Microchips, particle analyzers and measuring devices

Abstract

To reduce the difficulty of optical axis adjustment, to provide a microchip capable of downsizing a whole device, and to provide a particle analysis device, and a measurement device.SOLUTION: A microchip 1 has housings 12A and 12B with a passage 13 through which a sample liquid flows. The housing 12A and 12B are placed so that the sample liquid is illuminated by an excitation optical unit 11 in a light irradiation area on the passage 13. The excitation optical unit 11 has a lens 111. A range of arrangement directions of a plurality of light sources 110a, 110b, 110c in which an emission spot of respective light beams emitted from the plurality of light sources 110a, 110b, 110c is located is within a size of the arrangement direction of the beam of light emitted from the light source 110b immediately after it passes through the lens 111. The light irradiation area is a region where the respective beams intersect after passing through the lens 111.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a microchip, a particle analysis device, and a measurement device. [Background technology]

[0002] As a conventional technique, a flow cytometer has been proposed as a measuring device that detects multiple fluorescence emissions generated when cell particles as a sample labeled with multiple fluorescent dyes are excited by multiple laser light sources without setting a delay time (see, for example, Patent Document 1).

[0003] The flow cytometer as a measuring device disclosed in Patent Document 1 has a plurality of light sources that irradiate a sample flowing through a flow path with a plurality of excitation lights having different wavelengths at a predetermined period and at different phases, and a light-guiding member that guides the plurality of excitation lights onto the same incident light path and focuses the light on dyed particles. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Japanese Patent Application Laid-Open No. 2007-046947 [Patent Document 2] Japanese Patent Application Laid-Open No. 2016-57309 Summary of the Invention [Problem to be solved by the invention]

[0005] Meanwhile, a disposable flow cytometer has been proposed in which the flow cell that forms the flow path through which the sample flows is made into a microchip (see, for example, Patent Document 2).

[0006] The flow cytometer as a measuring device disclosed in Patent Document 2 comprises a flow cell having a flow path formed in a flat substrate, a light irradiation means for irradiating light onto particles in a sample liquid flowing through the flow path, a detection means for detecting scattered light or fluorescence emitted from the particles when light is irradiated, identifying the particles based on the signal intensity, and detecting the target particles, a constant pressure pump for applying pressure pulses to the particles in the sample liquid flowing through the flow path of the flow cell and an electromagnetic valve connected to the pump, and a control means for controlling the operation of the electromagnetic valve based on a signal from the detection means.

[0007] The flow cytometer of Patent Document 1, mentioned above, guides multiple laser beams along the same optical path, detecting forward and side scattered light and fluorescence from cell particles labeled with multiple fluorescent dyes without setting a delay time, simplifying the adjustment of the optical axis from the light source to the flow cell. However, optical axis adjustment of the optical system is still required. In particular, in a configuration that uses multiple excitation wavelengths to measure corresponding emission characteristics, the optical axis must be adjusted for each excitation wavelength light source, which presents a problem that the optical axis adjustment is not easy. In other words, Patent Document 1 does not disclose a specific optical system, and an optical system that does not require or that easily requires optical axis adjustment is desired. Furthermore, the flow cytometer of Patent Document 2 uses a disposable microchip, so the optical axis must be adjusted every time the microchip is replaced, again presenting a problem that the optical axis adjustment is not easy.

[0008] An object of the present invention is to provide a microchip, a particle analysis device and a measurement device that reduce the difficulty of adjusting the optical axis and make the entire device compact. [Means for solving the problem]

[0009] In order to achieve the above object, one aspect of the present invention provides the following microchip and particle analysis device: and providing measurement equipment.

[0010] [1] A microchip having a housing having a flow path through which a sample liquid flows, the housing being arranged so that the sample liquid is irradiated with light by a light irradiation means in a light irradiation region on the flow path, The light irradiation means has a focusing member, and the range in the arrangement direction of the multiple light sources in which the emission spots of each light beam emitted from the multiple light sources exist is within the size in the arrangement direction of the light beam immediately after the light beam emitted from the emission spot located most centrally with respect to the focusing member of the multiple light sources passes through the focusing member, and the light irradiation area is a microchip that is an area where the beams of the multiple light sources intersect after passing through the focusing member. [2] The microchip according to [1], wherein the light irradiated in the light irradiation area of ​​the housing is irradiated via a mirror provided between the light-collecting member of the light irradiation means and the light irradiation area. [3] The microchip according to [2], wherein the mirror rotates so as to change the angle with respect to the light beam, and moves the light irradiation area in accordance with the flow of the sample in the sample solution. [4] The microchip according to [2] or [3], wherein the light irradiation means sequentially irradiates light beams of a plurality of wavelengths and / or a plurality of intensities. [5] The microchip according to [3] or [4], wherein the mirror returns to the initial position where it started tracking when detection of transmitted light, reaction luminescence, fluorescence, or scattered light from the sample in the sample solution in the light irradiation area is discontinued. [6] A particle analyzer for analyzing particles contained in a sample liquid, a microchip having a housing having a flow path through which the sample liquid flows, and a sample liquid reservoir formed on the housing and through which the sample liquid flows in the flow path; a light irradiation means for irradiating the sample liquid flowing through the flow channel in a light irradiation region; a detection means for detecting a characteristic amount of light emitted from the particle; an analyzing means for identifying and analyzing the target particles based on the detection signal of the detecting means, and outputting the results; The light irradiation means has a plurality of light sources and a focusing member, and the range in the arrangement direction of the plurality of light sources in which the emission spots of each light beam emitted from the plurality of light sources exist is within the size in the arrangement direction of the beam immediately after the light beam emitted from the emission spot located most centrally with respect to the focusing member of the plurality of light sources passes through the focusing member, and the light irradiation area is the area where the beams of the plurality of light sources intersect after passing through the focusing member. [7] A microchip having a housing having a flow path through which a sample liquid flows, the housing being arranged so that the sample liquid is irradiated with light by a light irradiation means in a light irradiation region on the flow path, the light irradiation means having a light condensing member, a range in an arrangement direction of the plurality of light sources in which an emission spot of each of the light beams emitted from the plurality of light sources exists is within a size in the arrangement direction of the light beam immediately after the light beam emitted from the emission spot located most centrally with respect to the light condensing member of the plurality of light sources passes through the light condensing member, and the light irradiation region is a region where the beams of the plurality of light sources intersect after passing through the light condensing member; a spectroscope for separating light incident from the sample liquid; and analyzing means for analyzing chemical changes occurring in the sample in the sample liquid by detecting a characteristic amount of the light dispersed by the spectrometer. [Effects of the Invention]

[0011] According to the inventions of claims 1, 6 and 7, the difficulty of adjusting the optical axis is reduced, and the entire device can be made compact. According to the invention of claim 2, a mirror can be provided between the light collecting member and the light irradiation area. According to the invention of claim 3, the light irradiation area can be moved in accordance with the flow of the sample in the sample liquid. According to the invention of claim 4, light beams of a plurality of wavelengths or a plurality of intensities can be emitted sequentially. According to the invention of claim 5, the timing for returning to the initial position where tracking started can be set to the timing when detection of transmitted light, reaction luminescence, fluorescence or scattered light from the sample in the light irradiation area stops. [Brief explanation of the drawings]

[0012] [Figure 1] FIG. 1 is a schematic diagram showing the configuration of a flow cytometer according to the first embodiment. [Figure 2] FIG. 2 is a block diagram showing an example of a process for processing a signal output from a detector. [Figure 3] FIG. 3 is a schematic perspective view showing an example of the configuration of the microchip according to the first embodiment. [Figure 4] 4(a) and (b) are diagrams showing the configuration of an excitation optical unit according to an embodiment of the present invention, with FIG. 4(a) being a plan view and FIG. 4(b) being a side view. [Figure 5] 5(a) and (b) are diagrams showing the configuration of an excitation optical unit according to an embodiment of the present invention, with FIG. 5(a) being a plan view and FIG. 5(b) being a side view. [Figure 6] FIG. 6 shows a simulation of the relationship between the amount of lateral movement of the light source and the oblateness of the cross-sectional shape of the light beam. [Figure 7] 7A and 7B are explanatory diagrams of what happens when the light source is misaligned, where FIG. 7A is a plan view of the light source when it is misaligned, FIG. 7B is a side view, and FIG. 7C is an explanatory diagram of the change in cross-sectional shape. [Figure 8] FIG. 8 is a partial cross-sectional view taken along the line BB' showing an example of the positional relationship between the microchip and the excitation optical unit. [Figure 9] 9(a) and (b) are schematic diagrams showing the relationship between the light irradiation position and the sample flow. [Figure 10] FIG. 10 is a conceptual diagram showing the configuration of a light beam emitting device according to the first modification of the present invention. [Figure 11] FIG. 11 is a conceptual diagram showing the configuration of an excitation optical unit according to the third modification of the present invention. [Figure 12]FIG. 12 is a timing chart illustrating the light source control of the excitation optical unit and the operation control of the MEMS mirror device in the third modification. [Figure 13] 13(a) to 13(c) are schematic diagrams showing an example of the configuration of a cell sorter using a flow cell, which is a microchip according to the second embodiment. [Figure 14] 14(a) to 14(c) are schematic diagrams showing an example of the configuration of a measurement system using a microchip having a housing through which a sample flows according to the third embodiment. DETAILED DESCRIPTION OF THE INVENTION

[0013] [First embodiment] (Flow cytometer configuration) Fig. 1 is a schematic diagram showing the configuration of a flow cytometer according to a first embodiment, and Fig. 2 is a block diagram showing an example of a process for processing signals output from a detector.

[0014] The flow cytometer 2 comprises a hydrodynamic flow mechanism for arranging samples such as cell particles stained with fluorescent labeling reagents in the sample liquid in a line within a sheath flow, an optical mechanism for irradiating each cell particle with multiple lights of different wavelengths and receiving scattered light and fluorescence, and a signal processing unit for controlling and processing electrical signals related to the scattered light and fluorescence output from the optical mechanism.

[0015] The hydrodynamic flow mechanism has a sample suspension supply unit 211 for storing and supplying a sample suspension containing cell particles to be analyzed as a sample liquid, a sheath liquid supply unit 212 for storing and supplying a sheath liquid, a flow chamber 222, and a microchip 1 which is a flow cell connected downstream of the flow chamber 222.

[0016] Flow chamber 222 has a substantially cylindrical shape, and suspension supply pipe 221 is disposed along its central axis. The sample suspension and sheath liquid stored in sample suspension supply unit 211 and sheath liquid supply unit 212 are supplied into suspension supply pipe 221 and flow chamber 222 by the pressure of air pumps 201, 202, respectively. This forms a sheath-shaped sheath flow (laminar sheath flow f2) in which the sheath liquid envelops the sample suspension in a cylindrical shape. At this time, by setting the pressure of sample suspension supply unit 211 slightly lower than the pressure of sheath liquid supply unit 212, hydrodynamic narrowing occurs, and the flow diameter of the sample suspension enveloping the sheath liquid becomes extremely narrow (sample flow f1), allowing cell particles contained in the sample suspension to be aligned in a row within microchip 1.

[0017] The microchip 1 is configured, for example, to be removable and replaceable from the flow chamber 222. It has an orifice at its bottom, from which a sheath flow containing cell particles that have passed through the microchip 1 is jetted. When the jetted sheath flow is vibrated by a vibration device, such as a piezoelectric element (not shown), droplets containing the respective cell particles are formed. Furthermore, when sorting droplets containing specific cell particles (when applied to a cell sorter), a charging unit (not shown) is provided to charge the sheath flow containing the cell particles to be sorted just before the break-off point where the droplets are formed. Furthermore, a pair of deflection plates 23a and 23b, to which a predetermined voltage (e.g., a DC voltage of 6000 V) is applied, are provided. The charged droplets are deflected by a force from a DC electric field as they pass between the deflection plates 23a and 23b, and are then sorted.

[0018] The optical mechanism as the light irradiation means has light sources 110a, 110b, and 110c that irradiate a sample such as cell particles aligned in the microchip 1 with a plurality of excitation lights of different wavelengths, a lens 111 as a light-collecting member, and a mirror (113, see FIG. 11) as needed. The light sources 110a, 110b, and 110c and the lens 111 (and, as a variant, a MEMS mirror device 113) are mounted on a single substrate (112) as shown in FIG. 4, which will be described later, to form a unit as the excitation optical unit 11.

[0019] The light sources 110a, 110b, and 110c may be any light source and are not intended to limit the present invention, but single-mode semiconductor lasers are preferred in that they collimate or focus the light beams. Alternatively, the light sources 110a, 110b, and 110c may be directly irradiated onto the lens 111, or the light sources 110a, 110b, and 110c may be disposed at any position and light guided to the lens 111 using an optical fiber or the like. The number of light sources 110a, 110b, and 110c is not limited to three as long as they satisfy the arrangement conditions described below.

[0020] 1, the light emitted from the light sources 110a, 110b, and 110c is irradiated onto the microchip 1 via a lens 111. Details of the light sources 110a, 110b, and 110c, the lens 111, and the light irradiation position (light irradiation area L on the microchip 1) determined by their arrangement and their relative positional relationship will be described in detail later.

[0021] The optical mechanism includes two detectors for detecting light: a detector 14A that detects forward-scattered light (FSC) scattered by the sample, and a detector 14B that detects side-scattered light (SSC) scattered by the sample and multiple fluorescent lights (FL) of various wavelengths generated by exciting the sample. Detector 14A has a bandpass filter (not shown, e.g., a selected wavelength of 488±5 nm) that selectively transmits light of a specific wavelength, and detects forward-scattered light scattered by the sample. Detector 14B, on the other hand, detects side-scattered light having three wavelengths identical to the wavelengths of the irradiated light and fluorescent light having various wavelengths different from the wavelengths of the irradiated light from each light source. Detector 14B has filters, such as a dichroic long-pass filter, a long-pass filter, a short-pass filter, and a band-pass filter, corresponding to each detection wavelength, as well as multiple detectors corresponding to each detection wavelength.

[0022] As shown in FIG. 2 , the signal processing unit includes a signal processing unit 15, a data analysis unit 16, and a light source control unit 17. The output of the detector 14A (14B) is input to the signal processing unit 15. The signal processing unit 15 is configured using known technologies, such as an A / D conversion circuit that converts analog signals to digital signals, an AWH calculation circuit that calculates the area, width, and height of the signal, a parameter selector circuit that selects the optical signal elements to be processed, and a compensation circuit that compensates for fluorescence leakage. The signal processing unit 15 outputs the analog signal detected by the detector 14A (14B) as a numerical value (digital signal) of the wavelength and intensity of light. The data analysis unit 16 analyzes multiple light intensity distributions and their interrelationships, and outputs the results to the light source control unit 17. The light source control unit 17 controls the light source based on the analysis results of the data analysis unit 16 (described in detail in Modification 3). The data analysis unit 16 analyzes data from a known flow cytometer, calculates ratios and correlations between each light intensity, analyzes the characteristics of the sample, and outputs the results. Furthermore, the characteristic quantity of light is not limited to the intensity of each wavelength, but may be a comparative value of the intensity of each wavelength or a physical quantity calculated from the comparative value of the intensity. If a spectroscope is provided, wavelength distributions may be compared.

[0023] (Microchip configuration) FIG. 3 is a schematic perspective view showing an example of the configuration of the microchip according to the first embodiment.

[0024] The microchip 1 has housings 12A and 12B that form a flow path 13 therein. A sheath liquid, such as physiological saline, flows within the flow path 13 as a sheath flow to surround the sample flow, forming a sheath flow f2 that flows around the periphery and a sample flow f1 that is formed in the center of the sheath flow for introducing the sample. As shown in FIG. 1, the microchip 1 also has an inlet path upstream of each flow and an outlet path downstream, but these are omitted in FIG. 3. An excitation optical unit 11 is disposed on the incident surface of the housing 12B, and irradiates the sample in the sample flow f1 flowing through the housing with excitation light via the excitation optical unit 11. Here, the area on the sample flow f1 of the microchip 1 that should be irradiated with light is referred to as the "light irradiation region L." As will be described below, the area where light from multiple light sources intersects among the light irradiated by the excitation optical unit 11 is referred to as the "light irradiation position."

[0025] The following description will be given of a case in which the excitation optical unit 11 is configured integrally with the microchip 1 so as to eliminate the need to adjust the light irradiation area on the microchip 1, but the microchip 1 and the excitation optical unit 11 may have a fitting structure and be fitted into each other so that the light irradiation position of the excitation optical unit 11 coincides with the light irradiation area L on the sample flow f1 of the microchip 1, or the microchip 1 and the excitation optical unit 11 may be installed using an alignment attachment or the like so that the light irradiation position of the excitation optical unit 11 coincides with the light irradiation area L on the sample flow f1 of the microchip 1. Furthermore, as long as the excitation optical unit 11 is positioned so that the light irradiation position of the excitation optical unit 11 coincides with the light irradiation area L on the sample flow f1 of the microchip 1, the excitation optical unit 11 may not be formed as a unit, and the light sources 110a to 110c and the lens 111 may be prepared separately. Furthermore, the microchip 1 and the excitation optical unit 11 may be arranged at different positions and by any method as long as they are arranged so that the light irradiation position of the excitation optical unit 11 coincides with the light irradiation region L on the sample flow f1 of the microchip 1. These arrangement methods can be changed as appropriate depending on the design, such as making the microchip 1 and the excitation optical unit 11 disposable, making only the microchip 1 disposable, or making the microchip 1 and the lens 111 disposable.

[0026] Within the channel 13, laminar flows of sample flow f1 and sheath flow f2 are formed from top to bottom in the drawing (see also FIG. 8), with the sample flow f1 being held at the center of the sheath flow f2. While the cross-sectional shape of the channel 13 is rectangular in FIG. 3, it may also be circular. The sample flow f1 has a diameter of, for example, 10 to 100 μm, and the sheath flow f2 has a diameter of 100 to 1000 μm. When the channel cross-section is polygonal, these diameters can be considered to be the diameters of the inscribed circle. For example, the sample flow f1 contains a sample with a diameter of approximately 10 μm (any size may be selected within the range of 0.5 to 50 μm), and the sample is a microparticle, such as a cell or lymphocyte.

[0027] The sample flow f1 and the side-scattered light / fluorescence light path ls continuing to the detector 14B are all positioned on one identical plane (A-A' cross section), while the optical path of the illumination light from the excitation optical unit 11, the sample flow f1, and the forward-scattered light path lf continuing to the detector 14A are all positioned on another identical plane (B-B' cross section). The one identical plane and the other identical plane are preferably positioned at an angle appropriate for the optical signal to be detected, for example, orthogonal to each other. The housings 12A and 12B are formed by a method described below and are made of materials such as silicon, glass, and resin that do not interfere with the excitation light or light from the sample. The housings 12A and 12B transmit at least the excitation light, forward-scattered light, and side-scattered light / fluorescence emitted from the excitation optical unit 11. The A-A' cross section is the cross section of the plane where the sample flow f1 is positioned when viewing the light irradiation surface of the microchip 1 from the excitation optical unit 11 side in FIG. 1. The BB' cross section is a cross section of the plane where the sample flow f1 is located when the microchip 1 is viewed from the front from the detector 14B side.

[0028] (Configuration of excitation optical unit) Figures 4(a) and (b) and Figures 5(a) and (b) are diagrams showing the configuration of an excitation optical unit according to an embodiment of the present invention, with Figures 4(a) and 5(a) being plan views and Figures 4(b) and 5(b) being side views. Note that although the figures show three light sources, the number of light sources can be any number as long as it is two or more.

[0029] The excitation optical unit 11 is composed of a plurality of light sources 110a to 110c and a lens 111, which is a single condenser member. In this case, a condenser lens is used as the lens 111. The light sources 110a to 110c are arranged at intervals x. The light source 110b and the lens 111 are arranged at a distance y1. The emission spots e of the light sources 110a to 110c a ~e c The light beam r emitted from a ~r c is the light trail c of the central axis of light a ~c c so that the light beams r are parallel to each other.a ~r c The shape of the light beam spreads as it progresses toward the lens 111. Note that the term "single light collecting element" here means that the light collecting element is shared by multiple light sources, and the light collecting element does not necessarily have to be made up of a single lens, and may be made up of a compound lens.

[0030] After that, the light is collected by the lens 111, and after being collected by the lens 111, the light beams are collimated and are reflected by the light traces c of the central axes of the light emitted from the light sources 110a to 110c. a ~c c The collimated light beam is a light beam that travels without changing its diameter. The light trace c of the central axis of this light a ~c c The point where these light beams intersect will be called the convergence point (light irradiation area L). The distance y2 between this convergence point and the condenser lens serving as lens 111 does not need to be mathematically exact and equal to the focal length of lens 111. Furthermore, the distance between light sources 110a to 110c and lens 111 and the design of lens 111 will be adjusted so that the beam diameter of the area where the three light beams intersect at the convergence point (light irradiation area L) is, for example, about 100 μm, in accordance with the size of the sample (about 10 μm).

[0031] Here, the distance D between the light sources 110a to 110c in the arrangement direction is set to 1 / 200 of the light beam r from the light source 110a arranged at the center. a The light beam r immediately after passing through the lens 111 a The size of the light beam r in this case is set to be within the range of Q. a The diameter of the beam is calculated by approximating the optical beam to a Gaussian beam and dividing the optical power by 1 / e from the value on the central axis. 2 (e: Napier's number) is the diameter of the light beam r a The light beam r immediately after passing through the lens 111 a The size Q is the beam diameter immediately after passing through the lens if the lens 111 is a lens, and is the beam diameter immediately after being reflected by the reflector if the lens 111 is made of a reflector.

[0032] The light beams r of the plurality of light sources 110a to 110c a ~r c Even if the beam diameters immediately after passing through the lens 111 are different, the beam diameter of the light source 110a arranged at the most center is used.

[0033] The reason why the above configuration makes it possible to irradiate the light beams of the plurality of light sources onto the light irradiation position will be explained below.

[0034] Figure 6 shows a simulation of the relationship between the lateral movement of the light source and the oblateness of the cross-sectional shape of the light beam. Here, the simulation was performed with a collimated beam diameter (corresponding to Q in Figure 5) of 1 mm, a focal length of the single lens of 6 mm, and an aperture of the single lens of 4 mm. As can be seen from Figure 3, when the lateral movement is around 0.5 mm, the oblateness exceeds 10%, and when it exceeds 0.5 mm, the oblateness increases rapidly. Normally, it is desirable to use an oblateness within one digit to avoid degrading beam quality. Note that the oblateness is expressed as flattening = 1 - ellipticity, and the ellipticity is expressed as the ratio of the minor axis to the major axis of the ellipse.

[0035] This result shows that when the light source 110a is placed beyond the range Q (range of -0.5 mm to 0.5 mm) in FIG. 5, the light beam r a This indicates that the shape of the light source 110a is significantly deteriorated. Furthermore, this condition generally holds true under practical conditions (the radiation angle of the light source 110a is within 30 degrees and the collimated beam diameter is within 5 mm), regardless of the type of focusing lens, the diameter of the light beam, the focal length of the lens, or the radiation angle of the light source 110a, and even if the radiation angle of the light source 110a after horizontal movement differs from the radiation angle of the light source 110a at its original position. In other words, since the diameter of the collimated beam is Q, the horizontal axis in FIG. 6 can be interpreted as "lateral movement of the light source (mm)" instead of "lateral movement of the light source / Q" (dimensionless). Therefore, it is sufficient if the excitation optical unit 11 is configured so that "lateral movement of the light source / Q" ≦ 1 is satisfied.

[0036] From this reinterpretation of the relationship "lateral movement of light source / Q," the position of the light source, the position of the lens, and the focal length of the lens can be designed. For example, in FIG. 5, consider a case where two light sources 110a and 110b (110b and 110c) are arranged with a spacing x of 0.15 mm, the distance y2 from optical lens 111 to the light irradiation position (light irradiation area L) is set to 11.5 mm, and the light beam diameter is narrowed to 100 μm. First, when a light beam is emitted from light source 100b to optical lens 111, the divergence angle of the beam from the semiconductor laser light source is usually about 10°, so the distance y1 between light source 110a and optical lens 111 is about 11.3 mm, the value of Q is 1 mm, and the value of "lateral movement of light source / Q" is 0.15. That is, the range in the arrangement direction of the multiple light sources 110a, 110b, and 110c is within the size in the arrangement direction of the light beam emitted from the emission spot (light source 110b) located most centrally with respect to the light collecting member (optical lens 111) immediately after passing through the light collecting member (optical lens 111). Furthermore, by using optical lens 111 with a focal length of 5.7 mm, for example, a light beam with a diameter Q = 1 mm can be focused to 100 μm at the light irradiation position (light irradiation area L) when the distance y2 from the optical lens 111 to the light irradiation position (light irradiation area L) is 11.5 mm.

[0037] The distance y2 between the optical lens 111 and the light irradiation position, ie, 11.5 mm, given as an example above, is determined based on the following conditions. (a1) Assume that the sample flow f1 has a diameter of 100 μm, the surrounding sheath flow f2 has a diameter of 1 mm (an inscribed circle), and the sheath flow f2 is closed (surrounded by the housings 12A and 12B (cell chips)). A cell wall (e.g., 1 mm thick) is located outside the sheath flow f2. Furthermore, if the exit port of the excitation optical unit 11 is located 10 mm from the outside of the sheath flow f2 cell wall and an optical lens 111 is disposed at the exit port, the distance between the optical lens 111 and the sample is "half the diameter of the sheath flow f2 (0.5 mm)" + "1 mm thickness of the cell wall" + "10 mm distance from the outside of the sheath flow f2 cell wall to the exit port of the excitation optical unit 11," and therefore the distance between the optical lens 111 and the light irradiation position is 11.5 mm.

[0038] In addition, in the following examples, the positions of the light sources, the positions of the lenses, and the focal lengths of the lenses can be designed in a similar manner. (a2) In the same arrangement as (a1) above, if the optical lens 111 is offset from the exit (end) of the excitation optical unit 11 (for example, an offset of 5 mm), the distance y2 between the optical lens 111 and the light irradiation position becomes 16.5 mm by adding the offset length (11.5 + 5). The space created by the offset is used, for example, for arranging a MEMS mirror (113, FIG. 11) described later, etc.

[0039] In this case, when two light sources 110a and 110b (110b and 110c) are arranged with a spacing x of 0.15 mm and the light beam diameter is narrowed to 100 μm at the light irradiation position (light irradiation area L) y2 = 16.5 mm away from the optical lens 111, the distance x1 between the light source 110a and the optical lens 111 is approximately 8.7 mm, and the value of Q is 1 mm, so the value of "lateral movement amount of light source / Q" is 0.15 as above. Furthermore, by using the optical lens 111 with a focal length (for example, 5.7 mm), the light beam with Q = 1 mm can be narrowed to 100 μm at the light irradiation position (light irradiation area L) y2 = 10.5 mm away from the optical lens 111.

[0040] (b1) Furthermore, in the case where the sheath flow f2 is open (i.e., not surrounded by the housings 12A and 12B (cell chip)), there is no cell wall outside the sheath flow f2, so the exit port of the excitation optical unit 11 is located 10 mm from the sheath flow f2. If an optical lens is placed at this exit port, the distance between the optical lens 111 and the sample is "half the diameter of the sheath flow f2 (0.5 mm)" plus "the distance from the outside of the sheath flow f2 cell wall to the exit port of the excitation optical unit 11, 10 mm," so the distance y2 between the optical lens 111 and the light irradiation position is 10.5 mm.

[0041] In this case, when two light sources 110a and 110b (110b and 110c) are arranged with a spacing x of 0.15 mm and the light beam diameter is narrowed to 100 μm at the light irradiation position (light irradiation area L) y2 = 10.5 mm away from the optical lens 111, the distance y1 between the light source 110a and the optical lens 111 is approximately 12.5 mm, and the value of Q is 1 mm, so the value of "lateral movement amount of light source / Q" is 0.15. Furthermore, by using the optical lens 111 with a focal length (for example, 5.7 mm), the light beam with Q = 1 mm can be narrowed to 100 μm at the light irradiation position (light irradiation area L) y2 = 10.5 mm away from the optical lens 111.

[0042] Furthermore, even if the distance y2 between the optical lens 111 and the sample is smaller than the above, this can be accommodated by changing the focal length of the optical lens 111. For example, when the distance between the optical lens 111 and the sample is y2=1.5 mm and an optical lens 111 with a focal length of 1.71 mm is used, the distance y1 between the light source 110a and the optical lens 111 may be set to approximately 7.1 mm. When a light beam is input from the light source 100b to the optical lens 111, the divergence angle of the beam from the semiconductor laser light source is usually about 10°. Therefore, when the distance y1 between the light source 110a and the optical lens 111 is approximately 7.1 mm, the value of Q is 0.3 mm, and the value of "lateral movement amount of the light source / Q" is 1 or less. In other words, the range of the arrangement direction of the multiple light sources 110a, 110b, and 110c is within the size of the arrangement direction of the light beam emitted from the emission spot (light source 110b) located most centrally with respect to the focusing member (optical lens 111) immediately after passing through the focusing member (optical lens 111). Furthermore, by using the optical lens 111 with a focal length of 1.71 mm, a light beam with a diameter Q = 0.3 mm can be focused to 100 μm at the light irradiation position (light irradiation area L) at a distance y2 = 1.5 mm from the optical lens 111 to the light irradiation position (light irradiation area L). Similarly, for example, when the distance y2 between the optical lens 111 and the sample is 6.5 mm, an optical lens 111 with a focal length of 1.71 mm can be used, and the distance y1 between the light source 110a and the optical lens 111 can be set to approximately 2 mm. In this way, various arrangements are possible by changing each optical parameter. In addition, such an example in which the distance between the optical lens 111 and the sample is relatively short is assumed to be a case in which the microchip 1 and the excitation optical unit 11 are configured as an integrated unit, or a case in which the excitation optical unit 11 is configured as an insertable unit into the microchip 1.

[0043] In addition, the radiation angle is not isotropic, and the light beam r emitted from the light source 110a a Even if the cross-sectional shape of the light source is originally an ellipse, the same can be said if the beam diameter in the direction of the light source arrangement (horizontal direction) is used. From this, it can be said that the range D in the direction of the arrangement of the light sources in which the emission spots of each light beam emitted from multiple light sources exist is the light beam r aIt can be seen that it is desirable that the diameter Q of the light beam in the arrangement direction of the light sources immediately after passing through the lens 111.

[0044] 7A and 7B are explanatory diagrams showing the case where the light source is misaligned, in which FIG. 7A is a plan view showing the case where the light source is misaligned, FIG. 7B is a side view, and FIG. 7C is an explanatory diagram showing the change in the cross-sectional shape. As shown in the figure, the light beam r emitted from the light source 110a a is collimated by the lens 111 (a single lens in the figure), and when the light source 110a is moved in the horizontal direction (the direction in which the light sources are arranged: the vertical direction in the figure), the light beam r a is deformed as shown in Figure 7(a). In particular, the light beam r a The cross-sectional shape of the light beam r a When the light beam r is emitted isotropically, it is distorted from a circle to an ellipse as shown in Figure 7(c). a The lens 111 may be designed so that the cross section thereof has a circular shape.

[0045] FIG. 8 is a partial cross-sectional view taken along the line BB′ showing an example of the positional relationship between the microchip 1 and the excitation optical unit 11. As shown in FIG.

[0046] By appropriately setting the performance of the light sources 110a-110c and the lens 111 and their relative positions, the excitation optical unit 11 irradiates a light irradiation area L on the sample flow f1 with light from the light sources 110a-110c, each emitting light of a different wavelength, from a position at a distance d from the irradiation surface of the opposing housing 12B. The irradiation range of the light irradiation area L is determined depending on the distance d, the thickness t of the housing 12B, the distance from the housing 12B to the sample flow f1, and the refractive indices of air, the material of the housing 12B, the sample flow f1, and the sheath flow f2. The irradiation range of the light irradiation area L is designed to be approximately 100 μm, for example, to match the size of the sample. However, a lens may be provided on the housing 12B or the like to focus a beam of a smaller or larger diameter.

[0047] 9(a) and (b) are schematic diagrams showing the relationship between the light irradiation region L and the sample flow f1.

[0048] As shown in FIG. 9(a), when the sample flow f1 is set upside down in a plan view, the light trace c of the central axis of the light a ~c c The position where these light sources intersect with each other is set as a light irradiation area L, and by passing through this light irradiation area L, light from all of the light sources 110a to 110c can be efficiently emitted.

[0049] Furthermore, as shown in FIG. 9(b), when the sample flow f1 is set upside down in front view, the light trace c of the central axis of the light a ~c c The position where these light sources intersect with each other is set as a light irradiation area L, and by passing through this light irradiation area L, light from all of the light sources 110a to 110c can be efficiently emitted.

[0050] (Effects of the first embodiment) According to the above-described embodiment, an excitation optical unit is provided integrally with the microchip 1 or separately so that it can be aligned with the light irradiation area, and multiple light beams from multiple light sources are configured to overlap in the light irradiation area L by the lens 111, and the light irradiation area L is designed to be at a position where the sample flow f1 of the microchip 1 intersects. Therefore, there is no need to adjust the optical axes of the multiple light sources each time the microchip is replaced, and further, because the excitation optical unit 11 is used, the entire device can be made more compact than when an optical system is used in which the optical axes of multiple light sources must be aligned with each other.

[0051] (Variation 1) 10 is a conceptual diagram of a light beam emitting device according to Modification 1 of the present invention. Modification 1 is the same as the first embodiment except that the number of light sources is six.

[0052] Two red semiconductor lasers 110r1 and 110r2 with an oscillation wavelength of 635 nm, two blue semiconductor lasers 110b1 and 110b2 with an oscillation wavelength of 450 nm, and two green semiconductor lasers 110g1 and 110g2 with an oscillation wavelength of 520 nm are arranged in parallel at an interval of 0.15 mm. In this case, too, the range D in the element arrangement direction of the emission spot between the blue semiconductor laser 110b1 and the green semiconductor laser 110g2 arranged at the extreme ends is within the beam range Q (the lateral range of the light beam from the red semiconductor laser 110r1: 1 mm) immediately after passing through the condenser lens 24 of the red semiconductor laser 110r1 arranged closest to the center of the condenser lens 24. The arrangement may be, for example, blue, red, green, blue, red, green, or any other order from the condenser lens 24 side.

[0053] In addition to the advantages of the first embodiment, the first modification of the present invention simplifies the arrangement of six semiconductor lasers, enabling a light beam with up to twice the intensity of the first embodiment to be obtained, thereby realizing a high-brightness light beam projection device. Furthermore, all six semiconductor lasers may be the same color, or any number of the six semiconductor lasers may be used. In other words, the intensity of the irradiated light can be set by the number of elements. As a result, in the conventional case where the intensity of the irradiated light is changed, changing the supply current value changes the light emission state (e.g., the light emission position and beam diameter of the light beam), requiring readjustment of the optical axis, etc. However, in this modification, the intensity can be set by the number of elements without changing the state of each light source, resulting in a stable optical system and eliminating the need for optical readjustment.

[0054] (Variation 2) Next, a light beam emitting device according to a second modification of the present invention will be described. The second modification is the same as the first embodiment except that the number of light sources is increased to five by adding infrared and ultraviolet light sources. Note that the diagram is omitted because it is similar to FIG. 10 except for the number of light sources.

[0055] A red semiconductor laser with an oscillation wavelength of 635 nm, a blue semiconductor laser with an oscillation wavelength of 450 nm, a green semiconductor laser with an oscillation wavelength of 520 nm, an infrared semiconductor laser with an oscillation wavelength of 830 nm, and an ultraviolet semiconductor laser with an oscillation wavelength of 375 nm are arranged in parallel in this order at intervals of 0.15 mm. In this case, too, the range D in the element arrangement direction of the emission spot of the red semiconductor laser placed at the extreme end and the emission spot between the ultraviolet semiconductor lasers is set to fall within the beam range Q of the green semiconductor laser immediately after passing through the condenser lens. Note that the arrangement may be such that any semiconductor laser is placed at the center of the condenser lens, and any order is acceptable.

[0056] In the second modification of the present invention, in addition to the effects of the first embodiment, by using five semiconductor lasers with different oscillation wavelengths, it is possible to realize a compact light beam projection device consisting of multiple light beams in a wider wavelength range, from infrared light to ultraviolet light, compared to the first embodiment.

[0057] (Variation 3) (Mirror tracking control) 11 is a conceptual configuration diagram of an excitation optical unit according to Modification 3 of the present invention. In Modification 3, a MEMS mirror device 113 for changing the optical axis direction at a predetermined angle is provided between the light sources 110a to 110c and the light irradiation area L of the excitation optical unit according to the first embodiment. Specifically, the center 113ac of the mirror surface of the movable mirror portion 113a of the MEMS mirror device 113 is positioned with respect to the center of the optical axis, so that the reflected light can be scanned.

[0058] The size of the MEMS mirror device 113 is a chip size of 7 mm x 5 mm x 0.7 mm (length x width x height). The size of the movable mirror portion 113a of the MEMS mirror device 113 is 1 mmφ. The MEMS mirror device 113 uses Si as the base material, and an Al film is used as the material of the movable mirror portion 113a.

[0059] For example, by driving this movable mirror portion 113a by a piezoelectric drive method with a maximum drive voltage of ±15V, it is possible to use a device capable of performing two-dimensional optical scanning with a high-speed (horizontal) axis drive frequency of 35 kHz, a high-speed (horizontal) axis oscillation angle of ±15 degrees (mirror oscillation angle), a low-speed (vertical) axis drive frequency of 60 Hz, and a low-speed (vertical) axis oscillation angle of ±15 degrees (mirror oscillation angle) (in this modification example 3, one-dimensional scanning is performed using a MEMS mirror capable of two-dimensional optical scanning.) Here, a piezoelectric drive method using the piezoelectric effect is used, but a drive device of an electrostatic drive method or an electromagnetic drive method may also be used.

[0060] The excitation optical unit of Modification 3 of the present invention has a function of changing the optical axis, and this change can be made to coincide with the flow direction of the sample. Therefore, under control of the light source control unit 17, light can be irradiated to follow the flow of the sample, namely, the sample flow f1. Hereinafter, light source control and operation control of the MEMS mirror device for achieving effective tracking will be described. Note that, although one-dimensional optical scanning in the sample flow direction will be described here, two-dimensional scanning including not only the flow direction of the sample but also its cross-sectional direction may be performed depending on the configuration of the microchip 1.

[0061] FIG. 12 is a timing chart illustrating the light source control of the excitation optical unit and the operation control of the MEMS mirror device in the third modification.

[0062] First, for example, the light source control unit 17 controls the excitation optical unit 11 to irradiate the sample flow f1 with red excitation light (1). At this point, the mirror rotation angle of the movable mirror portion 113a of the MEMS mirror device 113 is set to a position that irradiates the upstream limit position of the sample flow f1, and this is set as the initial position. If a sample is present in the sample flow f1, scattered light corresponding to the red excitation light is observed by the detector 14A (2). Furthermore, if the detector is able to track the sample flow, scattered light is continuously observed, and the rotational angular velocity of the movable mirror portion 113a is configured to be automatically or manually adjusted to ensure continuous observation.

[0063] Next, when scattered light begins to be observed as in (2) above, the light source control unit 17 moves the mirror rotation angle of the movable mirror unit 113a of the MEMS mirror device 113 downstream over time in accordance with the flow of the sample flow f1 at the adjusted rotation angle (3). The red excitation light continues to be irradiated to confirm that the mirror rotation angle is tracking the sample, but irradiation may be stopped when irradiating the green excitation light, which will be described next.

[0064] In the above state, it is confirmed that the excitation light reflected by the mirror is tracking the sample, and the light source control unit 17 then controls the excitation optical unit 11 to further irradiate light of a wavelength different from the red excitation light, for example, green excitation light (4). If a sample is present in the sample flow f1, fluorescence corresponding to the green excitation light is observed by the detector 14B (5). The irradiation of the green excitation light and the observation of the fluorescence are carried out for a predetermined period. The predetermined period may be determined from the cycle determined by (1) to (8).

[0065] Next, when the irradiation of the green excitation light is completed, the light source control unit 17 controls the excitation optical unit 11 to further irradiate, for example, blue excitation light (6). If a sample is present in the sample flow f1, fluorescence corresponding to the blue excitation light is observed by the detector 14B (7). The irradiation of the blue excitation light and the observation of the fluorescence are carried out for a predetermined period. The predetermined period may also be determined from the cycle determined by (1) to (8).

[0066] Next, when scattered light in response to the red excitation light is no longer observed, the light source control unit 17 resets the mirror rotation angle of the movable mirror unit 113a of the MEMS mirror device 113 to the initial position (8) and moves on to tracking the next sample. By repeating these operation cycles from (1) to (8) for the next sample that flows by, the scattered light and fluorescence are tracked to the sample and observed.

[0067] Although an example of sequential irradiation with excitation light of different wavelengths has been shown, excitation light of the same wavelength may be repeatedly irradiated sequentially, or may be irradiated sequentially with varying intensity. Furthermore, in addition to simultaneous irradiation with multiple wavelengths, multiple wavelengths may be irradiated sequentially so as to increase the excitation level in order. Furthermore, light may be irradiated using a combination of these.

[0068] In the third modification of the present invention, the sample is irradiated with light of multiple wavelengths or multiple intensities, or a combination of these, so that more information can be obtained from a single sample. Furthermore, it is possible to obtain the generation of multi-stage excited states and the corresponding reactions, which are not possible with irradiation of a single light or irradiation of light for a limited period of time.

[0069] [Second embodiment] In the second embodiment, the excitation optical unit 11 of the first embodiment is applied to a disposable chip-type flow cell, which is an example of a microchip of the present invention, with the wavelength appropriately changed, to form a particle separation device (cell sorter) equipped with a particle analysis device using the chip-type flow cell.

[0070] 13(a) to 13(c) are schematic diagrams showing an example of the configuration of a cell sorter using a flow cell, which is a microchip according to the second embodiment.

[0071] The cell sorter 2B includes a flow cell 1B having a flat substrate as a housing and a flow path formed within the flat substrate, an excitation optical unit 11B as a light irradiation means for irradiating light onto particles in the sample liquid flowing through the flow path, a detector 730 as a detection means for detecting scattered light, reaction light, or fluorescence of at least a specific wavelength generated from the particles when irradiated with light, a microcomputer 790 as an analysis means for identifying particles and distinguishing target particles based on the signal intensity of these detection signals, a constant pressure pump that applies pressure to particles in the sample liquid flowing through the flow path of the flow cell 1B, and an electromagnetic valve connected to it, and the microcomputer 790 controls the operation of the electromagnetic valve based on the analysis results. The excitation optical unit 11B is configured to be integrated with the flow cell 1B or to be attachable without requiring position adjustment of the light irradiation area.

[0072] Figures 13(a) and (b) show the disposable chip-type flow cell 1B connected to a cylinder pump and an electromagnetic valve. Figure 13(b) shows the CC cross section of Figure 13(a). Flow cell 1B has a structure in which separation channels 4-1 and 4-2, and separation reservoirs 5-1 and 5-2, respectively, are symmetrically added. The disposable chip-type flow cell is made of transparent resin. Resins that can be used include polymetal acrylate (PMMA), cycloolefin copolymer (COC), and methylpentene polymer. Methylpentene polymer is particularly suitable for use with a laser in the UV wavelength range (approximately 350 to 410 nm) as the irradiation light source. Wavelengths that can be selected include 405 nm, 488 nm, 561 nm, and 637 nm. In this case, the wavelengths of the light sources of excitation optical unit 11B can be any combination of three wavelengths, for example, 405 nm, 488 nm, 561 nm, and 637 nm. The fluorescence wavelength range can be 445 nm center wavelength for 405 nm, 543 nm center wavelength for 488 nm, 591.5 nm center wavelength for 561 nm, and 676 nm, 716 nm, or 775 nm center wavelength for 637 nm. A separation reservoir is also installed in flow path 4-2, creating a symmetrical structure, and an electromagnetic valve and a cylinder pump are connected.

[0073] Figure 13(c) shows the connection relationship between the optical system and the control circuit. The cross-sectional view of the chip is the DD cross-section of Figure 13(a). As shown in Figure 13(a), air pressure is applied to the upstream side of the chip to flow the sample liquid and sheath liquid through the channel 370. Light of a single or multiple wavelengths is irradiated from the excitation optical unit 11B onto the center of the main channel, slightly upstream of the opposing separation channels 4-1 and 4-2. The moment the target particle cells in the sample liquid pass through this irradiated area, scattered light and fluorescent light are generated in pulses. The scattered light is separated into the same wavelength as the irradiated light by a spectrometer 10B composed of multiple filters and detected by a detector 730. The fluorescent light, which has a longer wavelength than the irradiated light, is separated by the spectrometer 10B into multiple wavelength regions and each is detected by the detector 730. Each detection pulse signal is amplified, AD converted, and digitized in a signal processing unit 740, and the characteristics of the cells in the sample liquid are analyzed by a microcomputer 790 serving as a data analysis unit. This completes the operation of the particle analyzer.

[0074] Furthermore, when used as a separation device, the data analysis unit further determines whether the analysis results satisfy preset separation conditions, and if so, outputs a trigger signal to the electromagnetic valve driver 760 after a certain delay. This delay is adjusted to the time it takes for cells to flow from the laser irradiation area to the separation channels 4-1 and 4-2. Upon receiving the trigger signal, the electromagnetic valve driver 760 outputs a signal to open the electromagnetic valve 7-1 or 7-2 for a certain period of time. When the electromagnetic valve is open, the target cells flow into the separation reservoir 5-1 or 5-2. The sheath fluid and sample fluid that do not flow into the separation reservoir flow into the collection reservoir 210.

[0075] As shown in Figure 13(a), a constant air pressure is applied to the reservoir 300 upstream of the chip by a constant-pressure pump (not shown). Inside the reservoir 300 is the sample fluid reservoir 200, which contains sample fluid containing cells appropriately marked with a fluorescent reagent or the like, and sheath fluid outside of it. Phosphate buffer saline (PBS) is preferably used as the sheath fluid. A common air pressure causes the sample fluid 20 to flow downstream via the sample fluid pipe 20-1, and the sheath fluid flows downstream via the left and right sheath fluid pipes 30-1 and 30-2. After the three pipe channels merge, the sample fluid flows through the channel 370, narrowed by the sheath fluid. For example, the width of the channel 370 can be 80 μm and the depth can be 50 μm. The width of the sample fluid after merging is approximately 1 / 10 of the channel width. The flow channel 370 has separation channels 4-1 and 4-2 that face each other from the side, and these channels are connected to reservoirs 5-1 and 5-2. A central portion of the flow channel 370, located a few hundred micrometers upstream of this facing region, is irradiated with light that combines light with wavelengths of, for example, 405 nm, 488 nm, and 561 nm. The beam size is an ellipse with a length of 50 μm and a width of 20 μm.

[0076] (Effects of the second embodiment) According to the second embodiment described above, the cell sorter and the microchip used in the cell sorter employ an excitation optical unit 11 similar to that of the first embodiment, so the difficulty of adjusting the optical axis each time the microchip used in the cell sorter is replaced is reduced or eliminated, and the entire device can be made more compact than when an optical system is employed in which the optical axes of multiple light sources must be aligned as excitation light sources.

[0077] [Third embodiment] In the third embodiment, the excitation optical unit 11 of the first embodiment is applied to a measurement system using a microchip.

[0078] 14(a) to 14(c) are schematic diagrams showing an example of the configuration of a measurement system using a microchip having a housing through which a sample flows according to the third embodiment, where Fig. 14(a) is a plan view, Fig. 14(b) is a front cross-sectional view in the case where transmitted light or scattered light is detected, and Fig. 14(c) is a plan cross-sectional view in the case where scattered light, fluorescence, or reaction luminescence is detected.

[0079] Measurement system 3 includes pump 4, pump 5, microchip 1C having flow path 31 into which sample liquid A flows from pump 4, flow path 32 into which sample liquid B flows from pump 5, and flow path 33 where these flow paths converge, recovery unit 6 for the converged liquid, excitation optical unit 11C that irradiates light of multiple wavelengths onto the converged sample liquid in microchip 1C, spectrometer 10C formed in microchip 1C that disperses light incident from the sample using multiple filters, detector 14 that detects light features corresponding to the irradiated light and detects the light dispersed by spectrometer 10C as analysis means for analyzing chemical changes, signal processing unit 15, and data analysis unit 16. The wavelengths used in the waveguide multiplexer and waveguide spectrometer satisfy the above-mentioned conditions for the multiplexer and spectrometer and can be selected from the excitation spectrum, absorption spectrum, emission spectrum, and fluorescence spectrum; for example, wavelengths selected from the peaks of the spectra can be used.

[0080] An example of operation is explained below. Sample solution A, to which a protonated dye and an ionophore such as crown ether that selectively complexes with specific ions has been added, is flowed inside microchip 1C. When a target ion is present in the merged sample solution B, the ion undergoes a complexation reaction with the ionophore in sample solution A. This shift in the absorption spectrum of the dye in sample solution A causes a change in the intensity of detection light of a specific wavelength that passes through sample solution A. Based on the above principle, the presence or absence of target ions can be measured as an example of analyzing chemical changes that have occurred in a sample.

[0081] 14(b), excitation optical unit 11C irradiates the dye in sample solution A with light of a wavelength selected corresponding to the absorption spectrum (before and after the shift) of the dye in sample solution A, or with light of a wavelength corresponding to the absorption spectrum (before, after, or both) of multiple dyes, combined together. Spectrometer 10C separates the light irradiated by excitation optical unit 11C into specific wavelengths corresponding to the light, and detects the light intensity as an example of a feature of the light passing through sample solution A using detector 14, signal processing unit 15, and data analysis unit 16, and determines the change in the light intensity. Spectrometer 10C is positioned so that it can detect the light irradiated by excitation optical unit 11C and transmitted through sample solution A.

[0082] 11(c), the spectrometer 10C may be configured to irradiate the dye in the sample solution A with excitation light and receive scattered light, fluorescence, or reaction luminescence in a direction perpendicular to the direction of irradiation of the light, as in the first and second embodiments. The spectrometer 10C then separates the light into wavelengths corresponding to the light irradiated by the excitation optical unit 11C, and the detector 14, signal processing unit 15, and data analysis unit 16 detect the characteristic quantities of the light emitted from the dye in the sample solution A.

[0083] (Effects of the third embodiment) According to the third embodiment described above, the excitation optical unit 11 is provided integrally with the microchip 1C or without the need for optical axis adjustment. Therefore, even when detecting chemical changes in a flow path in a microchip into which multiple phases flow, the difficulty of adjusting the optical axis each time the microchip is replaced is reduced or eliminated, and the entire device can be made more compact than when an optical system is used that requires the optical axes of multiple light sources to be aligned.

[0084] [Other embodiments] The present invention is not limited to the above-described embodiment, and various modifications can be made without departing from the spirit of the present invention. Various modifications are possible.

[0085] In the above embodiment, the excitation optical unit is formed integrally with the microchip, but if alignment is possible, the excitation optical unit may be attached to the microchip during manufacturing or preparation for measurement, or may be attached using an attachment or the like that connects them.

[0086] Furthermore, as a detector, any known detector that can detect weak light can be used, such as an APD (Avalanche Photodiode), and examples of array detectors include an APD array and a multi-channel PMT (PhotoMultiplier Tube). [Explanation of symbols]

[0087] 1: Microchip 1B: Flow cell 1C: Microchip 2: Flow cytometer 2B: Cell sorter 3: Measurement system 4, 5: Pump 6: Collection section 10B, 10C: Spectrometer 11, 11B, 11C: Excitation optical unit 12A, 12B: Housing 13: Flow path 14, 14A, 14B: Detector 15: Signal processing section 16: Data analysis section 17: Light source control unit 20: Sample solution 24: Condenser lens 31, 32, 33: Flow path 110: Light source 111: Lens 112: Substrate 113: MEMS mirror device

Claims

1. A light irradiation means having a plurality of light sources and a focusing member, wherein the range in the arrangement direction of the plurality of light sources in which the emission spots of the respective light beams emitted from the plurality of light sources exist is within the size in the arrangement direction of the light beam immediately after the light beam emitted from the emission spot located most centrally with respect to the focusing member of the plurality of light sources has passed through the focusing member, and the light irradiation area is an area where the beams of the plurality of light sources intersect after passing through the focusing member, wherein the light beams emitted from the emission spots located most centrally with respect to the focusing member of the plurality of light sources are light beams that expand in the direction in which light is emitted, and the beams immediately after passing through the focusing member are light beams whose width of the luminous flux of the light beam does not change in the direction in which light is emitted or light beams that converge in the direction in which light is emitted; A microchip comprising: a housing having a flow path through which a sample liquid flows, the housing being arranged so that the light irradiation region is positioned above the flow path.

2. 2. The microchip according to claim 1, wherein the light irradiated in the light irradiation region of the housing is irradiated via a mirror provided between the light condensing member of the light irradiating means and the light irradiation region.

3. 3. The microchip according to claim 2, wherein the mirror rotates so as to change an angle with respect to the light beam, and moves the light irradiation area in accordance with the flow of the sample in the sample solution.

4. 4. The microchip according to claim 2, wherein said light irradiating means sequentially irradiates light beams of a plurality of wavelengths and / or a plurality of intensities.

5. 5. The microchip according to claim 3, wherein the mirror returns to an initial position at which it starts tracking when detection of transmitted light, reaction luminescence, fluorescence, or scattered light from the sample in the sample solution in the light irradiation area is discontinued.

6. A microchip as described in claim 1, wherein the microchip and the light irradiation means have an interlocking structure so that the light irradiation position of the light irradiation means coincides with the light irradiation area on the flow path of the microchip, and the light irradiation means is installed on the microchip by this interlocking structure.

7. A microchip as described in claim 1, further having an attachment for alignment so that the light irradiation position of the light irradiation means coincides with the light irradiation area on the flow path of the microchip, and the light irradiation means is installed on the microchip by the attachment.

8. A particle analyzer for analyzing particles contained in a sample liquid, comprising: a microchip having a housing having a flow path through which the sample liquid flows, and a sample liquid reservoir formed on the housing and through which the sample liquid flows in the flow path; a light irradiation means for irradiating the sample liquid flowing through the flow channel in a light irradiation region; a detection means for detecting a characteristic amount of light emitted from the particle; an analyzing means for identifying and analyzing the target particles based on the detection signal of the detecting means, and outputting the results; The light irradiation means has a plurality of light sources and a focusing member, and the range in the arrangement direction of the plurality of light sources in which the emission spots of each light beam emitted from the plurality of light sources exist is within the size in the arrangement direction of the beam immediately after the light beam emitted from the emission spot located most centrally with respect to the focusing member of the plurality of light sources passes through the focusing member, and the light irradiation area is the area where the beams of the plurality of light sources intersect after passing through the focusing member, and the light beam emitted from the emission spot located most centrally with respect to the focusing member of the plurality of light sources is a light beam that spreads in the direction of light emission, and the beam immediately after passing through the focusing member is a light beam whose luminous flux width does not change in the direction of light emission or a light beam that converges in the direction of light emission.

9. a microchip having a housing having a flow path through which a sample liquid flows, the housing being arranged so that the sample liquid is irradiated with light by a light irradiation means in a light irradiation region on the flow path, the light irradiation means having a light condensing member, wherein a range in an arrangement direction of the plurality of light sources in which emission spots of the respective light beams emitted from the plurality of light sources exist is within a size in the arrangement direction of the light beam immediately after the light beam emitted from the emission spot located most centrally with respect to the light condensing member of the plurality of light sources has passed through the light condensing member, the light irradiation region is a region where the beams of the plurality of light sources intersect after passing through the light condensing member, the light beam emitted from the emission spot located most centrally with respect to the light condensing member of the plurality of light sources is a light beam that spreads in the light emitting direction, and the beam immediately after passing through the light condensing member is a light beam that does not change in width of the luminous flux of the light beam in the light emitting direction or a light beam that converges in the light emitting direction; a spectroscope for separating light incident from the sample liquid; and analyzing means for analyzing chemical changes occurring in the sample in the sample liquid by detecting a characteristic amount of the light dispersed by the spectrometer.

Images