JP7780798B2 - Microchips, particle analyzers and measuring devices - Google Patents

Description

The present invention relates to microchips, particle analysis devices, and measurement devices.

As a conventional technique, a flow cytometer has been proposed as a measuring device that detects multiple fluorescent light emissions generated when cell particles, serving 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).

The flow cytometer disclosed in Patent Document 1 as a measuring device has multiple light sources that irradiate a sample flowing through a flow path with multiple excitation light beams having different wavelengths at a predetermined cycle and at different phases, and a light-guiding member that guides the multiple excitation light beams along the same incident light path and focuses them on dyed particles.

Japanese Patent Application Laid-Open No. 2007-046947 JP 2016-57309 A

On the other hand, a disposable flow cytometer has been proposed in which the flow cell that forms the channel through which the sample flows is made into a microchip (see, for example, Patent Document 2).

The flow cytometer disclosed in Patent Document 2 as a measuring device comprises a flow cell having a flow path formed within 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 irradiated with light, identifying the particles based on the signal strength, and detecting the target particles; a constant-pressure pump for applying pressure pulses to the particles in the sample liquid flowing through the flow cell flow path 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.

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. This simplifies the adjustment of the optical axis from the light source to the flow cell, but 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 the light source of each excitation wavelength, which poses a problem: the optical axis adjustment is not easy. In other words, Patent Document 1 does not disclose specific optical systems, and there is a demand for optical systems that do not require or that easily require optical axis adjustment. Furthermore, the flow cytometer of Patent Document 2 uses disposable microchips, so the optical axis must be adjusted each time the microchip is replaced, again posing a problem: the optical axis adjustment is not easy.

The object of the present invention is to provide a microchip, particle analysis device, and measurement device that reduces the difficulty of optical axis adjustment and makes the entire device more compact.

In order to achieve the above object, one aspect of the present invention provides the following microchip, particle analysis device, and measurement device.

[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 condensing member of the light irradiating 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 liquid.
[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, 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.

[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 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 passes through the light condensing member, and the light irradiation region is a region where the respective 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.

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 second aspect of the present invention, a mirror can be provided between the light collecting member and the light irradiation area.
According to the third aspect of the present invention, the light irradiation area can be moved in accordance with the flow of the sample in the sample liquid.
According to the fourth aspect of the present invention, 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 the timing when detection of transmitted light, reaction luminescence, fluorescence or scattered light from the sample in the light irradiation area stops.

FIG. 1 is a schematic diagram showing the configuration of a flow cytometer according to the first embodiment. FIG. 2 is a block diagram showing an example of a process for processing a signal output from a detector. FIG. 3 is a schematic perspective view showing an example of the configuration of the microchip according to the first embodiment. 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. 5(a) and 5(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. 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. 7A and 7B are explanatory diagrams illustrating a case where the light source is misaligned, where FIG. 7A is a plan view of the case where the light source is misaligned, FIG. 7B is a side view, and FIG. 7C is an explanatory diagram illustrating the change in cross-sectional shape. FIG. 8 is a partial cross-sectional view taken along the line B-B' showing an example of the positional relationship between the microchip and the excitation optical unit. 9(a) and (b) are schematic diagrams showing the relationship between the light irradiation position and the sample flow. FIG. 10 is a conceptual diagram showing the configuration of a light beam emitting device according to the first modification of the present invention. FIG. 11 is a conceptual diagram showing the configuration of an excitation optical unit according to a third modification of the present invention. 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 according to the third modification. 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. 14A to 14C 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.

[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.

Flow cytometer 2 includes a hydrodynamic flow mechanism for arranging samples, such as cell particles stained with fluorescently labeled reagents in a sample liquid, in a line within a sheath flow; an optical mechanism for irradiating each cell particle with multiple light beams of different wavelengths and receiving scattered light and fluorescent light; and a signal processing unit for controlling and processing electrical signals related to the scattered light and fluorescent light output from the optical mechanism.

The hydrodynamic flow mechanism has a sample suspension supply unit 21 ₁ for storing and supplying a sample suspension containing cell particles to be analyzed as a sample liquid, a sheath liquid supply unit 21 ₂ for storing and supplying a sheath liquid, a flow chamber 22 ₂ , and a microchip 1 which is a flow cell connected downstream of the flow chamber 22 ₂ .

Flow chamber _22-2 has a substantially cylindrical shape, with suspension supply pipe _22-1 disposed along its central axis. The sample suspension and sheath liquid stored in sample suspension supply unit _21-1 and sheath liquid supply unit _21-2 are supplied into suspension supply pipe _22-1 and flow chamber _22-2 by the pressure of respective air pumps _20-1 , _20-2 . As a result, a sheath-shaped sheath flow (laminar sheath flow f _-2 ) is formed 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 _21-1 slightly lower than the pressure of sheath liquid supply unit _21-2 , hydrodynamic constriction occurs, and the flow diameter of the sample suspension enveloping the sheath liquid becomes extremely narrow (sample flow f _-1 ), allowing cell particles contained in the sample suspension to be aligned in a row within microchip 1.

The microchip 1 is configured, for example, to be removable and replaceable from the flow chamber _22-2 . 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 that charges 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 23 a, 23 b to which a predetermined voltage (e.g., a DC voltage of 6000 V) is applied is provided. The charged droplets are deflected by a force from a DC electric field as they pass between the deflection plates 23 a, 23 b, and are then sorted.

The optical mechanism as the light irradiation means has, for light irradiation, 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-condensing member, and a mirror (113, see FIG. 11) as needed. The light sources 110a, 110b, and 110c and the lens 111 (and, as a modified example, a MEMS mirror device 113) are mounted on a single substrate (112) and unitized as an excitation optical unit 11, as shown in FIG. 4 described below.

Light sources 110a, 110b, and 110c may be any light source and are not intended to limit the present invention. However, single-mode semiconductor lasers are preferred because they collimate or focus the light beams. Furthermore, in addition to irradiating lens 111 directly from light sources 110a, 110b, and 110c, light sources 110a, 110b, and 110c may be positioned at any location and light guided to lens 111 using an optical fiber or the like. Furthermore, the number of light sources 110a, 110b, and 110c is not limited to three, as long as the placement conditions described below are met.

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

The optical mechanism also includes detector 14A, which detects forward-scattered light (FSC) scattered by the sample, and detector 14B, which 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 specific wavelengths and detects forward-scattered light scattered by the sample. Detector 14B, on the other hand, detects side-scattered light of the three wavelengths identical to the wavelengths of the irradiating light and fluorescent light of various wavelengths different from the wavelengths of the irradiating light from each light source. It has filters such as dichroic long-pass filters, long-pass filters, short-pass filters, and band-pass filters corresponding to each detection wavelength, as well as multiple detectors corresponding to each detection wavelength.

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 composed of 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 the intensity distributions of multiple light beams 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 Variation 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 light feature is not limited to the intensity of each wavelength, but may also be a comparison value of the intensity of each wavelength or a physical quantity calculated from the comparison value of the intensity. If a spectroscope is provided, wavelength distributions may also be compared.

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

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 excitation light is irradiated onto the sample in the sample flow _f1 flowing through the housing 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."

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 unitized, 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 whether the microchip 1 and the excitation optical unit 11 are disposable, whether only the microchip 1 is disposable, or whether the microchip 1 and the lens 111 are disposable.

Within the flow 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 flow 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 cross section of the flow channel 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 fine particle, such as a cell or lymphocyte.

Furthermore, 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 the method described below using 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 located 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.

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

The excitation optical unit 11 is composed of a plurality of light sources 110a to 110c and a lens 111, which is a single focusing element. In this case, a focusing 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 light beams r _a to r _c emitted from the emission spots e _a to _ec of the light sources 110a to _{110c travel toward the lens 111 while expanding in shape so that the light traces c a to cc of the central} axes of the light are parallel to one another. Note that the term "single focusing element" used here expresses that the focusing element is shared by a plurality of light sources, and the focusing element does not necessarily have to consist of a single piece and may be composed of a compound lens.

The light is then focused by lens 111. After being focused by lens 111, the light beams travel as collimated beams such that the light traces c _a to c _c of the central axes of the light emitted from each of light sources 110 a to 110 c intersect. A collimated light beam refers to a light beam that travels without changing its beam diameter. The point where the light traces c _a to c _c of the central axes of the light intersect is referred to as the convergence point (light irradiation area L). The distance _y2 between this convergence point and the focusing lens serving as lens 111 does not need to be mathematically exacting and equal to the focal length of lens 111. The distance between light sources 110 a to 110 c and lens 111 and the design of lens 111 are 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, approximately 100 μm, in accordance with the size of the sample (approximately 10 μm).

Here, the spacing D in the arrangement direction of the multiple light sources 110a to 110c is set to be within a range equal to or less than the size Q of the light beam _{r a from} the centrally arranged light source 110a immediately after passing through the lens 111. In this case, the diameter of the light beam r _a is set to be the diameter at which the light beam is approximated to a Gaussian beam and the optical power is 1/e ² (e: Napier's number) of the value on the central axis. Note that the size Q of the light beam _{r a immediately} after passing through the lens 111 is the beam diameter immediately after passing through the lens if the lens 111 is a lens, or is the beam diameter immediately after being reflected by the reflector if the lens 111 is made of a reflector.

Even if the beam diameters of the light beams r _a to r _c from the multiple light sources 110 a to 110 c are different immediately after passing through the lens 111, the beam diameter of the light source 110 a arranged at the center is used.

The following explains why the above configuration allows light beams from multiple light sources to be irradiated onto the light irradiation position.

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 single lens focal length of 6 mm, and a single lens aperture 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. It is generally 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.

This result indicates that if the light source 110a is positioned beyond the range Q (-0.5 mm to 0.5 mm) in Figure 5, the shape of the light beam r _a is significantly degraded. 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 moving laterally 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 Figure 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 for the excitation optical unit 11 to be configured so that "lateral movement of the light source/Q" ≦ 1 is satisfied.

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, consider the case in FIG. 5 where two light sources 110a and 110b (110b and 110c) are arranged with a spacing x of 0.15 mm, the distance _y2 from the 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 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 about 11.3 mm, the value of Q becomes 1 mm, and the value of "lateral movement of light source/Q" becomes 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 condenser member (optical lens 111) immediately after passing through the condenser member (optical lens 111). Furthermore, by using optical lens 111 with a focal length of 5.7 mm, for example, it is possible to focus a light beam with a diameter Q = 1 mm 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.

The distance y ₂ = "11.5 mm" between the optical lens 111 and the light irradiation position given as an example above is determined based on the following conditions.
(a1) When 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 (for example, 1 mm thick) is assumed to exist 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.

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 in (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). Note that the space created by the offset is used, for example, for arranging a MEMS mirror (113, FIG. 11) described later, etc.

In this case, when two light sources 110a and 110b (110b and 110c) are arranged at a distance x = 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 having 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.

(b1) Furthermore, in a case that differs from the above (a1) in that the sheath flow _f2 is open (a case in which the sheath flow _f2 is 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 , and if an optical lens is placed 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)" + "the distance 10 mm from the outside of the sheath flow f2 cell wall to the exit port of the excitation optical unit 11," so the distance _y2 between the optical lens 111 and the light irradiation position is 10.5 mm.

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 having 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.

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 projected from the light source 100b onto 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 becomes 0.3 mm, and the value of "lateral movement amount of the light source/Q" becomes 1 or less. In other words, 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 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.

Furthermore, even if the radiation angle is not isotropic and the cross-sectional shape of the light beam r _a emitted from the light source 110 a is originally elliptical, the same can be said if the beam diameter in the light source arrangement direction (horizontal direction) is used. From this, it can be seen that it is desirable that the range D in the light source arrangement direction, in which the emission spots of the light beams emitted from the multiple light sources exist, be within the range Q of the diameter of the light beam in the light source arrangement direction immediately after the light beam r _a passes through the lens 111.

FIG. 7 is an explanatory diagram of what happens when the light source is misaligned, with FIG. 7( a) being a plan view, FIG. 7( b) being a side view, and FIG. 7( c) being an explanatory diagram of the change in cross-sectional shape. As shown in the figure, when a light beam r _a emitted from a light source 110 a is collimated by a lens 111 (a single lens in the figure), if the light source 110 a is moved in the horizontal direction (the light source arrangement direction: the vertical direction in the figure), the light beam r _a is deformed as shown in FIG. 7( a). In particular, if the light beam r _a is emitted isotropically, the cross-sectional shape of the light beam r _a will be distorted from a circle to an ellipse as shown in FIG. 7( c). Note that the lens 111 may be designed to take this distortion into account so that the cross-sectional shape of the light beam r _a becomes circular.

Figure 8 is a partial cross-sectional view taken along the line B-B' showing an example of the positional relationship between the microchip 1 and the excitation optical unit 11.

The excitation optical unit 11 irradiates a light irradiation area L on the sample flow f1 with light from the light sources 110a to 110c, each emitting light of a different wavelength, from a position at a distance d from the irradiation surface of the opposing housing 12B by setting the performance and positional relationship of the light sources 110a to 110c and the lens ₁₁₁ to appropriate values. Here, 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 . Note that the irradiation range of the light irradiation area L is designed to be, for example, about 100 μm in accordance with the size of the sample, but a lens may be provided on the housing 12B or the like to focus a beam of a smaller or larger diameter.

9(a) and (b) are schematic diagrams showing the relationship between the light irradiation area L and the sample flow _f1 .

As shown in FIG. 9( a), when the sample flow _f1 is set upside down in a plan view, the position where the light traces c _a to c _c of the central axis of the light intersect with each other is set as the light irradiation region L, and by passing through this light irradiation region L, it is possible to efficiently irradiate light from all of the light sources 110 a to 110 c.

Furthermore, as shown in FIG. 9( b), when the sample flow _f1 is set upside down when viewed from the front, the position where the light traces c _a to c _c of the central axis of the light intersect with each other is set as the light irradiation area L, and by passing through this light irradiation area L, it is possible to efficiently irradiate light from all of the light sources 110 a to 110 c.

(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 as to be positionable relative to 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 the 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.

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

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. Note that the arrangement may be in the order of blue, red, green, blue, red, green from the condenser lens 24 side, or any other order.

In Variation 1 of the present invention, in addition to the effects of the first embodiment described above, the arrangement of six semiconductor lasers is simplified, allowing for a light beam with up to twice the intensity compared to the first embodiment, 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 activated. This means that the intensity of the irradiated light can be set by the number of elements. As a result, in the past, when changing the irradiated light intensity, changing the supply current value would change the light emission state (light beam emission position and beam diameter), requiring readjustment of the optical axis, etc. However, in this variation, 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.

(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. The diagram is omitted because it is similar to FIG. 10 except for the number of light sources.

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 sequence 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 focusing lens. Note that any semiconductor laser can be placed at the center of the focusing lens, and any order is acceptable.

In variant 2 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.

(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 reflected light can be scanned.

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 for the movable mirror portion 113a.

For example, by driving this movable mirror portion 113a using a piezoelectric drive system 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 and a high-speed (horizontal) axis oscillation angle of ±15 degrees (mirror oscillation angle), and 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 variant 3, one-dimensional scanning is performed using a MEMS mirror capable of two-dimensional optical scanning). Here, a piezoelectric drive system using the piezoelectric effect is used, but electrostatic or electromagnetic drive systems may also be used.

The excitation optical unit of the third modification 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 the control of the light source control unit 17, light can be irradiated to follow the flow of the sample, i.e., 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.

Figure 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 Modification 3.

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 sample flow can be tracked, scattered light is continuously observed, and the rotation angular velocity of the movable mirror portion 113a is configured to be automatically or manually adjusted to ensure continuous observation.

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 at the adjusted rotation angle in accordance with the flow of the sample flow _f1 (3). The red excitation light continues to be irradiated to confirm that the mirror rotation angle is tracking the sample, but the irradiation may be stopped when irradiating the green excitation light, which will be described next.

In the above state, since it is confirmed that the excitation light reflected by the mirror is tracking the sample, 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).

Next, after completing the irradiation of the green excitation light, 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. This predetermined period may also be determined from the cycle determined by (1) to (8).

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 its initial position (8) and moves on to tracking the next sample. By repeating these operation cycles (1) to (8) for the next sample that flows by, the sample is tracked and scattered light and fluorescence are observed.

Although an example has been shown in which excitation light of different wavelengths is irradiated sequentially, excitation light of the same wavelength may be irradiated repeatedly and sequentially, or the intensity may be changed and the light may be irradiated sequentially. Furthermore, in addition to irradiating multiple wavelengths simultaneously, multiple wavelengths may be irradiated sequentially so as to excite the light in order to raise the level. Furthermore, light may be irradiated using a combination of these.

In Variant 3 of the present invention, the sample is irradiated with light of multiple wavelengths or multiple intensities, or a combination of these, making it possible to obtain more information from a single sample. It is also possible to obtain the generation of multi-stage excited states and the corresponding reactions, which would not be possible with single light irradiation or light irradiation for a limited period of time.

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 a microchip as an example of the present invention, with the wavelength appropriately changed, to configure a particle separation device (cell sorter) equipped with a particle analysis device using the chip-type flow cell.

Figures 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.

The cell sorter 2B includes a flow cell 1B with a flat substrate as its housing and a flow path formed within the flat substrate; an excitation optical unit 11B as a light irradiation means that irradiates light onto particles in the sample liquid flowing through the flow path; a detector 730 as a detection means that detects scattered light, reaction light, or fluorescence of at least a specific wavelength emitted from the particles when irradiated with light; a microcomputer 790 as an analysis means that identifies particles and distinguishes target particles based on the signal intensity of these detection signals; and 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. The microcomputer 790 controls the operation of the electromagnetic valve based on the analysis results. The excitation optical unit 11B is configured to be either integrated with the flow cell 1B or can be attached without the need to adjust the position of the light irradiation area.

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. Examples of resins that can be used include polymetal acrylate (PMMA), cycloolefin copolymer (COC), and methylpentene polymer. Methylpentene polymer is particularly suitable as a flow cell material when using a laser in the UV wavelength range with a wavelength of approximately 350 nm to 410 nm as the irradiation light source. Wavelengths that can be selected include, for example, 405 nm, 488 nm, 561 nm, and 637 nm. In this case, the wavelengths of the light sources of the excitation optical unit 11B can be any combination of three wavelengths, for example, 405 nm, 488 nm, 561 nm, and 637 nm. The fluorescent 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 in a symmetrical structure, and the electromagnetic valve and cylinder pump are connected.

Figure 13(c) shows the connection relationship between the optical system and the control circuit. The cross-sectional view of the chip is taken along the line D-D in 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 along with the sheath liquid through the flow path 370. Light of a single or multiple wavelengths is irradiated from the excitation optical unit 11B onto the center of the main flow path, slightly upstream of the opposing separation flow paths 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 a 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, which serves as a data analysis unit. This completes the operation of the particle analyzer.

When used as a separation device, the data analysis unit further determines whether the analysis results meet preset separation conditions, and if so, outputs a trigger signal to the electromagnetic valve driver 760 after a fixed 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 fixed period of time. When the electromagnetic valve opens, the target cells flow into the separation reservoir 5-1 or 5-2. Sheath fluid and sample fluid that do not flow into the separation reservoir flow into the collection reservoir 210.

As shown in Figure 13(a), a constant air pressure is applied to the reservoir 300 upstream of the chip using 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. Due to the common air pressure, the sample fluid 20 flows 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 in a narrowed state due to 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 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 channel 370, located a few hundred microns upstream from this facing area, 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 50 μm long and 20 μm wide.

(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.

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

Figures 14(a) to (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. Note that Figure 14(a) is a plan view, Figure 14(b) is a front cross-sectional view when detecting transmitted light or scattered light, and Figure 14(c) is a plan cross-sectional view when detecting scattered light, fluorescence, or reaction luminescence.

The measurement system 3 includes a pump 4, a pump 5, a microchip 1C having a flow path 31 into which sample liquid A flows from pump 4, a flow path 32 into which sample liquid B flows from pump 5, and a flow path 33 where these flow paths converge; a recovery unit 6 for the converged liquid; an excitation optical unit 11C that irradiates the converged sample liquid in the microchip 1C with light of multiple wavelengths; a spectrometer 10C formed in the microchip 1C that disperses light incident from the sample using multiple filters; a detector 14 that detects the light characteristics corresponding to the irradiated light and detects the light dispersed by the spectrometer 10C as an analytical means for analyzing chemical changes; a signal processing unit 15; and a data analysis unit 16. The wavelengths used in the waveguide multiplexer and waveguide spectrometer satisfy the conditions for the multiplexer and spectrometer described above 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.

An example of operation is described below. Sample solution A, to which a protonated dye and an ionophore such as a 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 of target ions can be measured as an example of analyzing chemical changes that have occurred in a sample.

That is, as shown in FIG. 14(b), excitation optical unit 11C irradiates the dye in sample solution A with light of a wavelength selected to correspond 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 of multiple dyes (before, after, or both of the shifts) combined. Spectrometer 10C separates the light irradiated by excitation optical unit 11C into specific wavelengths corresponding to the light, and uses detector 14, signal processing unit 15, and data analysis unit 16 to detect the light intensity as an example of a characteristic quantity of the light passing through sample solution A and determine any changes in this intensity. Spectrometer 10C is positioned so that it can detect the light irradiated by excitation optical unit 11C and transmitted through sample solution A.

Alternatively, excitation light can be irradiated onto the dye in sample solution A, and as shown in FIG. 11(c), spectrometer 10C can be configured to receive scattered light, fluorescence, or reaction luminescence in a direction perpendicular to the direction of light irradiation, as in the first and second embodiments. Spectrometer 10C then separates the light into wavelengths corresponding to the light irradiated by excitation optical unit 11C, and detects the characteristic quantities of the light emitted from the dye in sample solution A using detector 14, signal processing unit 15, and data analysis unit 16.

(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.

Other Embodiments
The present invention is not limited to the above-described embodiment, and various modifications are possible without departing from the spirit of the present invention.

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

In addition, any known detector that can detect weak light can be used as the detector, such as an APD (Avalanche Photodiode). Examples of array detectors include an APD array and a multi-channel PMT (PhotoMultiplier Tube).

1: Microchip 1B: Flow cell 1C: Microchip 2: Flow cytometer 2B: Cell sorter 3: Measurement system 4, 5: Pump 6: Recovery 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 section 20: Sample liquid 24: Condenser lenses 31, 32, 33: Flow path 110: Light source 111: Lens 112: Substrate 113: MEMS mirror device

Claims (9)

a light irradiation means having a plurality of light sources and a 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 condensing member of the plurality of light sources has passed through the condensing member, and an area where the respective beams of the plurality of light sources after passing through the condensing member intersect is an optical irradiation area, wherein the light beams emitted from the emission spots located most centrally with respect to the condensing member of the plurality of light sources are light beams that spread in the light emission direction, and the beams immediately after passing through the condensing member are light beams whose width of the luminous flux of the light beams does not change in the light emission direction or light beams that converge in the light emission direction;
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 . The microchip described in claim 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. The microchip described in claim 2, wherein the mirror rotates to change its angle with respect to the light beam, and moves the light irradiation area in accordance with the flow of sample in the sample solution. The microchip described in claim 2 or 3, wherein the light irradiation means sequentially irradiates light beams of multiple wavelengths and/or multiple intensities. The microchip described in claim 3 or 4, wherein the mirror returns to its initial position from which it begins tracking when detection of transmitted light, reaction luminescence, fluorescence, or scattered light from the sample in the sample solution in the light irradiation area ceases. 2. The microchip according to claim 1, wherein the microchip and the light irradiation means have an embedding structure such that the light irradiation position of the light irradiation means coincides with the light irradiation region on the flow channel of the microchip, and the light irradiation means is installed on the microchip by the embedding structure. 2. The microchip according to claim 1, further comprising an attachment for alignment so that the light irradiation position of the light irradiation means coincides with the light irradiation region on the flow channel of the microchip, and the light irradiation means is installed on the microchip by the attachment. 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 path 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 that does not change in width of the luminous flux of the light beam in the direction of light emission or a light beam that converges in the direction of light emission . 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 whose luminous flux width does not change 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.