthermore, in Patent Document 3, as a technique for stabilizing an unstable flow (laminar flow), a technique using a sheath flow to which a viscosity increasing agent is added in order to give the sheath flow a viscosity higher than that of water. Has proposed.
JP 2003-107099 A JP 2004-93553 A JP-T-2004-500562

  In order to accurately detect and sort a sample flowing through a flow path etc., it is important whether or not the laminar flow can be controlled. As mentioned above, the conventional technology for controlling the laminar flow And techniques for stabilizing laminar flow have been developed.

  However, in the conventional method, before passing the target sample through the flow path, the reference spectrum beads or reference diameter beads corresponding to the target sample are passed through the flow path in advance to obtain the optimum laminar flow. The position, the amount of laminar flow, the light irradiation position, the light receiving position, the light source power, the applied voltage and the like need to be determined, which is complicated.

  Further, in the flow channel, the width and position of the laminar flow may change during detection due to the influence of the flow resistance on the flow channel wall side, the surface tension of the flow channel wall, and the like. In addition, samples with different sizes and properties are often present in the sample liquid, so the optimum laminar flow position, laminar flow amount, light irradiation position, light receiving position, and light source are suitable for the target sample during detection. There are cases where it is desired to change the power, applied voltage, and the like.

  In this case, since the conventional method cannot detect the state of the laminar flow containing the sample during detection, the position of the laminar flow, the amount of laminar flow, the light irradiation position, the light receiving position, and the light source are detected during detection. It was difficult to control power, applied voltage, and the like.

  Further, conventionally, it has been difficult to control the laminar flow shape of the fluid flowing in the flow path in a direction parallel or perpendicular to the detection direction, and it has been difficult to perform highly accurate detection.

  Therefore, the main object of the present invention is to provide a detection method capable of detecting information from a laminar flow containing a sample simultaneously with detection of the target sample.

In the present invention, first, a method for detecting a sample flowing in a flow path,
A first laminar flow forming step of forming a laminar flow in a state of being stacked perpendicular to the wall surface of the flow channel facing the detection direction by sandwiching a sample flow containing the sample with a sheath flow;
After passing through the first laminar flow forming step, the laminar flow bending step of bending the laminar flow so as to be in a laminated state parallel to the wall surface;
A sample detection step of detecting the sample after the laminar flow bending step;
A detection method for performing at least the above is provided.
In the detection method according to the present invention, a laminar flow information detecting step of detecting a laminar flow width and / or position of the laminar flow formed in the first laminar flow forming step after the first laminar flow forming step is performed. It is also possible to do this.
In the sample detection step, it is more preferable to detect the sample based on the laminar flow information detected in the laminar flow information detection step.
In the laminar flow bending step, the method is not particularly limited as long as the laminar flow is bent so as to be in a laminated state parallel to the wall surface of the flow channel facing the detection direction. ) When the laminar flow direction is the X direction, a first bending step of bending the laminar flow approximately 90 degrees in the Y-axis direction; and (2) a first bending step of bending the laminar flow approximately 90 degrees in the Z-axis direction. If at least the two-folding step is performed, the laminar flow can be reliably bent so as to be in a laminated state parallel to the wall surface.
In the detection method according to the present invention, it is possible to perform a laminar flow control step of controlling the laminar flow width and / or position of the laminar flow before performing the sample detection step.
In the detection method according to the present invention, a detection condition control step for controlling detection conditions in the sample detection step can be performed before the sample detection step.
In the detection method according to the present invention, a laminar flow in a laminated state is formed in the first laminar flow forming step, and after the laminar flow bending step, the laminar flow is sheathed from both sides of the laminated state. It is preferable to perform the second laminar flow forming step of further sandwiching with the flow.
It is more preferable to perform a third laminar flow forming step of forming a plurality of laminar flows by combining two or more laminar flows that have undergone the second laminar flow forming step.
The specific detection method in the laminar flow information detection step and the sample detection step is not particularly limited as long as the laminar flow information and the sample information can be detected. However, for example, the detection can be performed using an optical method. is there.

In the present invention, next, a method for detecting an interaction between the substance A and the substance B flowing through the flow path,
A sample flow containing the substance A is sandwiched by a sheath flow containing at least a part of the substance B, thereby forming a laminar flow in a state of being stacked perpendicular to the wall surface of the flow channel facing the detection direction. A first laminar flow forming step;
After passing through the first laminar flow forming step, the laminar flow bending step of bending the laminar flow so as to be in a laminated state parallel to the wall surface;
An interaction detecting step for detecting an interaction between the substance A and the substance B after the laminar flow bending process;
An interaction detection method for performing at least the above is provided.
In the interaction detection method according to the present invention, laminar flow information detection for detecting the laminar flow width and / or position of the laminar flow formed in the first laminar flow forming step after the first laminar flow forming step. It is also possible to carry out further steps.
In the interaction detection step, it is more preferable to detect the interaction between the substance A and the substance B based on the laminar flow information detected in the laminar flow information detection step.
In the laminar flow bending step, the method is not particularly limited as long as the laminar flow is bent so as to be in a laminated state parallel to the wall surface of the flow channel facing the detection direction. ) When the laminar flow direction is the X direction, a first bending step of bending the laminar flow approximately 90 degrees in the Y-axis direction; and (2) a first bending step of bending the laminar flow approximately 90 degrees in the Z-axis direction. If at least the two-folding step is performed, the laminar flow can be reliably bent so as to be in a laminated state parallel to the wall surface.
In the interaction detection method according to the present invention, it is possible to perform a laminar flow control step of controlling the laminar flow width and / or position of the laminar flow before performing the interaction detection step.
In the interaction detection method according to the present invention, a detection condition control step for controlling detection conditions in the interaction detection step can be performed before the interaction detection step.
In the interaction detection method according to the present invention, a laminar flow in a laminated state is formed in the first laminar flow forming step, and further, after passing through the laminar flow bending step, the laminar flow is converted into both sides of the laminated state. It is preferable to perform the second laminar flow forming step of further sandwiching with the sheath flow.
It is more preferable to perform a third laminar flow forming step of forming a plurality of laminar flows by combining two or more laminar flows that have undergone the second laminar flow forming step.
The specific detection method in the laminar flow information detection step and the interaction detection step is not particularly limited as long as laminar flow information and the interaction between the substance A and the substance B can be detected. For example, an optical method is used. Detection can be performed.

In the present invention, further, a detection device for a sample flowing in the flow path,
When the flow direction is the X direction, a first bent portion where the flow path bends approximately 90 degrees in the Y-axis direction, a second bent portion where the flow path bends approximately 90 degrees in the Z-axis direction, A flow path comprising at least
A first detector for detecting a laminar flow width and / or position of a laminar flow containing the sample in at least an upstream flow path of the first bent portion and the second bent portion;
A second detector for detecting the sample in a flow path at least downstream of the first bent portion and the second bent portion;
Is provided.

  Here, technical terms used in the present invention are defined. The “sample” in the present invention refers to biologically related microparticles such as cells, microorganisms, liposomes, DNA, and proteins, synthetic particles such as latex particles, gel particles, and industrial particles, or liquid substances that are not limited to solids. Any substance that can flow through the flow path 1 is included.

  The “sheath flow” in the present invention refers to a fluid medium for promoting the rectification of the sample flow containing the sample, and is a concept including a substance containing some substance as long as the rectification of the sample flow is not impaired.

  In the detection method according to the present invention, it is possible to detect information of a target sample and simultaneously detect information from a laminar flow containing the sample. For this reason, it is possible to omit a preparatory step before the detection of the target sample, and it is possible to control individual detection conditions during the detection.

  DESCRIPTION OF EXEMPLARY EMBODIMENTS Hereinafter, preferred embodiments for carrying out the invention will be described with reference to the drawings. In addition, embodiment described below shows an example of typical embodiment of this invention, and, thereby, the range of this invention is not interpreted narrowly.

<Detection method>
FIG. 1 is a flowchart of a detection method according to the present invention.

  The detection method according to the present invention is a method for detecting a sample flowing in a flow path, and includes a first laminar flow forming step (I), a laminar flow bending step (II), a sample detecting step ( III) and at least. Moreover, it is possible to perform a laminar flow information detection process (IV) as needed. Hereinafter, each step will be described in detail.

(I) 1st laminar flow formation process 1st laminar flow formation process (I) is 1d of wall surfaces 1d of the flow path 1 which faces the detection direction D1 by pinching the sample flow F1 containing a sample with sheath flow F21, F22. ₁ is a step of forming a laminar flow (sample flow F1, sheath flow F21, F22) in a state of being laminated perpendicularly to 1 (see the flow diagram in FIG. 1). This will be described in detail with reference to FIG.

  FIG. 2 is a schematic perspective view of a flow channel 1 that can be used in the detection method according to the present invention.

  The form of the flow channel 1 capable of performing the detection method according to the present invention is not particularly limited as long as it can form a laminar flow, and can be freely designed. For example, the detection method according to the present invention can be performed not only in the form shown in FIG. 2 but also in the flow path 1 formed on a two-dimensional or three-dimensional plastic or glass substrate.

  Further, the channel width, the channel depth, and the channel cross-sectional shape of the channel 1 are not particularly limited as long as they can form a laminar flow, and can be freely designed. For example, the detection method according to the present invention can be performed even in a micro flow channel having a flow channel width of 1 mm or less. In particular, the detection method according to the present invention can be more suitably performed by using a microchannel having a channel width of about 10 μm to 1 mm.

The flow channel 1, to the wall surface 1d ₁ of the channel 1 facing the detecting direction D1 shown in FIG. 2 reference numerals D1, laminar flow in a state of being vertically stacked (sample flow F1, sheath flows F21, F22) Form. The laminar flow (sample flow F1, sheath flow F21, F22) is formed by sandwiching the sample flow F1 containing the sample between the sheath flows F21, F22 as shown in FIG.

Specific methods sandwiching the sample flow F1 in the sheath flow F21, F22, especially if formed laminar flow in a state of being vertically stacked with respect to the wall surface 1d ₁ of the channel 1 (sample flow F1, sheath flows F21, F22) Without limitation, any known method can be adopted. As a preferred example, as shown in FIG. 3, a sample flow channel 11 and a sheath flow channel 12 are provided in the flow channel 1, and the sheath flow channel 12 is joined from both sides of the sample flow channel 11. Thus, the sample flow F1 introduced from the sample flow passage 11 can be sandwiched from both sides by the sheath flows F21 and F22 introduced from the sheath flow passage 12.

(II) Laminar flow folding process The laminar flow folding process (II) faces the laminar flow (sample flow F1, sheath flow F21, F22) in the detection direction D2 after passing through the first laminar flow formation step (I). This is a step of bending so as to be in a laminated state parallel to the wall surface 1d _{2 of the} flow path 1 (see the flowchart in FIG. 1).

Laminar flow (sample flow F1, sheath flows F21, F22), by a parallel stacked against the wall surface 1d ₂ of the channel 1 facing the detection direction D2, preferably a sample detection step (III) described below Can be done. This will be described in detail with reference to FIG.

  FIG. 4 is a diagram for comparing the inside of the flow path 1 before and after the laminar flow bending step (II), and the diagram indicated by reference numeral (i) in FIG. 4 is a flow path viewed from the detection directions D1 and D2 in FIG. 1 is a schematic cross-sectional view showing the flow path 1 viewed from the side S1 and S2 with respect to the detection directions D1 and D2, and is indicated by the reference sign (i-1). ) And (ii-1) are schematic sectional views showing the flow path 1 before the laminar flow bending step (II), respectively, and are shown by reference numerals (i-2) and (ii-2). These are the cross-sectional schematic diagrams which show the flow path 1 after performing each laminar flow bending process (II).

As shown in FIG. 4 (i-1), before the laminar flow bending step (II), the laminar flow (sample flow F1, sheath flow F21, F22) is the wall surface 1d of the flow channel 1 facing the detection direction D1. ₁ , the laminar flow width (sample flow width f1, sheath flow width) of each laminar flow (sample flow F1, sheath flow F21, F22) when viewed from the detection direction D1. f21, f22) and position can be detected accurately.

  However, as shown in the diagram viewed from the side S1 in FIG. 4 (ii-1), even if it is attempted to detect the information of the sample C in the sample flow F1 from the detection direction D1, a plurality of samples C overlap on the detection axis D11. Therefore, it is difficult to separately detect information from the sample C with high accuracy.

Therefore, by performing the laminar flow bending step (II), as shown in FIG. 4 (ii-2), the laminar flow (sample flow F1, sheath flow F21, F22) is a flow path 1 facing the detection direction D2. with parallel stacked against the wall surface 1d ₂ of, for prevented that the sample C overlap more on detection axes on D21, the sample detection step described later (III), exactly the information from the sample C It becomes possible to detect.

  Further, the laminar flow width (sample flow width f1, sheath flow widths f21, f22) of the laminar flow (sample flow F1, sheath flow F21, F22) obtained in the laminar flow information detection step (IV) described later, and the flow path 1 After the laminar flow bending process (II), the position in FIG. 4 becomes position information in the depth direction in the flow path 1 viewed from the detection direction D2, and therefore information from the sample C in the sample detection process (III) described later. Can be detected with high accuracy.

  In addition, after performing the laminar flow bending process (II), when viewed from the detection direction D2, as shown in FIG. It is difficult to detect the laminar flow width (sample flow width f1, sheath flow width f21, f22) and position of each laminar flow (sample flow F1, sheath flow F21, F22).

  In this way, the laminar flow folding step (II) is provided, and each layer is detected before and after the laminar flow folding step (II). In the flow information detection step (IV), the laminar flow width (sample flow width f1, sheath flow width f21, f22) and position of each laminar flow (sample flow F1, sheath flow F21, F22) can be detected. After the bending step (II), information from the sample C can be accurately detected in the sample detection step (III) described later.

In laminar flow bending step (II), flow layers (sample flow F1, sheath flows F21, F22), is bent so as to be parallel stacked against the wall surface 1d ₂ of the channel 1 facing the detection direction D2 If it is, the method is not specifically limited, It can be bent freely using arbitrary methods.

For example, as shown in FIG. 2 and FIG. 3, (1) when the flow direction of the laminar flow (sample flow F1, sheath flow F21, F22) is the X direction, the laminar flow (sample flow F1, sheath flow F21, F22). ) Is bent approximately 90 degrees in the Y-axis direction, and (2) the second bending process is bent approximately 90 degrees in the Z-axis direction of the laminar flow (sample flow F1, sheath flow F21, F22). if at least performed, flow layers (sample flow F1, sheath flows F21, F22) and, certainly, can be parallel stacked against the wall 1d _2.

  The order in which the first bending step (1) and the second bending step (2) are performed is not particularly limited, and the second bending step (2) may be performed after the first bending step (1). You may perform a 1st bending process (1) after performing a bending process (2).

  Further, in the first bending step (1), if the laminar flow (sample flow F1, sheath flow F21, F22) can be bent in the Y-axis direction, as shown in FIG. 5 and FIG. Even if it is this bending, it can be set as a lamination | stacking state parallel to wall surface 1d2 by performing a 2nd bending process (2) after that.

As shown in FIG. 7, (1) a first bending step of bending a laminar flow (sample flow F1, sheath flow F21, F22) approximately 90 degrees in the Y-axis direction, and (2) laminar flow (sample flow F1). , The second bending step of bending the sheath flows F21, F22) in the Z-axis direction by approximately 90 degrees, and (3) the second bending process of bending the laminar flows (sample flow F1, sheath flows F21, F22) by approximately 90 degrees in the X-axis direction. by performing a two bending steps, at least, a flow layer (sample flow F1, sheath flows F21, F22), it may be a parallel stacked against the wall surface 1d _2. In this case, since the entire flow channel 1 is formed on substantially the same axis, especially when the flow channel 1 is formed on a substrate such as a two-dimensional or three-dimensional plastic or glass, the area is limited. This is effective.

  In the illustration of the laminar flow bending method, all of the first bending step (1), the second bending step (2), and the third bending step (3) are laminar flows (sample flow F1, sheath flow F21, F22) is bent at a substantially right angle, but about 90 ° ± 20 ° is acceptable. Further, the present invention is not limited to this, and for example, as shown in FIG. 8, it is possible to bend the laminar flow (sample flow F1, sheath flow F21, F22) by making the flow path 1 substantially spiral. .

  Although not illustrated, the laminar flow bending step (II) described above can also be performed in the flow path 1 formed on a two-dimensional or three-dimensional plastic or glass substrate. In this case, in the laminar flow bending step (II), the laminar flow (sample flow F1, sheath flow F21, F22) in a state of being laminated perpendicularly to the substrate is brought into a lamination state parallel to the substrate. It will be bent.

(III) Sample Detection Step The sample detection step (III) is a step of detecting information from the sample C after passing through the laminar flow bending step (II) (see the flowchart in FIG. 1).

  In the sample detection step (III), the detection method is not particularly limited as long as the sample C contained in the sample flow F1 can be detected, and a known method can be freely adopted. For example, an optical method, an electrical method, a magnetic method, etc. can be mentioned. In particular, in the present invention, an optical method is suitable.

  Examples of optical methods include fluorescence measurement, scattered light measurement, transmitted light measurement, reflected light measurement, diffracted light measurement, ultraviolet spectroscopic measurement, infrared spectroscopic measurement, Raman spectroscopic measurement, FRET measurement, FISH measurement, and various other spectrum measurements. The method is mentioned. Further, if detection is performed using an area imaging device such as a charge coupled device (CCD) or a complementary metal oxide semiconductor (CMOS), an image emitted from the sample C on the entire screen can be photoelectrically converted.

  As a specific example, light irradiation is performed from the direction of arrow D2 in FIG. 2, and fluorescence or scattered light generated from the sample C in the sample flow F1 by the light irradiation is detected by a photodetector. Sample C can be detected.

  The type of light irradiation when the optical method is employed is not particularly limited, but light having a constant light direction, wavelength, and light intensity is desirable in order to reliably generate fluorescence and scattered light from the sample. As an example, a laser or LED (Light Emission Diode) can be mentioned.

  In addition, the light irradiation method in the case of adopting the optical method is not particularly limited. For example, as shown by the symbol W2 in FIG. 2, optical information is detected while scanning the irradiation spot in the width direction of the flow path 1. Alternatively, as indicated by reference numeral L2 in FIG. 2, the optical information may be detected while the irradiation spot is scanned in the flow path 1 flow direction.

  Examples of the electrical method include a method of measuring a resistance value, a capacitance value (capacitance value), an inductance value, an impedance, a change value of an electric field between electrodes, and the like regarding the sample C contained in the sample flow F1.

  Examples of the magnetic method include a method in which a magnetic material is modified on the surface of the sample C contained in the sample flow F1, and measurement of magnetization, magnetic field change, magnetic field change, and the like is performed.

  In addition, when performing the laminar flow information detection process (IV) mentioned later, the laminar flow width (sample flow F1) of the laminar flow (sample flow F1, sheath flow F21, F22) detected in the laminar flow information detection step (IV) It is desirable to detect the sample C based on the width f1, the sheath flow width f21, f22) and / or the position information. In the laminar flow information detection step (IV), the laminar flow width (sample flow width f1, sheath flow width f21, f22) of the laminar flow (sample flow F1, sheath flow F21, F22) and position information in the flow path 1 are detected. This is because the sample C contained in the sample flow F1 can be reliably detected with high accuracy.

(IV) Laminar Flow Information Detection Step The laminar flow information detection step (IV) is the laminar flow width and flow of the laminar flow (sample flow F1, sheath flow F21, F22) formed in the first laminar flow formation step (I). This is a step of detecting the position on the road 1 (see the flowchart in FIG. 1).

  Specifically, in the first laminar flow forming step (I), the laminar flow width of the laminar flow (sample flow F1, sheath flow F21, F22) formed in the laminated state and the position in the flow path 1 are set in the laminated state. It is detected from the cross-section side (see symbol D1 in FIG. 2).

  In the laminar flow information detection step (IV), the detection method is not particularly limited as long as the laminar flow width of the laminar flow (sample flow F1, sheath flow F21, F22) and the position in the flow path 1 can be detected. Can be freely adopted. For example, an optical method, an electrical method, a magnetic method, etc. can be mentioned. In particular, in the present invention, an optical method is suitable.

  Examples of optical methods include fluorescence measurement, scattered light measurement, transmitted light measurement, reflected light measurement, diffracted light measurement, ultraviolet spectroscopic measurement, infrared spectroscopic measurement, Raman spectroscopic measurement, FRET measurement, FISH measurement, and various other spectrum measurements. The method is mentioned. Further, if detection is performed using an area imaging device such as a charge coupled device (CCD) or a complementary metal oxide semiconductor (CMOS), an image emitted from the channel 1 of the entire screen can be photoelectrically converted.

  As a specific example, light or light is emitted from the direction of the arrow D1 in FIG. 2, and fluorescence or scattered light generated from a material such as the sample C contained in the sample flow F1 or the sheath flows F21 and F22 by the light irradiation. Can be detected by the photodetector to detect the laminar flow width of the sample flow F1 and the sheath flows F21 and F22 and the position in the flow path 1.

  The type of light irradiation when the optical method is employed is not particularly limited, but light having a constant light direction, wavelength, and light intensity is desirable in order to reliably generate fluorescence and scattered light from the sample. As an example, a laser or LED (Light Emission Diode) can be mentioned.

  In addition, the light irradiation method in the case of adopting the optical method is not particularly limited. For example, as shown by the symbol W1 in FIG. 2, even if the optical information is detected while scanning the irradiation spot in the flow path width direction. Good.

  As an electrical method, for example, a resistance value, a capacitance value (capacitance value), an inductance value, an impedance, a change value of an electric field between electrodes, and the like related to a substance such as the sample C contained in the sample flow F1 and the sheath flows F21 and F22. The method of measuring is mentioned.

  Examples of the magnetic method include a method in which a magnetic material is modified on the surface of a substance such as the sample C contained in the sample flow F1 or the sheath flows F21 and F22 to measure magnetization, magnetic field change, magnetic field change, and the like. It is done.

Thus, in the detection method according to the present invention, by performing the first layer flow forming step (I), in a state of being vertically stacked with respect to the wall surface 1d ₁ of the channel 1 facing the detection direction D1 In order to form a laminar flow (sample flow F1, sheath flow F21, F22), laminar flow width and flow path of laminar flow sample flow F1, sheath flow F21, F22) in the subsequent laminar flow information detection step (IV) The position in 1 can be accurately detected.

  The detection method according to the present invention performs at least the first laminar flow forming step (I), the laminar flow bending step (II), and the sample detection step (III) described above, and if necessary, a laminar flow information detection step (IV) is a method for performing the following laminar flow control step (V), detection condition control step (VI), second laminar flow formation step (VII), and third laminar flow formation step (VIII). It can be performed as needed. Hereinafter, each step will be described in detail.

(V) Laminar Flow Control Step The laminar flow control step (V) is performed before the sample detection step (III) is performed. The laminar flow width (sample flow width f1, This is a step of controlling the sheath flow width f21, f22) and / or the position in the flow path 1 (see the flow chart in FIG. 1).

  The laminar flow control step (V) may be performed before the sample detection step (III). For example, as shown in the flowchart of FIG. 1, the laminar flow control step (V) may be performed before the laminar flow bending step (II). It may be performed after the flow folding process (II). However, when the laminar flow information detection step (IV) is performed, it is preferable that the laminar flow information detection step (IV) is performed. This is because laminar flow control can be performed based on the laminar flow information detected in the laminar flow information detection step (IV). Hereinafter, a specific example of the laminar flow control method will be described with reference to FIGS.

  FIG. 9 is a schematic cross-sectional view of the laminar flow (sample flow F1, sheath flow F21, F22) after undergoing the laminar flow information detection step (IV) as viewed from the side of the laminated state. The figure shown by i) shows the state before the laminar flow control step (V), and the figure shown by the symbol (ii) shows the state after the laminar flow control step (V).

  In the laminar flow information detection step (IV), as shown in FIG. 9 (i), the sheath flow width f 21 is narrower than the sheath flow width f 22, and the position of the sample flow F 1 is shifted from the center of the flow path 1. Is detected, for example, by increasing the feeding amount of the sheath flow F21, as shown in FIG. 9 (ii), the sheath flow width f21 and the sheath flow width f22 are made substantially the same, and the sample flow F1 Can be controlled toward the center of the flow path 1.

  In the present embodiment, the position of the sample flow F1 is controlled to the central portion of the flow path 1. However, the present invention is not limited to this, and the liquid feed amounts of the sample flow F1, sheath flow F21, and F22 are adjusted as necessary. The position of the sample flow F1 can be freely controlled to an arbitrary position in the flow path 1 by increasing or decreasing.

  FIG. 10 is a schematic cross-sectional view of a laminar flow (sample flow F1, sheath flow F21, F22) different from FIG. 9 after undergoing the laminar flow information detection step (IV), as viewed from the side of the laminated state. In FIG. 10, the diagram indicated by reference numeral (i) shows the state before the laminar flow control step (V) is performed, and the diagram indicated by reference numeral (ii) shows the state after the laminar flow control step (V).

  In the laminar flow information detection step (IV), when it is detected that the flow path width f1 of the sample flow F1 is larger than the particle size of the sample C as shown in FIG. The detection accuracy in the sample detection step (III) may be reduced because two or more samples C may flow in the direction in which the sample C is overlapped, or the order of the samples C may be changed. There is.

  Therefore, by increasing the amount of the sheath flows F21 and F22 fed, the laminar flow (sample is set so that the flow path width f1 of the sample flow F1 matches the particle size of the sample C as shown in FIG. 10 (ii). By controlling the flow F1 and the sheath flows F21 and F22), for example, when the sample C is optically detected, the samples C such as cells and fine particles may be arranged one by one in the detection spot. In addition, the detection accuracy in the sample detection step (III) can be improved such that detection can be performed separately.

  FIG. 11 is a schematic cross-sectional view of a laminar flow (sample flow F1, sheath flow F21, F22) different from that of FIGS. 9 and 10 after the laminar flow information detection step (IV), as viewed from the side of the laminated state. In FIG. 11, the diagram indicated by reference sign (i) shows the state before the laminar flow control step (V) is performed, and the diagram indicated by reference sign (ii) shows the state after the laminar flow control step (V) is performed. Yes. A broken line curve indicated by reference sign V in FIG. 11 is a Hagen-Poiseuille velocity curve indicating a fluid velocity distribution in the flow path 1.

  In the flow channel 1, the flow velocity at the center of the flow channel 1 is the fastest as shown by the fluid velocity distribution curve V due to the influence of flow resistance on the flow channel wall side, surface tension of the flow channel wall, and the like. As you get closer to the wall, the flow velocity decreases.

  Therefore, as shown in FIG. 11 (i), when the sample flow F1 flows through the central portion of the flow path 1, the flow rate of the sample C in the sample flow F1 also increases, and the detection sensitivity in the sample detection step (III) is increased. There is a case where it falls.

  Therefore, for example, as shown in FIG. 11 (ii), the sample flow F1 is biased toward the sheath flow F21 by decreasing the amount of the sheath flow F21 and increasing the amount of the sheath flow F22. By controlling the flow rate of the sample flow F1, the detection sensitivity in the sample detection step (III) can be improved.

  In the present embodiment, control is performed to reduce the flow rate of the sample flow F1, but the present invention is not particularly limited to this. For example, in order to increase the detection speed in the sample detection step (III), the sample flow F1 is flowed. It is also possible to increase the flow rate of the sample flow F1 by controlling the flow through the center of the path 1.

  Such fluid velocity control can also be realized by the method shown in FIG. FIG. 12 is a schematic cross-sectional view of a laminar flow (sample flow F1, sheath flow F21, F22) different from FIGS. 9 to 11 after the laminar flow information detection step (IV) is viewed from the side of the laminated state. In FIG. 12, the diagram indicated by reference numeral (i) shows the state before the laminar flow control step (V) is performed, and the diagram indicated by reference numeral (ii) shows the state after the laminar flow control step (V) has been performed. Yes.

  In general, when a fluid is introduced into the flow path 1 at the same pressure, the flow velocity is reduced to 1/4 times when the flow path width H is doubled. Therefore, as shown in FIG. 12 (ii), the flow velocity V of the sample flow F1 can be controlled to be reduced to V / 4 by doubling the flow path width H, and the sample detection step (III ) Can be improved.

  In the present embodiment, control is performed to reduce the flow rate of the sample flow F1, but the present invention is not limited to this. For example, in order to increase the detection speed in the sample detection step (III), the flow path width H is set to It is also possible to control such that the flow velocity V of the sample flow F1 is increased by narrowing. Further, not only the flow path width H but also the flow velocity V of the sample flow F1 can be changed by changing the depth and cross-sectional shape of the flow path.

(VI) Detection Condition Control Step The detection condition control step (VI) is a step of controlling the detection conditions in the sample detection step (III) before performing the sample detection step (III) (see the flowchart in FIG. 1).

  The detection condition control step (VI) may be performed at least before performing the sample detection step (III). For example, as shown in the flow chart of FIG. 1, the detection condition control step (VI) may be performed before performing the laminar flow bending step (II). It may be performed after the laminar flow folding step (II). Although not shown, the second laminar flow forming step (VII) and the third laminar flow forming may be performed before the second laminar flow forming step (VII) and the third laminar flow forming step (VIII) described later. You may perform after performing process (VIII). However, when the laminar flow information detection step (IV) is performed, it is preferable that the laminar flow information detection step (IV) is performed. This is because detection condition control can be performed based on the laminar flow information detected in the laminar flow information detection step (IV).

  In the detection condition control step (VI), the detection conditions in the sample detection step (III) are controlled, but the detection conditions for performing the control are not particularly limited. For example, the position of the light irradiation spot, the shape of the irradiation spot, the light irradiation intensity, the pulse width, the pulse ratio, the focal position, the amplification factor of the photodetector, the amplification factor of the light irradiation device, and the like can be controlled.

  An example of detection condition control will be described with reference to FIG. 13 is a schematic cross-sectional view of the laminar flow (sample flow F1, sheath flow F21, F22) after undergoing the laminar flow information detection step (IV) as viewed from the side of the laminated state. The figure shown by i) shows the state before performing the detection condition control step (VI), and the figure shown by the symbol (ii) shows the state after performing the detection condition control step (VI).

  For example, in the laminar flow information detection step (IV), as shown in FIG. 13 (i), the sheath flow width f21 is narrower than the sheath flow width f22, and the position of the sample flow F1 is biased toward the sheath flow F21. 13 (ii), the detection accuracy in the sample detection step (III) can be improved by controlling the position of the light irradiation spot to the flow position of the sample flow F1. it can.

  Conventionally, when the sample C flowing through the flow channel 1 is detected, a laminar flow (sample flow F1) including the target sample C is formed at the center of the flow channel 1, and light is directed toward the sample C. Irradiation and the like were performed, and sample C was detected. However, in this method, only information from the sample C can be obtained during the detection, and therefore it is difficult to control the detection conditions optimal for the detection during the detection. For this reason, a preparatory step in which a reference spectrum bead or a reference diameter bead corresponding to the sample C is preliminarily distributed and the optimum detection condition for the detection is specified, and then the sample C is actually passed to perform the detection. It was necessary.

  However, in the detection method according to the present invention, the laminar flow information detection step (IV) is performed in a state where the sample C is actually passed through the flow path 1, and this information can be used for detection condition control. It is not necessary to perform a preparatory step for distributing reference spectrum beads, reference diameter beads and the like corresponding to the sample C in advance. Therefore, the detection time can be shortened and the detection accuracy can be improved.

(VII) Second laminar flow forming step In the second laminar flow forming step (VII), the laminar flow (sample flow F1, sheath flow F21, F22) is laminated after the laminar flow bending step (II). This is a step of further sandwiching the sheath flows F23 and F24 from both sides of the state (see the flowchart in FIG. 1). This will be described in detail with reference to FIGS.

  14 and 15 are schematic perspective views of the flow channel 1 different from those shown in FIGS. 2, 3, and 5 to 8 that can be used in the detection method according to the present invention.

After passing through the laminar flow bending step (II), the flow channel 1 is laminated in parallel with the wall surface 1d ₂ of the flow channel 1 facing the detection direction D2 indicated by reference numeral D2 in FIGS. Laminar flow (sample flow F1, sheath flow F21, F22) is formed. When further sandwiched by the sheath flows F23 and F24 from both sides of the laminated state, the sample flow F1 forms a laminar flow in a state where the sample flow F1 is converged at the central portion of the flow path 1, as shown in FIGS. be able to.

  A specific method of sandwiching the laminar flow (sample flow F1, sheath flow F21, F22) between the sheath flows F23, F24 is as long as the sample flow F1 can form a laminar flow in a state where the sample flow F1 is converged at the center of the flow path 1. It does not specifically limit and all well-known methods are employable. As a preferred example, as shown in FIGS. 14 and 15, the sample flow F <b> 1 is made to flow by joining the sheath flow passage 13 from both sides of the flow passage 1 after the laminar flow bending step (II). It is possible to form a laminar flow that is converged at the center of one.

  In this way, by performing the second laminar flow forming step (VII), if the sample flow F1 is formed in a converged state at the center of the flow path 1, it is viewed from either the detection direction D2 or the side S2. Even in this case, the sample C can be detected in an orderly manner.

(VIII) Third Laminar Flow Formation Step In the third laminar flow formation step (VIII), two or more laminar flows (sample flow F1, sheath flow F21, F22) that have undergone the second laminar flow formation step (VII) are merged. This is a step of forming a plurality of laminar flows (sample flow F1, sheath flows F21, F22) (see the flow chart in FIG. 1). This will be described in detail with reference to FIG.

  16 is a top schematic plan view of the flow channel 1 formed on the substrate T that can be used in the detection method according to the present invention, and the diagram indicated by reference numeral (ii) It is a cross-sectional schematic diagram of the flow path 1 of the broken-line circle | round | yen part shown by code | symbol R1 in FIG.16 (i). Reference numeral 111 denotes a sample flow inlet, and reference numerals 121 and 131 denote sheath flow inlets, respectively. In the present embodiment, the flow path 1 is provided on the substrate T. However, the present invention is not particularly limited to this configuration. For example, a flow path in which a plurality of flow paths 1 shown in FIG. 14 or FIG. Is also possible.

  After passing through the second laminar flow formation step (VII), the flow path 1 has a laminar flow (sample flow F1, sheath flows F21, F22, F23, F24) is formed (see FIG. 16 (ii)). When two or more laminar flows (sample flow F1, sheath flows F21, F22, F23, F24) are merged as shown in FIG. 16 (i), a plurality of laminar flows (sample flow F1, sheaths as shown in FIG. 17) are obtained. Streams F21, F22, F23, F24) are formed.

  The diagram indicated by reference numeral (i) in FIG. 17 is a schematic cross-sectional view of the flow path 1 of the broken-line circle portion indicated by reference numeral R2 in FIG. 16I viewed from the detection direction D2, and is indicated by reference numeral (ii). These are the cross-sectional schematic diagrams which looked at the flow path 1 of the broken-line circle | round | yen part shown by the code | symbol R2 in FIG.16 (i) from the flow direction.

  When the third laminar flow forming step (VIII) is performed, a plurality of sample flows F1 can be formed as viewed from the detection direction D2, as shown in FIG. Therefore, for example, by changing the type of the sample C contained in each sample flow F1, many types of the sample C can be detected at a time, and the detection time can be greatly shortened.

  Sample C that can be detected by the detection method described above is a cell-related microparticle such as a microorganism, liposome, DNA, protein, or synthetic particle such as latex particle, gel particle, industrial particle, or solid. Any material that can flow through the flow path 1 such as a liquid material is included.

<Interaction detection method>
An interaction detection method according to the present invention is a method for detecting an interaction between a substance A and a substance B flowing through a flow path, and includes a first laminar flow forming step (I), a laminar flow bend This is a method of performing at least step (II) and interaction detection step (III). Further, a laminar flow information detection step (IV) can be performed as necessary. Furthermore, if necessary, a laminar flow control step (V), a detection condition control step (VI), a second laminar flow forming step (VII), and a third laminar flow forming step (VIII) can be performed. Since each process is the same as each process of the detection process mentioned above, the item peculiar to an interaction detection method is demonstrated using FIG.18 and FIG.19 below.

  FIG. 18 is a view for explaining a state in the flow channel 1 when performing the interaction detection method according to the present invention, and laminar flow (sample flow F1, sheath flow F21, F22) is placed on the side in the stacked state. It is the cross-sectional schematic diagram seen from the direction.

  In the interaction detection method according to the present invention, the substance A is contained in the sample flow F1, the substance B is contained in at least one of the sheath flows F21, F22, and the interaction that progresses between these substances A and B is detected. . In the present embodiment, the substance B is contained in the sheath flow F21. However, the present invention is not limited to this. Even if the substance B is contained in the sheath flow F22, the substance B is contained in both the sheath flow F21 and the sheath flow F22. May be included.

  It is also possible to detect different interactions simultaneously by containing different types of substances B in the sheath flow F21 and the sheath flow F22.

  The substance A is not particularly limited as long as it is a substance that can flow through the flow path 1. Examples thereof include living body-related microparticles such as cells, microorganisms, liposomes, DNA, and proteins, or synthetic particles such as latex particles, gel particles, and industrial particles. The substance A contained in the sample flow F1 is not limited to a single substance, and a plurality of different substances can be simultaneously contained to detect a plurality of different interactions.

  The substance B is not particularly limited as long as it is a substance that can flow through the flow path 1 and can interact with the substance A. For example, the substance B is a fluorescent substance such as a fluorescent dye, an antibody, a radioactive substance, a micro substance, and the like. Examples include various reagents such as beads, pH adjusting solution, concentration adjusting solution, hemolyzed blood, and physiological saline, prepared solutions, and chemical solutions. Examples of fluorescent substances include Cascade Blue, Fluorescein isothiocyanate (FITC), Phycoerythrin (PE), Phycoerythrin-Cy5 (PE-Cy5), Phycoerythrin-Cy7 (PE-Cy7), Texas Red, Allophycocyanin (APC), Allophycocyanin-Cy (APC-Cy7) and the like. The substance B contained in the sheath flows F21 and F22 is not limited to a single substance, and a plurality of different substances can be simultaneously contained to detect a plurality of different interactions.

  The substance A and the substance B are allowed to interact with each other in the flow path 1 so that the substance A is chemically or physically modified with the substance B while the substance A and the substance B are allowed to flow through the flow path 1. Can do.

  Further, in this case, as shown by a symbol L3 in FIG. 18, by performing the interaction detection while scanning in the flow direction of the flow path 1, the modification amount of the substance A by the substance B is changed to an arbitrary value in the flow path 1. It can be detected for each position, and the type of substance B and the flow rate of the sheath flow F21 can be controlled by feeding back the modification amount. That is, if the velocity of the fluid in the flow path 1 is set in advance, the reaction time of the substance can be calculated from the position where the interaction is detected.

  Furthermore, if the interaction detection method according to the present invention is used, a method of screening for substance A that interacts with substance B by detecting the presence or absence of interaction in flow channel 1, for example, HIV (Human Immunodeficiency It can be applied to positive reactions of viruses.

  FIG. 19 is a diagram for explaining a state in the flow path 1 when performing the interaction detection method according to the present invention, and shows a laminar flow (sample flow F1, sheath flow F21, F22) different from FIG. It is the cross-sectional schematic diagram seen from the side of the lamination | stacking state.

  In this embodiment, the flow velocity v1 of the sheath flow F21 is set faster than the flow velocity v2 of the sheath flow 22. As described above, by changing the flow rates v1 and v2 of the sheath flow F21 and the sheath flow 22 on both sides of the sample flow F1, the substance A in the sample flow F1 rotates as shown in FIG.

  In this case, for example, by using physiological saline or the like for the sheath flow F22, it is also possible to wash the impurity substance bound to the substance A, the substance B incompletely modified, and the like.

  When the interaction detection method described above is performed in the flow path 1 in which a plurality of laminar flows (sample flow F1, sheath flow F21, F22) are formed as shown in FIGS. 16 and 17, for example, Performing multiple modifications or screening at once by including different types of substances A in the sample flow F1 or different types of substances B in the respective sheath flows F21, F22, F23, F24 Is possible.

<Detection device>
FIG. 20 is a schematic perspective view showing a concept according to an embodiment of the detection device 2 according to the present invention. The detection device 2 according to the present invention roughly includes at least the flow path 1, the first detection unit 31, and the second detection unit 32. Hereinafter, each structure, function, etc. are demonstrated in detail.

(1) Channel 1
The flow channel 1 that can be used in the detection device 2 according to the present invention includes a second detection unit that detects the sample, a flow channel position where the detection is performed by the first detection unit, and the second detection unit. A laminar flow in a state of being stacked perpendicular to the wall surface on the first detection unit side of the flow channel between the position of the flow channel where detection is performed and the wall surface on the second detection unit side. At least a bent portion that is bent so as to be in a parallel laminated state is provided.

  The bent portion provided in the detection unit according to the present invention is configured such that the laminar flow is parallel to the wall surface on the second detection unit side from a state in which the laminar flow is stacked vertically to the wall surface on the first detection unit side. The structure is not particularly limited as long as it can be bent so as to be in a laminated state. In the present embodiment, for example, when the flow direction is the X direction, the flow path is approximately 90 degrees in the Y axis direction. A description will be given below of a bent portion including at least a first bent portion 101 that is bent and a second bent portion 102 that is bent at approximately 90 degrees in the Z-axis direction.

  In this embodiment, the laminating direction of the laminar flow (sample flow F1, sheath flow F21, F22) formed in the flow path 1 is changed by providing the first bent portion 101 and the second bent portion 102 as described above. can do.

For example, as shown in FIG. 20, a laminar flow in a state of being vertically stacked with respect to the wall surface 1d ₁ of the channel 1 facing the detection direction D1 (sample flow F1, sheath flows F21, F22), the first bent by flow through the parts 101 and the second bent portion 102 can be changed to parallel stacked state against the wall surface 1d ₂ of the channel 1 facing the detection direction D2.

  In addition, the 1st bending part 101 and the 2nd bending part 102 are not limited to the form shown in FIG. 20, For example, as shown in FIG. 21, the 1st bending part 101 is about the depth H of the flow path 1. Even if it is this bending, the laminar flow (sample flow F1, sheath flow F21, F22) can be made into the lamination | stacking state parallel to the wall surface 1d2 by letting the 2nd bending part 102 flow after that.

  The channel width, the channel depth, and the channel cross-sectional shape of the channel 1 are not particularly limited as long as they can form a laminar flow, and can be freely designed. For example, the microchannel 1 having a channel width of 1 mm or less can be used in the detection device according to the present invention. In particular, if a microchannel having a channel width of about 10 μm to 1 mm is used, the detection method and the interaction detection method according to the present invention can be more suitably performed.

  As shown in FIG. 22, a sample flow path 11 and a sheath flow path 12 that merges from both sides of the sample flow path 11 are provided upstream of the first bent portion 101 and the second bent portion 102 of the flow path 1. You can also By providing the sample flow channel 11 and the sheath flow channel 12, the sample flow F1 introduced from the sample flow channel 11 is sandwiched from both sides by the sheath flows F21 and F22 introduced from the sheath flow channel 12 The laminar flow can be easily formed.

  Further, as shown in FIG. 22, a sheath flow channel 13 that merges from both sides of the flow channel 1 can be provided downstream of the first bent portion 101 and the second bent portion 102 of the flow channel 1. By providing the sheath flow channel 13, it is possible to form a laminar flow in a state where the sample flow F <b> 1 is converged at the center of the flow channel 1.

In the case of the detection apparatus 2 shown in FIG. 22, the first bent portion 101 and the second bent portion 102 are not limited to the form shown in FIG. 22. For example, as shown in FIG. 23, the first bent portion 101 is used. Even if it is a bend of about the depth H of the flow path 1, a laminar flow (sample flow F 1, sheath flow F 21, F 22) is made to flow on the wall surface 1 d ₂ by passing the second bent portion 102 thereafter. On the other hand, it can be made into the lamination | stacking state parallel to it.

  The flow path 1 is not limited to the form of FIGS. 20-23, and can be designed freely. For example, as shown in FIG. 24, a flow path 1 formed on a two-dimensional or three-dimensional substrate T such as plastic or glass can be used in the detection apparatus according to the present invention. Furthermore, as shown in FIG. 25, a plurality of flow paths 1 can be combined and used.

  Reference numeral 111 denotes a sample flow inlet, and reference numerals 121 and 131 denote sheath flow inlets, respectively. The sheath flow inlets 121 and 131 are individually provided in the sheath flow passages 12 and 13, respectively, but are not particularly limited to this configuration. For example, the sheath flow introduction ports 121a and 131a and the sheath flow introduction ports 121b and 131b (see FIG. 26 (i)) of the respective sheath flow channels 12 and 13 are connected to FIGS. 26 (ii), (iii), and (iv). ), The number of the sheath flow introduction ports 121 and 131 on the substrate T can be reduced, and the working efficiency of the sheath flow introduction can be increased. It becomes possible.

(2) First detection unit 31
In the first detection unit 31, the laminar flow width of the laminar flow (sample flow F <b> 1, sheath flow F <b> 21, F <b> 22) and the position in the flow channel 1 at least upstream of the first bent portion 101 and the second bent portion 102 of the flow channel 1. Is detected.

  If the laminar flow (sample flow F1, sheath flow F21, F22) and the position in the flow path 1 can be detected in the first detection unit, the detection method is not particularly limited, and a known method is freely adopted. be able to. For example, an optical method, an electrical method, a magnetic method, etc. can be mentioned. In particular, in the present invention, an optical method is suitable.

  Examples of optical methods include fluorescence measurement, scattered light measurement, transmitted light measurement, reflected light measurement, diffracted light measurement, ultraviolet spectroscopic measurement, infrared spectroscopic measurement, Raman spectroscopic measurement, FRET measurement, FISH measurement, and various other spectrum measurements. The method is mentioned. Further, if detection is performed using an area imaging device such as a charge coupled device (CCD) or a complementary metal oxide semiconductor (CMOS), an image emitted from the sample C on the entire screen can be photoelectrically converted.

  The type of light irradiation when the optical method is employed is not particularly limited, but light having a constant light direction, wavelength, and light intensity is desirable in order to reliably generate fluorescence and scattered light from the sample. As an example, a laser or LED (Light Emission Diode) can be mentioned.

  In addition, the light irradiation method in the case of adopting the optical method is not particularly limited. For example, as shown by the symbol W1 in FIG. 20, even if the optical information is detected while scanning the irradiation spot in the channel width direction, Good.

  As an electrical method, for example, a resistance value, a capacitance value (capacitance value), an inductance value, an impedance, a change value of an electric field between electrodes, and the like related to a substance such as the sample C contained in the sample flow F1 and the sheath flows F21 and F22. The method of measuring is mentioned.

  Examples of the magnetic method include a method in which a magnetic material is modified on the surface of a substance such as the sample C contained in the sample flow F1 or the sheath flows F21 and F22 to measure magnetization, magnetic field change, magnetic field change, and the like. It is done.

  The number of the first detection units 31 provided in the detection device 2 according to the present invention is not particularly limited. As shown in FIG. 25, when a plurality of flow paths 1 are provided, a plurality of first detection units 31 are also provided. Is also free.

  In the case of the embodiment shown in FIG. 25, the laminar flow width of the laminar flow (sample flow F1, sheath flow F21, F22) in the plurality of flow paths 1 by scanning one first detection unit 31 and It is also possible to detect the position in the flow path 1.

(2) Second detection unit 32
The second detector 32 detects information emitted from the sample C at least downstream of the first bent portion 101 and the second bent portion 102 of the flow path 1. Note that the detection method can be freely selected as in the first detection unit 31.

  The detection condition in the second detection unit 32 can be controlled based on the laminar flow information obtained by the first detection unit 31. For example, the position of the light irradiation spot, the shape of the irradiation spot, the light irradiation intensity, the pulse width, the pulse ratio, the focal position, the amplification factor of the photodetector, the amplification factor of the light irradiation device, and the like can be controlled.

  The number of the second detection units 32 provided in the detection device 2 according to the present invention is not particularly limited, and a plurality of second detection units 32 may be provided. For example, although not shown, when a difference in reaction amount such as interaction in the flow channel 1 is to be detected for each position of the flow channel 1, a plurality of second detection units 32 are provided in the flow direction of the flow channel 1. It can also be provided.

  In this case, it is also possible to detect a difference in reaction amount for each position of the flow path 1 by scanning one second detection unit 32.

  As described above, the detection device 2 according to the present invention includes the first detection unit 31 that detects laminar flow information in the flow channel 1, separately from the second detection unit 32 that actually detects the sample C. Therefore, laminar flow information can be detected while the sample 1 is allowed to flow through the flow channel 1, and this information can be used for detection condition control of the sample C by the second detection unit 32. Therefore, it is not necessary to perform a preparatory process for distributing reference spectrum beads or reference diameter beads corresponding to the sample C in advance, and it is possible to reduce detection time and improve detection accuracy.

  In addition, since the detection device 2 according to the present invention includes the flow path 1 including the first bent portion 101 and the second bent portion 102, after the laminar flow information is detected by the first detector 31, The laminating direction of the laminar flow (sample flow F1, sheath flow F21, F22) can be changed. Therefore, the detection from the first detection unit 31 and the second detection unit 32 can be performed from the same direction. Therefore, it is possible to reduce the size of the apparatus.

It is a flowchart of the detection method which concerns on this invention. It is a perspective schematic diagram of the flow path 1 which can be used for the detection method which concerns on this invention. It is a perspective schematic diagram of the flow path 1 different from FIG. 2 which can be used for the detection method which concerns on this invention. FIG. 4A is a diagram for comparing the inside of the flow channel 1 before and after the laminar flow bending step (II), and FIG. 4A is a schematic cross-sectional view showing the flow channel 1 viewed from the detection direction D, and FIG. These are the cross-sectional schematic diagrams which show the flow path 1 seen from the side with respect to the detection direction D, and FIG. 4 (i-1) and (ii-1) are before performing a laminar flow bending process (II), respectively. FIG. 4 (i-2) and (ii-2) are cross-sectional schematic diagrams showing the channel 1 after performing the laminar flow bending step (II), respectively. . It is a perspective schematic diagram of the flow path 1 different from FIG.2 and FIG.3 which can be used for the detection method which concerns on this invention. FIG. 6 is a schematic perspective view of a flow channel 1 different from those in FIGS. 2, 3, and 5 that can be used in the detection method according to the present invention. It is a perspective schematic diagram of the flow path 1 different from FIG.2, FIG.3, FIG.5 and FIG.6 which can be used for the detection method which concerns on this invention. It is a perspective schematic diagram of the flow path 1 different from FIG.2, FIG.3 and FIG.5-7 which can be used for the detection method which concerns on this invention. FIG. 9 (i) is a schematic cross-sectional view of the laminar flow (sample flow F1, sheath flow F21, F22) after undergoing the laminar flow information detection step (IV) as viewed from the side of the laminated state. Before performing the control step (V), FIG. 9 (ii) is a schematic sectional view showing the state after the laminar flow control step (V). FIG. 10 is a schematic cross-sectional view of a laminar flow (sample flow F1, sheath flow F21, F22) different from that in FIG. 9 after undergoing the laminar flow information detection step (IV), as viewed from the side in a laminated state. ) Is a schematic cross-sectional view before the laminar flow control step (V) is performed, and FIG. 10 (ii) is a schematic cross-sectional view after the laminar flow control step (V) is performed. FIG. 9 is a schematic cross-sectional view of a laminar flow (sample flow F1, sheath flow F21, F22) different from FIG. 9 and FIG. FIG. 11 (i) is a schematic cross-sectional view before the laminar flow control step (V) is performed, and FIG. 11 (ii) is a schematic cross-sectional view after the laminar flow control step (V) is performed. FIG. 12 is a schematic cross-sectional view of a laminar flow (sample flow F1, sheath flow F21, F22) different from that in FIGS. 9 to 11 after the laminar flow information detection step (IV) is viewed from the side of the laminated state. 12 (i) is a schematic cross-sectional view before the laminar flow control step (V) is performed, and FIG. 12 (ii) is a schematic cross-sectional view after the laminar flow control step (V) is performed. FIG. 13 (i) is a schematic cross-sectional view of the laminar flow (sample flow F1, sheath flow F21, F22) after passing through the laminar flow information detection step (IV) as viewed from the side of the laminated state. FIG. 12 (ii) is a schematic cross-sectional view before the control step (VI) is performed and after the detection condition control step (VI) is performed. It is a perspective schematic diagram of the flow path 1 different from FIG.2, FIG.3, FIG.5 to FIG. 8 which can be used for the detection method which concerns on this invention. It is a perspective schematic diagram of the flow path 1 different from FIG.2, FIG.3, FIG.5 to FIG.8, and FIG. 14 which can be used for the detection method which concerns on this invention. FIG. 16 (i) is a top schematic plan view of the flow path 1 formed on the substrate T that can be used in the detection method according to the present invention, and FIG. 16 (ii) is a symbol in FIG. 16 (i). It is a cross-sectional schematic diagram of the flow path 1 of the broken-line circle part shown by R1. FIG. 17 (i) is a schematic cross-sectional view of the flow path 1 of the broken-line circle portion indicated by reference numeral R2 in FIG. 16 (i) as viewed from the detection direction D2, and FIG. 17 (ii) is the same as FIG. It is the cross-sectional schematic diagram which looked at the flow path 1 of the broken-line circle | round | yen part shown by middle code | symbol R2 from the flow direction. It is a figure for demonstrating the mode in the flow path 1 at the time of performing the interaction detection method based on this invention, and the cross section which looked at the laminar flow (sample flow F1, sheath flow F21, F22) from the side of the lamination | stacking state FIG. It is a figure for demonstrating the mode in the flow path 1 at the time of performing the interaction detection method which concerns on this invention, and is laminar flow (sample flow F1, sheath flow F21, F22) different from FIG. FIG. FIG. 2 is a schematic perspective view showing a concept according to an embodiment of a detection device 2 according to the present invention. FIG. 21 is a schematic perspective view showing a concept according to an embodiment different from FIG. 20 of the detection apparatus 2 according to the present invention. It is a perspective model which shows the concept which concerns on embodiment different from FIG.20 and FIG.21 of the detection apparatus 2 which concerns on this invention. It is a perspective model which shows the concept which concerns on embodiment different from FIGS. 20-22 of the detection apparatus 2 which concerns on this invention. It is a perspective model which shows the concept which concerns on embodiment different from FIG.20 and FIG.22 of the detection apparatus 2 concerning this invention. FIG. 25 is a schematic perspective view showing a concept according to an embodiment different from FIGS. 20 to 24 of the detection apparatus 2 according to the present invention. It is a top view schematic diagram which shows the structural example of the flow path 1 which can be used for the detection apparatus 2 which concerns on this invention.

Explanation of symbols

1 passage D1, D2 detection direction F1 sample flow F21, F22, F23, F24 sheath flows _1d 1, 1d ₂ channel wall 11 samples diversion channel 12, 13 sheath diversion channel C sample f1 sample flow width f21, f22 sheath Flow width T Substrate 111 Sample flow inlet 121, 131 Sheath flow inlet A Material A
B Substance B
2 Detection Device 101 First Bent Unit 102 Second Bent Unit 31 First Detection Unit 32 Second Detection Unit

Claims (21)

A method for detecting a sample flowing in a flow path,
A first laminar flow forming step of forming a laminar flow in a state of being stacked perpendicular to the wall surface of the flow channel facing the detection direction by sandwiching a sample flow containing the sample with a sheath flow;
After passing through the first laminar flow forming step, the laminar flow bending step of bending the laminar flow so as to be in a laminated state parallel to the wall surface;
A sample detection step of detecting the sample after the laminar flow bending step;
At least do a detection method.   The laminar flow information detecting step of detecting the laminar flow width and / or position of the laminar flow formed in the first laminar flow forming step is performed after the first laminar flow forming step. The detection method according to 1.   3. The detection method according to claim 2, wherein in the sample detection step, the sample is detected based on the laminar flow information detected in the laminar flow information detection step. The detection method according to any one of claims 1 to 3, wherein at least the following step (1) and step (2) are performed in the laminar flow folding step.
(1) A first bending step of bending the laminar flow approximately 90 degrees in the Y-axis direction, where the laminar flow direction is the X direction.
(2) A second bending step of bending the laminar flow approximately 90 degrees in the Z-axis direction.   5. The detection method according to claim 1, wherein a laminar flow control step of controlling a laminar flow width and / or position of the laminar flow is performed before performing the sample detection step.   The detection method according to any one of claims 1 to 5, wherein a detection condition control step for controlling detection conditions in the sample detection step is performed before the sample detection step.   7. The second laminar flow forming step of further sandwiching the laminar flow with a sheath flow from both sides of the laminated state after the laminar flow bending step is performed. The detection method described.   The detection method according to claim 7, wherein a third laminar flow forming step of forming a plurality of laminar flows is performed by combining two or more laminar flows that have undergone the second laminar flow forming step.   The detection method according to any one of claims 2 to 8, wherein in the laminar flow information detection step, optical detection is performed.   The detection method according to any one of claims 1 to 9, wherein in the sample detection step, optical detection is performed. A method for detecting an interaction between a substance A and a substance B flowing through a flow path,
A sample flow containing the substance A is sandwiched by a sheath flow containing at least a part of the substance B, thereby forming a laminar flow in a state of being stacked perpendicular to the wall surface of the flow channel facing the detection direction. A first laminar flow forming step;
After passing through the first laminar flow forming step, the laminar flow bending step of bending the laminar flow so as to be in a laminated state parallel to the wall surface;
An interaction detecting step for detecting an interaction between the substance A and the substance B after the laminar flow bending process;
At least an interaction detection method.   The laminar flow information detecting step of detecting the laminar flow width and / or position of the laminar flow formed in the first laminar flow forming step is performed after the first laminar flow forming step. 11. The interaction detection method according to 11.   13. The interaction according to claim 12, wherein in the interaction detection step, an interaction between the substance A and the substance B is detected based on the laminar flow information detected in the laminar flow information detection step. Detection method. The interaction detection method according to any one of claims 11 to 13, wherein in the laminar flow bending step, at least the following step (1) and step (2) are performed.
(1) A first bending step of bending the laminar flow approximately 90 degrees in the Y-axis direction, where the laminar flow direction is the X direction.
(2) A second bending step of bending the laminar flow approximately 90 degrees in the Z-axis direction.   The interaction according to any one of claims 11 to 14, wherein a laminar flow control step of controlling a laminar flow width and / or position of the laminar flow is performed before performing the interaction detection step. Detection method.   The interaction detection method according to any one of claims 11 to 15, wherein a detection condition control step for controlling a detection condition in the interaction detection step is performed before the interaction detection step is performed.   17. The second laminar flow forming step of further sandwiching the laminar flow with a sheath flow from both sides of the laminated state after the laminar flow bending step is performed. 18. The interaction detection method as described.   18. The interaction detection method according to claim 17, wherein a third laminar flow forming step of forming a plurality of laminar flows is performed by combining two or more laminar flows that have undergone the second laminar flow forming step.   The detection method according to any one of claims 12 to 18, wherein in the laminar flow information detection step, optical detection is performed.   The interaction detection method according to claim 11, wherein optical detection is performed in the interaction detection step. A detection device for a sample flowing in a flow path,
When the flow direction is the X direction, a first bent portion where the flow path bends approximately 90 degrees in the Y-axis direction, a second bent portion where the flow path bends approximately 90 degrees in the Z-axis direction, A flow path comprising at least
A first detector for detecting a laminar flow width and / or position of a laminar flow containing the sample in at least an upstream flow path of the first bent portion and the second bent portion;
A second detector for detecting the sample in a flow path at least downstream of the first bent portion and the second bent portion;
A detection device comprising at least.

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