Microchips, sample collection kits, and microparticle collection devices

Abstract

To provide a microchip which is easy to handle.SOLUTION: A tabular microchip includes: a sample liquid inlet in which a sample liquid is introduced; a main flow passage in which the sample liquid introduced from the sample liquid inlet flows; and a separation flow path in which a target sample is separated from the sample liquid, the terminal ends of the sample liquid inlet and the separation flow path being formed in one side surface. There are also provided a sample separation kit having the microchip, and a small particle sample separation device with the microchip equipped thereto.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present technology relates to a microchip, a sample collection kit, and a microparticle sorting device. More specifically, the present technology relates to an easy-to-handle microchip, a sample collection kit including the microchip, and a microparticle sorting device equipped with the microchip. [Background technology]

[0002] Various devices have been developed to collect microparticles. For example, in a microparticle collection system used in a flow cytometer, a laminar flow consisting of a sample liquid containing microparticles and a sheath liquid is ejected from an orifice formed in a flow cell or a microchip. During ejection, a predetermined vibration is applied to the laminar flow to form droplets. The movement direction of the formed droplets is electrically controlled depending on whether or not they contain the target microparticles, and the target microparticles are collected.

[0003] A technique for separating target microparticles in a microchip without forming droplets as described above has also been developed. For example, Patent Document 1 below describes "a microchip having a sample liquid introduction channel through which a sample liquid containing microparticles flows, at least one pair of sheath liquid introduction channels that merge with the sample liquid introduction channel from both sides and introduce sheath liquid around the sample liquid, a confluence channel that communicates with the sample liquid introduction channel and the sheath liquid introduction channel and through which the liquids flowing through these channels merge and flow together, a negative pressure suction unit that communicates with the confluence channel and sucks in microparticles to be collected, and at least one pair of waste channels that are provided on both sides of the negative pressure suction unit and communicate with the confluence channel." (Claim 1). In this microchip, target microparticles are collected into the negative pressure suction unit by suction. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Japanese Patent Application Laid-Open No. 2012-127922 Summary of the Invention [Problem to be solved by the invention]

[0005] In the structure of conventional microchips, the sample liquid inlet into which the sample liquid containing microparticles is introduced and the end of the sorting flow path from which the microparticles to be recovered from the sample liquid are sorted are formed on different sides. As a result, when inserting the microchip into a device, it is necessary to install a collection container, bag, etc. on the side corresponding to each side.

[0006] Therefore, a main object of the present technology is to provide a microchip that is easy to handle. [Means for solving the problem]

[0007] The present inventors have found that the above problems can be solved by a microchip having a specific configuration.

[0008] That is, the present technology provides a plate-shaped microchip having a sample liquid inlet into which a sample liquid is introduced, a main flow path through which the sample liquid introduced from the sample liquid inlet flows, and a separation flow path through which a target sample is separated from the sample liquid, with the ends of the sample liquid inlet and the separation flow path formed on the same side. The microchip according to the present technology may further include a sheath fluid inlet into which sheath fluid is introduced, the sheath fluid inlet being formed on the same side surface. In this case, the microchip may further include a buffer fluid inlet into which buffer fluid is introduced, the buffer fluid inlet being formed on the same side surface. In this case, the microchip may further include a branch channel branching from the main channel and discarding samples other than the target sample, the terminal of the branch channel being formed on the same side surface. Furthermore, the microchip according to the present technology may have a flow path connecting member inserted into at least one selected from the group consisting of the sample liquid inlet, the end of the separation flow path, the sheath liquid inlet, the buffer liquid inlet, and the end of the branch flow path. In this case, a protective part may be provided to protect the inserted flow path connecting member. In this case, the sample liquid flow path through which the sample liquid flows may have a suddenly expanded part at its end on the sample liquid inlet side, the cross-sectional area of ​​which is larger than the cross-sectional area of ​​the inner diameter of the flow path connecting member. Furthermore, the microchip according to the present technology may further include an orifice section coaxial with the main channel and connected to the sorting channel, and the sidewall of the sorting channel connected to the orifice section may have at least one curvature. In this case, the cross-sectional area of ​​the sorting channel may increase continuously along the direction of liquid flow up to a predetermined position. In this case, the sidewall of the sorting channel connected to the orifice section may have two different curvatures. In this case, the depth of the sorting channel may be constant up to a second curvature section, and the width may increase continuously along the direction of liquid flow up to the second curvature section. In this case, the depth of the sorting channel may increase continuously along the direction of liquid flow from the second curvature section onwards. Additionally, the sorting channel and the orifice portion may be formed in stacked substrate layers, with a portion of the sorting channel and / or a portion of the orifice portion being formed in a layer on one side of the substrate layer. In the microchip according to the present technology, at least a part of one surface of the substrate layer on which the sorting channel is formed may be exposed to the outside. Furthermore, the main channel may have a first optical detection region, and both surfaces of the substrate layer on which the first optical detection region is formed may be exposed to the outside. Additionally, the sorting channel may have a second optical detection region, and both surfaces of the substrate layer on which the second optical detection region is formed may be exposed to the outside.

[0009] The present technology also provides a sample fractionation kit having a sample liquid storage section that stores sample liquid, a sample liquid inlet into which the sample liquid is introduced, a main flow path through which the sample liquid introduced from the sample liquid inlet flows, and a fractionation flow path through which a target sample is fractionated from the sample liquid, and a plate-shaped microchip in which the ends of the sample liquid inlet and the fractionation flow path are formed on the same side, and in which the sample liquid storage section and the microchip are connected.

[0010] Furthermore, the present technology also provides a microparticle sorting device equipped with a plate-shaped microchip, which has a sample liquid inlet into which a sample liquid is introduced, a main flow path through which the sample liquid introduced from the sample liquid inlet flows, and a sorting flow path through which a target sample liquid is sorted from the sample liquid, and in which the ends of the sample liquid inlet and the sorting flow path are formed on the same side. The microparticle sorting device according to the present technology may include a chip insertion unit for inserting the microchip, a light irradiation unit for irradiating light onto the microparticles flowing through the main flow path, a light detection unit for detecting scattered light and / or fluorescence emitted from the microparticles, and a control unit for controlling the direction of travel of the microparticles flowing through the main flow path based on data detected by the light detection unit. In addition, the microparticle sorting device according to the present technology may include a sample sorting kit including a sample liquid storage section in which the sample liquid is stored and a sample liquid storage section connected to the microchip, and may further include a sample liquid delivery mechanism that delivers the sample from the sample liquid storage section to the microchip. [Brief explanation of the drawings]

[0011] [Figure 1] 1 is a front view showing a first embodiment of a microchip according to the present technology. [Figure 2] FIG. 1 is a rear view showing a first embodiment of a microchip according to the present technology. [Figure 3] FIG. 1 is a left side view showing a first embodiment of a microchip according to the present technology. [Figure 4]FIG. 1 is a right side view showing a first embodiment of a microchip according to the present technology. [Figure 5] 1 is a plan view showing a first embodiment of a microchip according to the present technology. [Figure 6] FIG. 1 is a bottom view showing a first embodiment of a microchip according to the present technology. [Figure 7] 1 is a front perspective view showing a first embodiment of a microchip according to the present technology. [Figure 8] 1 is a rear perspective view showing a first embodiment of a microchip according to the present technology. [Figure 9] 2 is an end view taken along line AA in FIG. 1 showing a first embodiment of the microchip according to the present technology. FIG. [Figure 10] FIG. 10 is an enlarged view of the CC portion of FIG. 9. [Figure 11] FIG. 10 is an enlarged view of the portion between DD in FIG. 9. [Figure 12] FIG. 10 is an enlarged view of the E-EE portion of FIG. 9. [Figure 13] 2 is an end view taken along line BB in FIG. 1 showing a first embodiment of a microchip according to the present technology. FIG. [Figure 14] FIG. 14 is an enlarged view of FIG. [Figure 15] FIG. 2 is a schematic cross-sectional view showing an example of the vicinity of a particle sorting section. [Figure 16] FIG. 16 is an end view of line FF in FIG. 15. [Figure 17] FIG. 2 is a schematic vertical cross-sectional view showing an example of the vicinity of an orifice portion. [Figure 18] FIG. 18 is a schematic vertical cross-sectional view showing an example near the orifice portion that is different from that shown in FIG. 17. [Figure 19] FIG. 2 is a schematic cross-sectional view showing an example of the vicinity of a sudden expansion portion of a sample liquid flow path. [Figure 20] 1 is a front perspective view of a first layer of a microchip according to a first embodiment of the present technology, represented by an opaque body. [Figure 21] 1 is a perspective view of a first layer of a microchip according to a first embodiment of the present technology, shown as an opaque body, from the rear side. [Figure 22]1 is a front perspective view of a second layer of a first embodiment of a microchip according to the present technology, the second layer being represented by an opaque body. [Figure 23] 1 is a rear perspective view of a second layer of a first embodiment of a microchip according to the present technology, the second layer being represented by an opaque body. [Figure 24] FIG. 2 is a front perspective view of a third layer of a first embodiment of a microchip according to the present technology, represented by an opaque body. [Figure 25] FIG. 2 is a rear perspective view of a third layer of a first embodiment of a microchip according to the present technology, which is represented by an opaque body. [Figure 26] FIG. 2 is a front perspective view of a fourth layer of a first embodiment of a microchip according to the present technology, the fourth layer being represented by an opaque body. [Figure 27] FIG. 2 is a rear perspective view of a fourth layer of a first embodiment of the microchip according to the present technology, represented by an opaque body. [Figure 28] FIG. 10 is a front view showing a second embodiment of the microchip according to the present technology. [Figure 29] FIG. 10 is a rear view showing a second embodiment of the microchip according to the present technology. [Figure 30] FIG. 10 is a left side view showing a second embodiment of the microchip according to the present technology. [Figure 31] FIG. 10 is a right side view showing a second embodiment of the microchip according to the present technology. [Figure 32] FIG. 10 is a plan view showing a second embodiment of the microchip according to the present technology. [Figure 33] FIG. 10 is a bottom view showing a second embodiment of the microchip according to the present technology. [Figure 34] FIG. 10 is a front perspective view of a second embodiment of a microchip according to the present technology, shown in an opaque body. [Figure 35] FIG. 10 is a rear perspective view of a second embodiment of a microchip according to the present technology, shown as an opaque body. [Figure 36] FIG. 10 is a front perspective view showing a third embodiment of the microchip according to the present technology. [Figure 37] 1 is a front view showing a configuration example of a sample collection kit according to the present technology. FIG. [Figure 38]1 is a rear view showing an example of the configuration of a sample collection kit according to the present technology. FIG. [Figure 39] 1 is a left side view showing an example of the configuration of a sample collection kit according to the present technology. FIG. [Figure 40] 1 is a bottom view showing an example of the configuration of a sample collection kit according to the present technology. FIG. [Figure 41] FIG. 10 is a front view showing another configuration example of the sample collection kit according to the present technology. [Figure 42] 1 is a diagram showing a configuration example of a microparticle sorting device according to the present technology; [Figure 43] FIG. 10 is a flow chart showing an example of attaching a sample fractionation kit to a microparticle fractionation device. DETAILED DESCRIPTION OF THE INVENTION

[0012] A preferred embodiment for carrying out the present technology will be described below. The embodiments described below are representative embodiments of the present technology, and should not be construed as narrowing the scope of the present technology. The present technology will be described in the following order. 1. First embodiment (microchip) 2. Second embodiment (microchip) 3. Third embodiment (microchip) 4. Fourth embodiment (sample collection kit) 5. Fifth embodiment (microparticle sorting device)

[0013] 1. First embodiment (microchip)

[0014] 1 to 14 are diagrams showing a first embodiment of a microchip 100 according to the present technology. The configuration of the microchip 100 according to this embodiment will be described below. Note that this embodiment shows a preferred example, and the microchip 100 according to the present technology is not limited to this configuration.

[0015] Microchip 100 according to this embodiment may have a channel structure as shown in Figures 1 to 14. In microchip 100 according to this embodiment, sample liquid inlet 101 and end 1091 of sorting channel 109 are formed on the same side. This allows microchip 100 to be easily inserted into and removed from a device (such as microparticle sorting device 300, which will be described later). Furthermore, when channel connecting members are inserted into sample liquid inlet 101 and end 1091 of sorting channel 109, the piping can be arranged in one direction, making the chip easy to handle.

[0016] The microchip 100 is also provided with a sample liquid inlet 101 into which a sample liquid is introduced and a sheath liquid inlet 103 into which a sheath liquid is introduced. In this embodiment, the sheath liquid inlet 103 is formed on the same side as the sample liquid inlet 101 and the end 1091 of the sorting channel 109. This prevents the handling of the microchip from becoming complicated.

[0017] A sample liquid and a sheath liquid are respectively introduced from a sample liquid inlet 101 and a sheath liquid inlet 103 into a sample liquid flow path 102 and a sheath liquid flow path 104. The sample liquid contains microparticles.

[0018] In the present technology, the sample liquid is not particularly limited as long as it is a specimen containing a target sample that can be separated using the microchip 100 according to the present technology. For example, it may be a liquid containing patient-derived cells, such as whole blood, peripheral blood mononuclear cells contained in whole blood, or a cell suspension containing only lymphocytes.

[0019] The sheath liquid flowing through the sheath liquid flow path 104 joins with the sample liquid flowing from both sides of the sample liquid flow path 102 at a joining point 111, forming a laminar flow in which the sample liquid is surrounded by the sheath liquid. This laminar flow flows through the main flow path 105 toward the particle sorting section 107.

[0020] The main channel 105 is provided with a first optical detection region 106. In the first optical detection region 106, light is irradiated onto microparticles in the sample liquid. Whether or not the microparticles should be collected can be determined based on the fluorescence and / or scattered light generated by the light irradiation. In this embodiment, both sides of the substrate layer on which the first optical detection region 106 is formed are exposed to the outside. This enables detection by the light detection unit 303, which will be described later.

[0021] In this case, the first optical detection region 106 may have a tapered shape on a portion of the wall forming the region, as shown in FIG. 11. This can prevent optical vignetting. The taper angle can be, for example, 5° to 30°, preferably 10° to 25°, and particularly preferably 15° to 20°. This can prevent any adverse effect on the bonding of the substrate layers forming the microchip 100. It can also be possible to arrange the first optical detection region 106 and the vibration region 1092 (described later) in close proximity to each other.

[0022] In this technology, one light may be irradiated onto one position in the first optical detection region 106, or light may be irradiated onto each of multiple positions in the first optical detection region 106. For example, the microchip 100 may be configured so that light is irradiated onto each of two different positions in the first optical detection region 106. That is, there may be two positions in the first optical detection region 106 where light is irradiated. In this case, for example, it may be determined whether a microparticle should be collected based on light (e.g., fluorescence and / or scattered light) generated by irradiating a microparticle with light at one position. Furthermore, the velocity of the microparticle in the flow channel may be calculated based on the difference between the detection time of light generated by irradiating the microparticle with light at one position and the detection time of light generated by irradiating the microparticle with light at another position. For this calculation, the distance between the two irradiation positions may be determined in advance, and the velocity of the microparticle may be determined based on the difference between the two detection times and the distance. Furthermore, the arrival time of the microparticle at the particle sorting unit 107, which will be described below, can be accurately predicted based on the velocity. Accurate prediction of the arrival time allows for optimization of the timing of the formation of a flow entering the sorting channel 109. Furthermore, if the difference between the arrival time of a certain microparticle at the particle sorting section 107 and the arrival time of a microparticle before or after the certain microparticle at the particle sorting section 107 is equal to or less than a predetermined threshold, it can be determined not to sort the certain microparticle. If the distance between the certain microparticle and the microparticle before or after the certain microparticle is small, the possibility of the certain microparticle being collected together with the previous or subsequent microparticle increases when the certain microparticle is aspirated. By determining not to sort the certain microparticle when there is a high possibility of the microparticles being collected together, it is possible to prevent the previous or subsequent microparticle from being collected. This increases the purity of the target microparticles among the collected microparticles. Specific examples of a microchip in which light is irradiated onto two different positions in the first optical detection region 106 and an apparatus including the microchip are described, for example, in JP 2014-202573 A.

[0023] In particle sorting section 107 in microchip 100, the laminar flow that has flowed through main channel 105 separates into two branch channels 108. Note that although particle sorting section 107 in the embodiment shown in Figures 1 to 14 has two branch channels 108, the number of branch channels is not limited to two. That is, particle sorting section 107 may be provided with, for example, one or more (e.g., two, three, or four) branch channels.

[0024] In the present technology, the branch channel 108 may be formed so that it branches into a Y-shape on a plane and then faces the side where the sample liquid inlet 101 and the end 1091 of the sorting channel 109 are located, as shown in Figures 1 to 14, or may be configured to branch three-dimensionally. In this embodiment, the end 1081 of the branch channel 108 is formed on the same side as the end 1091 of the sample liquid inlet 101 and the sorting channel 109. This improves the handling of the chip, and prevents cumbersome operations, for example, when inserting the chip into an apparatus.

[0025] In the particle sorting section 107, only when microparticles to be collected (also referred to as "target sample") flow in, a flow that enters the sorting channel 109 is formed, and the microparticles are collected. The flow that enters the sorting channel 109 can be formed, for example, by generating a negative pressure in the sorting channel 109. To generate the negative pressure, as in this embodiment, a vibration region 1092 is provided, and an actuator or the like can be attached to the outside of the microchip 100 so as to deform the wall of the region.

[0026] In this embodiment, one side of the substrate layer on which the vibration region 1092 is formed may be exposed to the outside. By exposing only one side to the outside in this manner, the rigidity of the microchip 100 itself can be increased and unnecessary vibrations can be reduced. The vibration region 1092 may have a tapered shape in part of the wall forming the region, similar to the first optical detection region 106 described above. Deformation of the wall of the region changes the internal space of the vibration region 1092, and negative pressure can be generated. The actuator may be, for example, a piezoelectric actuator. When the microparticles are sucked into the sorting channel 109, the sample liquid constituting the laminar flow or the sample liquid and sheath liquid constituting the laminar flow may also flow into the sorting channel 109. In this way, the microparticles to be collected may be collected.

[0027] FIG. 15 is a schematic cross-sectional view (a plane parallel to the front) showing an example of the vicinity of the particle sorting section 107. As shown in FIG. 15, the main channel 105 and the sorting channel 109 are connected via an orifice section 130 that is coaxial with the main channel 105. Microparticles to be collected flow through the orifice section 130 into the sorting channel 109. In addition, the orifice section 130 may be provided with a buffer liquid flow channel 110 to prevent microparticles that should not be collected from entering the sorting channel 109 through the orifice section 130. A buffer liquid is introduced from the buffer liquid flow channel 110, and a flow is formed by a portion of the introduced buffer liquid from the orifice section 130 toward the main channel 105, thereby preventing microparticles that should not be collected from entering the sorting channel 109.

[0028] The buffer solution inlet 1101, into which the buffer solution is introduced, is formed on the same side as the sample solution inlet 101 and the end 1091 of the sorting channel 109. This improves the handling of the chip, and prevents cumbersome operations, for example, when inserting the chip into an apparatus. Note that the remainder of the introduced buffer solution can flow into the sorting channel 109.

[0029] FIG. 17 is a schematic longitudinal cross-sectional view showing an example of the vicinity of the orifice portion 120. Note that this cross-sectional view is a schematic cross-sectional view of a plane passing through the center line of the buffer solution flow channel 110 and the center line of the orifice portion 120. The orifice portion 120 includes a flow channel 120a on the first optical detection region 106 side (hereinafter also referred to as the "upstream orifice flow channel 120a"), a flow channel 120b on the sorting flow channel 109 side (hereinafter also referred to as the "downstream orifice flow channel 120b"), and a connection portion 120c between the orifice portion 120 and the buffer solution flow channel 110. In this embodiment, the buffer solution flow channel 110 is disposed so as to be approximately perpendicular to the axis of the orifice portion 120. In this case, sufficient space can be secured near the orifice portion 120, the flow channels are not adjacent to each other, so the thickness of the flow channel wall can be maintained, and the bonding area of ​​the chip bonding surface can be increased, which is advantageous in terms of mechanical strength. In FIG. 17, two buffer liquid flow paths 110 are provided facing each other at approximately the center of the orifice portion 120, but only one buffer liquid flow path may be provided.

[0030] In this technology, various liquids can be selected as the buffer solution depending on the application. For example, liquids suitable for the microparticles can be selected, such as liquid media used in microparticle-containing liquids, sheath liquids, and buffer solutions containing surfactants and pH-adjusted surfactants when the microparticles are proteins. In particular, when the microparticles are cells, cell culture solutions, cell preservation solutions, etc. can be used. Cell culture solutions are suitable for subsequent processes of the target sample, such as cell culture, cell activation, and gene transfer. Cell preservation solutions are suitable for storing and transporting recovered cells. Furthermore, when the target sample is undifferentiated cells such as iPS cells, a differentiation-inducing solution can be used, allowing for efficient subsequent processing. Furthermore, a solution with a blocking effect can also be used as the buffer solution. This can suppress nonspecific adsorption of the target sample to the collection container or bag. Examples of blocking agents include solutions containing proteins such as albumin, solutions containing amino acids such as glycine, and solutions containing nonionic surfactants such as Pluronic F68. Furthermore, solutions with cytolytic properties can also be used as the buffer solution. This makes it possible to extract intracellular substances directly after separating the target sample population. Examples of cell lysis solutions include solutions containing surfactants.

[0031] Similarly, various liquids can be selected as the sheath liquid in the present technology. In this specification, the liquid flowing through the buffer liquid flow path 110 is referred to as the "buffer liquid."

[0032] In the present technology, the shape and dimensions of the cross section of the upstream orifice flow path 120a may be the same as the shape and dimensions of the downstream orifice flow path 120b. For example, the cross section of the upstream orifice flow path 120a and the cross section of the downstream orifice flow path 120b may both be substantially circular and have the same dimensions. Alternatively, both of these two cross sections may be rectangular (e.g., square, oblong, etc.) and have the same dimensions.

[0033] Furthermore, in the present technology, the shape and / or dimensions of the cross section of the upstream orifice section flow path 120a may be different from the shape and / or dimensions of the downstream orifice section flow path 120b. An example in which the dimensions of these two flow paths are different is shown in FIG. 18. As shown in FIG. 18, the orifice section 130 includes a flow path 130a on the first optical detection region 106 side (hereinafter also referred to as the "upstream orifice section flow path 130a"), a flow path 130b on the sorting flow path 109 side (hereinafter also referred to as the "downstream orifice section flow path 130b"), and a connection portion 130c between the orifice section 130 and the buffer solution flow path 110. Both the cross sections of the upstream orifice section flow path 130a and the downstream orifice section flow path 130b have a substantially circular shape, but the diameter of the latter cross section can be larger than the diameter of the former cross section. By making the diameter of the latter cross section larger than that of the former, it is possible to more effectively prevent microparticles already sorted into the sorting channel 109 from being released through the orifice section 130 into the main channel 105 immediately after the above-mentioned microparticle sorting operation by negative pressure, as compared to when the diameters of both are the same. For example, when the cross sections of the upstream orifice section channel 130a and the downstream orifice section channel 130b are both rectangular, by making the area of ​​the latter cross section larger than the area of ​​the former cross section, it is possible to more effectively prevent microparticles already collected from being released through the orifice section 130 into the main channel 105, as described above.

[0034] In this embodiment, a part of the orifice portion 130 can be formed on one of the substrate layers as shown in Fig. 18. This can reduce the influence of misalignment when the microchip 100 is made by bonding together multiple substrate layers. In particular, a portion of the sorting channel 109 may also be formed on one of the substrate layers, as shown in Figure 12. In particular, the orifice portion 130 may be configured to be closer to the substrate layer. This reduces the effect of the edge of the sorting channel 109 on deteriorating signal characteristics.

[0035] Figure 16 is an end view taken along line FF in Figure 15. In this embodiment, as shown in Figure 15, the side wall of the sorting channel 109 on the side connecting to the orifice section 130 can have at least one curvature. If the side wall does not have the curvature, the distance between the orifice section 130 and the vibration region 1092 of the sorting channel 109 becomes short, which places constraints on the design of the device and causes problems such as interference between a component (e.g., an objective lens) constituting the light detection unit 303 and an actuator, making it impossible to position either one.

[0036] 15 and 16, the cross-sectional area of ​​the sorting channel 109 can be increased continuously along the direction of the liquid flow up to a predetermined position (see point K in FIGS. 15 and 16), thereby increasing the flow rate near the orifice 130 and improving the sorting accuracy.

[0037] Furthermore, the sidewall of the sorting channel 109 particularly has two different curvatures. In this case, the first curvature in the direction of liquid flow (see point G in FIGS. 15 and 16 ) may be smaller than the second curvature in the same direction of liquid flow (see point H in FIGS. 15 and 16 ). For example, the first curvature may be φ1 mm or less, preferably φ0.5 mm or less, and the second curvature may be φ0.1 mm or more, preferably φ0.3 mm or more. In this way, by having the sidewall of the sorting channel 109 have two different curvatures, pressure loss and advection near the orifice 130 can be reduced, improving sorting accuracy.

[0038] 15 and 16, the depth of the sorting channel 109 is constant up to the second curvature portion (see point H in FIGS. 15 and 16), and the width can continuously increase along the direction of the liquid flow up to the second curvature portion, thereby increasing the flow rate near the orifice portion 130 and improving the sorting accuracy.

[0039] Furthermore, after the second curvature portion (see point H in FIGS. 15 and 16), the depth of the sorting channel 109 continuously increases along the direction of the liquid flow until the predetermined position (point K in FIGS. 15 and 16). This reduces pressure loss and advection near the orifice 130, improving sorting accuracy.

[0040] Furthermore, after the predetermined position, the depth of the sorting channel 109 may continuously increase along the direction of liquid flow, and the width may be constant up to a second predetermined position (see point L in Figure 16), or the width may also continuously increase along the direction of liquid flow.

[0041] As explained above, in this embodiment, by devising the shape of the flow path immediately after the orifice section 130, it is possible to extend the distance between the orifice section 130 and the sorting flow path 109, particularly the vibration region 1092, without impairing the sorting characteristics, thereby reducing the constraints on the device side.

[0042] The laminar flow that has flowed into branch channel 108 can be discharged to the outside of microchip 100 at terminal end 1081 of branch channel 108. Furthermore, the microparticles that have been collected into sorting channel 109 can be discharged to the outside of the microchip at terminal end 1091 of the sorting channel. In this way, the target sample is sorted by microchip 100.

[0043] 1 to 14, the microchip 100 according to the present technology may have a second optical detection region 1093 in the sorting channel 109. Light is irradiated onto the second optical detection region 1093. Whether or not the microparticles to be collected have been collected can be determined based on the fluorescence and / or scattered light generated by the light irradiation. In this embodiment, both surfaces of the substrate layer on which the second optical detection region 1093 is formed are exposed to the outside. This enables detection by the light detection unit 303, which will be described later. The second optical detection region 1093 may have a tapered shape in part of the wall forming the region, similar to the first optical detection region 106 described above.

[0044] In the microchip 100 according to the present technology, a channel connecting member may be inserted into at least one selected from the group consisting of the sample liquid inlet 101, the end 1091 of the sorting channel 109, the sheath liquid inlet 103, the buffer liquid inlet 1101, and the end 1081 of the branch channel 108. In particular, as shown in FIGS. 1 to 14 , channel connecting members (e.g., tubes) T1 to T5 may be inserted into all of the sample liquid inlet 101, the end 1091 of the sorting channel 109, the sheath liquid inlet 103, the buffer liquid inlet 1101, and the end 1081 of the branch channel 108. This can prevent sample retention, compared to, for example, connecting to a channel outside the microchip via a conventional manifold. Furthermore, in this case, the microchip 100 according to the present technology may have a structure for inserting each channel connecting member at its side end, as shown in FIGS. 1 to 14 .

[0045] The material of the tube serving as the flow path connecting member may be appropriately selected by those skilled in the art from those used in the art. The tube may be, for example, a polyvinyl chloride (PVC) tube, a silicone tube, a polyetheretherketone (PEEK) tube, a polytetrafluoroethylene (PTFE) tube, or a thermoplastic elastomer tube, or multiple types of tubes may be connected together.

[0046] The method for fixing each channel connecting member is not particularly limited, and examples thereof include a mechanical fitting method and a chemical bonding method, but in particular, they can be fixed with an adhesive, which can reduce the manufacturing cost of the microchip 100.

[0047] In this case, particularly, sample liquid flow path 102 through which the sample liquid flows may have a sudden expansion section 1021 at the end on the sample liquid inlet 101 side, the cross-sectional area of ​​which is larger than the cross-sectional area of ​​the inner diameter of the flow path connecting member T1, as shown in Figure 19. This is because if the cross-sectional area of ​​sample liquid flow path 102 is smaller than the cross-sectional area of ​​the inner diameter of the flow path connecting member T1, sample retention will occur at the end of flow path connecting member T1 on the sample liquid inlet 101 side. In particular, the shape of sudden expansion section 1021 can be such that the width of sample liquid inlet 101 is suddenly widened and then the width of the flow path is gradually narrowed, as shown in Figure 19. This prevents sample retention.

[0048] Furthermore, in the present technology, as shown in a fourth embodiment described later, a further flow path connecting member (for example, a tube) may be provided at the end of each flow path connecting member on the side not fixed to the microchip 100. In this case, in particular, a further flow path connecting member is inserted into the flow path connecting members T1 to T5, and the periphery thereof is fixed with, for example, an adhesive, etc., resulting in a structure in which the flow paths are aligned in the same straight line.

[0049] In the present technology, the term "micro" means that at least a portion of the channels included in the microchip has dimensions on the order of μm, particularly cross-sectional dimensions on the order of μm. That is, in the present technology, the term "microchip" refers to a chip including channels on the order of μm, particularly a chip including channels with cross-sectional dimensions on the order of μm. For example, a chip including a particle sorting section configured with channels with cross-sectional dimensions on the order of μm may be referred to as a microchip according to the present technology. For example, the cross section of the confluence channel 105 in the particle sorting section 107 may be rectangular, and the width d of the confluence channel 105 within the particle sorting section 107 may be, for example, 100 μm to 500 μm, particularly 100 μm to 300 μm. The width of the branch channel 108 branching from the confluence channel 105 may be smaller than the width of the confluence channel 105. The cross section of the orifice 130 is, for example, circular, and the diameter of the orifice 130 at the connection between the orifice 130 and the junction channel 105 can be, for example, 10 μm to 60 μm, particularly 20 μm to 50 μm. These dimensions of the channel may be changed as appropriate depending on the size of the microparticles, particularly the size of the target sample.

[0050] The microchip 100 according to the present technology can be manufactured by a method known in the art. For example, the microchip 100 can be manufactured by bonding two or more substrates on which predetermined channels are formed. The channels may be formed, for example, in all of the two or more substrates (particularly, two substrates), or may be formed in only some of the two or more substrates (particularly, one of the two substrates). Furthermore, the microchip 100 may be formed from three or more substrates (particularly, four substrates) by bonding additional substrates from above, below, or both directions relative to the plane of the substrate on which each channel is formed.

[0051] Materials known in the art can be used to form the microchip 100. Examples include, but are not limited to, polycarbonate, cycloolefin polymer, polypropylene, PDMS (polydimethylsiloxane), polymethyl methacrylate (PMMA), polyethylene, polystyrene, glass, and silicon. Polymer materials such as polycarbonate, cycloolefin polymer, and polypropylene are particularly preferred because they are easy to process and can be manufactured inexpensively using a molding device.

[0052] 1 to 14, microchip 100 is preferably transparent. For example, at least the portion of microchip 100 through which light (laser light and scattered light) passes is transparent, and for example, particle sorting section 107 may be transparent, but microchip 100 as a whole may also be transparent.

[0053] In the present technology, the "sample" contained in the sample liquid is particularly microparticles, and the microparticles may be particles having a size that allows them to flow through the flow path in the microchip 100. In the present technology, the microparticles may be appropriately selected by those skilled in the art. In the present technology, examples of the microparticles include biological microparticles such as cells, cell clumps, microorganisms, and liposomes, as well as synthetic microparticles such as gel particles, beads, latex particles, polymer particles, and industrial particles. Biological microparticles (also referred to as "bioparticles") may include chromosomes, liposomes, mitochondria, organelles, and the like that constitute various cells. Cells may include animal cells (e.g., blood cells) and plant cells. The cells may be, in particular, blood cells or tissue cells. The blood cells may be, for example, suspension cells such as T cells and B cells. The tissue cells may be, for example, adherent cultured cells or adherent cells dissociated from tissue. Cell aggregates may include, for example, spheroids and organoids. Microorganisms may include bacteria such as Escherichia coli, viruses such as tobacco mosaic virus, and fungi such as yeast. Furthermore, the biological microparticles may also include biological macromolecules such as nucleic acids, proteins, and complexes thereof. These biological macromolecules may be, for example, extracted from cells or contained in a blood sample or other liquid sample. The synthetic microparticles may be, for example, microparticles made of organic or inorganic polymeric materials or metals. Organic polymeric materials may include polystyrene, styrene-divinylbenzene, polymethyl methacrylate, etc. Inorganic polymeric materials may include glass, silica, magnetic materials, etc. Metals may include gold colloids, aluminum, etc. The synthetic microparticles may be, for example, gel particles, beads, etc., and in particular, gel particles or beads to which one or a combination of two or more selected from oligonucleotides, peptides, proteins, and enzymes is bound.

[0054] The shape of the microparticles may be spherical or nearly spherical, or may be non-spherical. The size and mass of the microparticles can be appropriately selected by those skilled in the art depending on the size of the flow path of the microchip 100. On the other hand, the size of the flow path of the microchip 100 can also be appropriately selected depending on the size and mass of the microparticles. In this technology, chemical or biological labels, such as fluorescent dyes and fluorescent proteins, can be attached to the microparticles as needed. The labels can make the detection of the microparticles easier. The labels to be attached can be appropriately selected by those skilled in the art. Molecules that specifically react with the microparticles (e.g., antibodies, aptamers, DNA, RNA, etc.) can be bound to the labels. In the present technology, the microparticles are preferably biological particles, and in particular may be cells.

[0055] The microchip 100 according to the present technology described above, in any of the present embodiment and the second and third embodiments described below, may be distributed as a single microchip 100 on the premise that it is connected in a sterile manner, or may be distributed as a component constituting a cartridge, unit, device, kit, instrument, or the like for a closed-type cell sorter, with the sample liquid storage section 201 or the like connected to a part of it.

[0056] 2. Second embodiment (microchip)

[0057] 28 to 35 are diagrams showing a second embodiment of the microchip 100 according to the present technology. The configuration of the microchip 100 according to this embodiment will be described below. Note that this embodiment shows a preferred example, and the microchip 100 according to the present technology is not limited to this configuration. In addition, in this embodiment, the configuration other than the protection unit 150 is the same as that of the first embodiment described above.

[0058] In this embodiment, the microchip 100 has flow path connecting members T1 to T5 inserted into the sample liquid inlet 101, the end 1091 of the sorting flow path 109, the sheath liquid inlet 103, the buffer liquid inlet 1101, and the end 1081 of the branch flow path 108, respectively, and has a protective part 150 that protects these flow path connecting members. The presence of the protective part 150 makes it possible to reduce bending stress on the flow path connecting members T1 to T5, for example, when the microchip 100 is inserted into or removed from a microparticle sorting device 300, which will be described later. In addition, it is also possible to reduce bending stress on flow path connecting members that may be further connected to the flow path connecting members T1 to T5.

[0059] 28 to 35, the protective part 150 may have a recess in a part of its side surface. The recess may be provided particularly on both side surfaces. This allows the recess to function as a handle for the microchip 100.

[0060] Furthermore, the microchip 100 according to the present technology may have a mechanism for preventing reverse insertion when inserted into an apparatus (such as the microparticle sorting apparatus 300 described below). In this case, for example, the protective part 150 may be used as the mechanism. The protective part 150 has a convex part 151, particularly on the front side. The convex part 151 may be continuous in the longitudinal direction of the microchip 100. The convex part 151 allows the chip to be inserted into the chip insertion part of the apparatus when inserted in the correct orientation, and when inserted in the wrong orientation, the convex part 151 catches on the apparatus, preventing insertion into the chip insertion part.

[0061] 3. Third embodiment (microchip)

[0062] FIG. 36 is a diagram showing a third embodiment of the microchip 100 according to the present technology. The configuration of the microchip 100 according to this embodiment will be described below. Note that this embodiment shows a preferred example, and the microchip 100 according to the present technology is not limited to this configuration. Furthermore, this embodiment is similar to the first embodiment described above, except that it does not have the buffer solution inlet 1101 and the buffer solution flow path 110, as compared to the first embodiment described above.

[0063] In this embodiment, the target sample is collected, for example, as follows.

[0064] In the microchip 100 according to this embodiment, a sample liquid inlet 101 and a sheath liquid inlet 103 are provided on the same side. From these inlets, the sample liquid and the sheath liquid are introduced into a sample liquid flow path 102 and a sheath liquid flow path 104, respectively. The sample liquid contains microparticles.

[0065] The sheath liquid flowing through the sheath liquid flow channel 104 merges with the sample liquid flowing from both sides of the sample liquid flow channel 102, forming a laminar flow in which the sample liquid is surrounded by the sheath liquid. This laminar flow flows through the main channel 105 toward the particle sorting section 107.

[0066] In particle sorting section 107, the laminar flow that has flowed through main channel 105 flows into branch channel 108. Furthermore, only when microparticles to be collected flow into particle sorting section 107, a flow into sorting channel 109 is formed, and the microparticles are collected. When the microparticles are sucked into sorting channel 109, the sample liquid that constitutes the laminar flow, or the sample liquid and sheath liquid that constitute the laminar flow, may also flow into sorting channel 109. In this way, the microparticles are sorted in particle sorting section 107.

[0067] In this embodiment, when microparticles are collected, a flow is formed that proceeds from the main channel 105 through the orifice portion 130 to the particle sorting channel 109, but compared to the first embodiment described above, the accuracy of sorting the microparticles to be collected is reduced because the present embodiment does not have the buffer liquid inlet 1101 and the buffer liquid channel 110. In the first embodiment described above, the presence of the buffer liquid inlet 1101 and the buffer liquid channel 110 makes it possible to prevent microparticles that should not be collected from passing through the orifice portion 130 and entering the sorting channel 109.

[0068] 4. Fourth embodiment (sample collection kit)

[0069] 37 to 40 are diagrams showing configuration examples of a sample collection kit 200 according to the present technology. The configuration of the sample collection kit 200 according to this embodiment will be described below. Note that this embodiment shows a preferred example, and the sample collection kit 200 according to the present technology is not limited to this configuration. In addition, the microchip 100 is the same as that described above, so a description thereof will be omitted here.

[0070] A sample liquid containing microparticles to be collected is stored in the sample liquid storage section 201. In the sample fractionation kit 200 according to the present technology, the sample liquid storage section 201 and the microchip 100 according to the present technology are connected to each other. In particular, they are connected in a sealed manner.

[0071] The sample liquid storage section 201 can be formed, for example, from a cylindrical body with one open end and a lid that fits into the cylinder and closes the opening. The lid is formed with a plurality of opening valves for storing the sample liquid inside the cylinder, and each opening valve is configured as a check valve. Therefore, when the sample liquid is stored in the sample liquid storage section 201 through the opening valves, the sample liquid does not leak out of the sample liquid storage section 201. Furthermore, the configuration of the opening valves seals the sample liquid from the external atmosphere.

[0072] In the present technology, a substance that suppresses aggregation of microparticles in the sample liquid may be provided in sample liquid storage section 201. By using a substance that suppresses aggregation of microparticles in the sample liquid, aggregation of particles in the sample liquid can be suppressed, and any aggregates that still occur can be removed by filter section 202, which will be described later, so that impurities in the sample liquid can be removed more reliably.

[0073] The sample liquid storage section 201, the pre-sample storage section 2011, the target sample reservoir 203, the waste section 204, the sheath liquid storage section 205, and the buffer liquid storage section 206, which will be described later, may be soft containers such as plastic bags. The plastic bags may be made of polyethylene, polypropylene, polyvinyl chloride, or ethylene-vinyl acetate copolymer, for example.

[0074] Furthermore, in the present technology, the sample liquid storage section 201 and the target sample storage section 203 described later may be not only the bag-shaped soft container described above, but also a tube-shaped hard container such as a test tube, as shown in another configuration example in Figure 41.

[0075] In addition, in the particle sorting kit 200 according to the present technology, a pre-sample storage section 2011 may be provided upstream of the sample liquid storage section 201, and the pre-sample storage section 2011 may contain a substance that suppresses aggregation of microparticles in the sample liquid.

[0076] The filter section 202 includes at least a filter and a tapered section, and may include, as necessary, a fitting section that fits in the outer diameter with a flow path connecting member for connecting to the sample liquid storage section 201 and / or the microchip 100. This makes it possible to prevent microparticles in the sample liquid that has passed through the filter from settling on the inner wall surface of the filter section 202, thereby reducing the amount of loss of microparticles.

[0077] The filter section 202 can be placed at any position appropriate to those skilled in the art, but for example, as shown in Figure 37, by placing it upstream of the sample liquid storage section 201, it is possible to prevent foreign matter from entering the sample liquid storage section 201 at an early stage.

[0078] 37, a filter section 202 may be disposed between sample liquid storage section 201 and microchip 100. In particular, filter section 202 may be disposed immediately before microchip 100. This makes it possible to reliably prevent foreign matter from entering microchip 100, and improves the accuracy of separating the target sample within microchip 100.

[0079] Microparticles to be collected are stored in the target sample storage section 203. The target sample storage section 203 is formed, for example, in a bag shape, and includes an opening valve connected to the end 1091 of the sorting channel 109 of the microchip 100. The opening valve employs a so-called check valve configuration, and when the microparticles to be collected are stored in the target sample storage section 203 via the opening valve, the microparticles do not escape to the outside of the target sample storage section 203. Furthermore, the configuration of the opening valve prevents the microparticles from coming into contact with the external atmosphere. The above-described configuration of the target sample reservoir 203 is merely an example, and any known configuration can be adopted as long as the target sample does not come into contact with the external atmosphere.

[0080] In the sample fractionation kit 200 according to the present technology, when fractionating only the target sample from the sample liquid using the microchip 100 described above, it is necessary to eliminate microparticles that should not be collected (hereinafter also referred to as "non-target samples"). Furthermore, because the target sample is fractionated by forming a sheath flow in the microchip 100, it is necessary to eliminate the sample liquid containing the non-target samples, i.e., so-called waste liquid. For this reason, the sample fractionation kit 200 may be provided with a disposal unit 204. Non-target samples other than the target sample can be discarded in the disposal unit 204.

[0081] The waste section 204 may include, for example, a channel connecting member for allowing waste liquid to flow in, and the member may be configured to communicate with the end 1081 of the branch channel 108 of the microchip 100. This allows the target sample to be separated and non-target samples to be discarded within the sealed space including the waste section 204.

[0082] Furthermore, a sheath flow is formed in the microchip 100 to separate a target sample from the sample liquid. For this reason, the sample separation kit 200 may include a sheath liquid storage unit 205. The sheath liquid storage unit 205 may store sheath liquid.

[0083] The sheath fluid storage section 205 may include, for example, a channel connecting member through which the sheath fluid flows, and the member may be configured to communicate with the sheath fluid inlet 103 of the microchip 100. This allows the sheath fluid to flow into the sheath fluid channel 104 of the microchip 100, forming a sheath flow.

[0084] There are no particular limitations on the configuration of the sheath fluid storage unit 205, and any known configuration may be employed. In addition, there are no particular limitations on the configuration for discharging the sheath fluid from the sheath fluid storage unit 205, and for example, a drive source such as an actuator may be used. A buffer solution is stored in the buffer solution storage section 206. The buffer solution is the same as that described above, and therefore a description thereof will be omitted here.

[0085] The buffer solution storage section 206 may include, for example, a channel connecting member into which the buffer solution flows, and the member may be configured to communicate with the buffer solution inlet 1101 of the microchip 100. This allows the buffer solution to flow into the channel of the microchip 100, and the target sample is collected.

[0086] There are no particular limitations on the configuration of the buffer solution storage unit 206, and any known configuration can be used. In addition, there are no particular limitations on the configuration for discharging the buffer solution from the buffer solution storage unit 206, and for example, a drive source such as an actuator can be used.

[0087] In the present technology, the sheath fluid storage unit 205 and the buffer fluid storage unit 206 may be configured as a common storage unit, as shown in another configuration example in FIG. 41 . Specifically, for example, an embodiment may be adopted in which the sheath fluid and the buffer fluid are supplied from a single reagent bag. In this case, it is not necessary to provide a sample fluid supply mechanism 305 (described later) for each of the sheath fluid storage unit 205 and the buffer fluid storage unit 206; as shown in FIG. 41 , it is sufficient to provide one sample fluid supply mechanism 305 for each of the single reagent bags. In this case, the flow paths may be branched by the sample fluid supply mechanism 305, and the amounts of sheath fluid and buffer fluid may be adjusted by the flow path resistance within the microchip 100 (for example, the diameter of the microchip 100, the diameter of each flow path, etc.).

[0088] This embodiment may include a damper 207 that reduces pulsation and a closed-type pressure sensor 208 that detects the liquid delivery pressure. For example, when a part or all of the liquid in the sample collection kit 200 is delivered by a pump, fluctuations in flow rate (e.g., pulsation) caused by the pump can affect the flow rate in the microchip 100, particularly the flow rate in the sorting channel 109, and the sorting of microparticles in the particle sorting section 107. Therefore, the damper 207 can be provided to reduce such effects and keep the pressure due to the liquid delivery as constant as possible. Furthermore, in this case, as shown in this embodiment, a pressure sensor 208 that measures the pressure can be provided for each damper 207. This allows for stable liquid delivery to each section. The damper 207 and the pressure sensor 208 can be disposed downstream of the sheath liquid storage section 205 and / or the buffer liquid storage section 206 and between the microchip 100 and the damper 207. In addition, in the present technology, the damper 207 and the pressure sensor 208 do not necessarily have to be provided together, and either one of them may be provided.

[0089] In this embodiment, the sample collection kit 200 may include the damper 207, the pressure sensor 208, and a portion of the flow path connecting member in a plate-like structure, as shown in Figures 37 to 41. The plate-like structure may be appropriately selected by a person skilled in the art from structures employed in the art. Materials known in the art may be used to form the plate-like structure. Examples include, but are not limited to, polycarbonate, cycloolefin polymer, polypropylene, PDMS (polydimethylsiloxane), polymethyl methacrylate (PMMA), polyethylene, polystyrene, glass, and silicon.

[0090] The components of the sample collection kit 200 according to the present technology, including the microchip 100, may all be connected from the beginning as shown in Figures 37 to 40, or may be configured so that some of them are connected in a sterile manner later as shown in Figure 41. The method of connecting them in a sterile manner later can be performed by using a sterile welder, a sterile connector, or the like.

[0091] By using the sample fractionation kit 200 according to the present technology, the fractionation and storage of the target sample can be performed in an enclosed space, thereby improving the accuracy of fractionation of the target sample. Furthermore, contamination of the sample fractionation kit itself by mist containing the target sample and / or contamination of the fractionated target sample with other substances can be prevented. Therefore, the sample fractionation kit 200 according to the present technology can also be applied to clinical applications such as immune cell therapy, which require high purity of the target sample.

[0092] Furthermore, the sample collection kit 200 itself can be made disposable, which can avoid the risk of contamination between samples and improve usability.

[0093] Furthermore, the sample collection kit 200 may have a structure that engages with an attachment portion on the device when attached to a microparticle collection device 300 (described later) or the like. For example, a hook may be provided on the device and a hole that engages with the hook may be provided in a corner of the kit, but this may be appropriately selected by a person skilled in the art from structures employed in this technical field.

[0094] A plurality of each of the above-described components of the sample collection kit 200 may be provided. For example, although not shown, it is possible to provide a microchip 100 downstream of the target sample storage unit 203, and to further collect the target sample collected from the sample liquid.

[0095] 5. Fifth embodiment (microparticle sorting device)

[0096] FIG. 42 is a diagram showing an example of the configuration of a microparticle sorting device 300. As shown in FIG. The configuration of the microparticle sorting device 300 according to this embodiment will be described below. Note that this embodiment shows a preferred example, and the microparticle sorting device 300 according to the present technology is not limited to this configuration. Furthermore, the microchip 100 and the sample sorting kit 200 are the same as those described above, and therefore a description thereof will be omitted here.

[0097] The microparticle sorting device 300 according to the present technology is equipped with the above-described microchip 100. As shown in Fig. 42, the device may have a chip insertion part 301 into which the microchip 100 is inserted. The chip insertion part 301 may be appropriately selected by a person skilled in the art from structures employed in the technical field, as long as it has a structure into which a chip can be inserted.

[0098] The chip insertion unit 301 may have a presence sensor that reacts only when the microchip 100 is inserted in the correct orientation. When the presence sensor reacts, the chip insertion unit 301 may automatically clamp the microchip 100 in the insertion direction. This prevents the chip from being inserted backwards.

[0099] Furthermore, when microchip 100 has protective part 150, a part of chip insertion part 301 can engage with a part of protective part 150 to temporarily position the chip. For example, a recess can be provided at the end of protective part 150, and this recess can engage with a ball plunger on the chip insertion part 301 side to prevent the chip from coming off. This gives the user a clicking sensation, notifying them that the chip has been inserted completely, and also applies tension to the channel connecting member, preventing the chip from coming off chip insertion part 301.

[0100] In this embodiment, the microparticle sorting device 300 may have a light irradiation unit 302 that irradiates light onto microparticles flowing through the first optical detection region 106 in the microchip 100, and a light detection unit 303 that detects scattered light and / or fluorescence generated by the light irradiation. In addition, the light detection unit 304 may irradiate the second optical detection region 1093 in the microchip 100 with light.

[0101] The microparticle sorting device 300 may also include a control unit 304. The control unit 304 controls the direction of travel of the microparticles flowing through the main channel 105 based on data detected by the light detection unit 303 (e.g., information related to light, etc.). The light irradiating unit 302, the light detecting unit 303, and the control unit 304 will be described below.

[0102] The light irradiation unit 302 irradiates light (e.g., excitation light) onto microparticles flowing through the first optical detection region 106 in the microchip 100. The light irradiation unit 302 may include a light source that emits light and an objective lens that focuses the excitation light onto the microparticles flowing through the detection region. The light source may be appropriately selected by a person skilled in the art depending on the purpose of sorting, and may be, for example, a laser diode, an SHG laser, a solid-state laser, a gas laser, or a high-intensity LED, or a combination of two or more of these. In addition to the light source and the objective lens, the light irradiation unit 302 may include other optical elements as needed. For example, the light irradiation unit 302 may irradiate light onto one position in the first optical detection region 106, or may irradiate light onto each of multiple positions. For example, the light irradiation unit 302 may irradiate light onto each of two different positions in the first optical detection region 106.

[0103] The light detection unit 303 detects scattered light and / or fluorescence generated from the microparticles upon irradiation by the light irradiation unit 302. The light detection unit 303 may include a condenser lens that condenses the fluorescence and / or scattered light generated from the microparticles and a detector. The detector may be, but is not limited to, a PMT, a photodiode, a CCD, a CMOS, or the like. The light detection unit 303 may include other optical elements as needed in addition to the condenser lens and the detector. The light detection unit 303 may further include, for example, a spectroscopic unit. Examples of optical components that constitute the spectroscopic unit include a grating, a prism, and an optical filter. The spectroscopic unit can, for example, separate and detect light of a wavelength to be detected from light of other wavelengths.

[0104] The fluorescence detected by the light detection unit 303 may be, but is not limited to, fluorescence generated from the microparticle itself and fluorescence generated from a substance labeled on the microparticle, such as a fluorescent substance. The scattered light detected by the light detection unit 303 may be forward scattered light, side scattered light, Rayleigh scattering, Mie scattering, or a combination thereof.

[0105] The control unit 304 controls the direction of travel of the microparticles flowing through the main channel 105 based on data (e.g., information about light) detected by the light detection unit 303. For example, the control unit 304 controls the sorting of the microparticles based on the data. For example, the control unit 304 may determine to sort the microparticles if the light detected by the light detection unit 303 satisfies a predetermined criterion. Information about the light (fluorescence and / or scattered light) detected by the light detection unit 303 may be generated from the light. The information may be generated, for example, by converting the light into an electrical signal. To generate the information, the microparticle sorting device 300 of the present technology may include an information generation unit that generates information about the light from the light detected by the light detection unit 303. The information generation unit may be included in the control unit 304, or may not be included in the control unit 304 and may be provided in the microparticle sorting device 300 as a component separate from the control unit 304. Based on the information about the light, the control unit 304 can determine whether the light detected by the light detection unit 303 satisfies a predetermined criterion. Based on the result of the determination, the control unit 304 can control the sorting of the microparticles.

[0106] If the microparticles are to be collected based on the result of this determination, the controller 304 may change the flow in the channel so that the microparticles pass through the orifice and proceed into the sorting channel 109. This change in flow may be performed, for example, by reducing the pressure in the sorting channel 109. After the microparticles have been collected, the controller 304 may change the flow in the channel again. This change in flow may be performed by increasing the pressure in the particle sorting channel. That is, the controller 304 may control the pressure in the particle sorting channel based on information about the light detected by the light detection unit 303.

[0107] The control unit 304 may have the same function as the drive unit described in, for example, Japanese Patent Application Laid-Open No. 2014-036604. That is, the control unit 304 can control an actuator configured to generate negative pressure in the sorting channel 109. When it is determined based on the information about the light that microparticles should be collected, the control unit 304 drives the actuator to generate negative pressure in the sorting channel 109. As a result, the microparticles that should be collected are collected in the sorting channel 109. When it is determined based on the information about the light that the microparticles should not be collected, the control unit 304 does not drive the actuator. As a result, the microparticles that should not be collected flow into the branch channel 108.

[0108] The actuator may be, for example, a piezoelectric element such as a piezo element. When it is determined that microparticles should be collected, the controller 304 applies a voltage to the piezo element that causes piezo contraction, thereby increasing the volume in the sorting channel 109. This increase in volume generates a negative pressure in the sorting channel 109. As a result, a flow is formed from the main channel 105 to the sorting channel 109, and the microparticles are collected in the sorting channel 109. When it is determined that microparticles should not be collected, the controller 304 does not apply the voltage. As a result, no flow is formed in the sorting channel 109, and the microparticles flow into the branch channel 108.

[0109] In this embodiment, the microparticle sorting device 300 may include the above-described sample sorting kit 200. In this case, the microparticle sorting device 300 may include a sample liquid delivery mechanism 305 that delivers a sample from the sample liquid storage section 201 to the microchip 100, as shown in Fig. 42 . The sample liquid delivery mechanism 305 may particularly be a pump. In particular, the sample liquid delivery mechanism 305 may be disposed downstream of the sample storage section 201 and between the sample storage section 201 and the microchip 100. The pump may be, for example, but not limited to, a peristaltic pump (tube pump), a roller pump, a syringe pump using an air pressure source as a compressor, or a centrifugal pump. The pump may be, in particular, a peristaltic pump or a roller pump for more precise control of the flow rate.

[0110] 37, a plurality of sample liquid delivery mechanisms 305 may be provided. Furthermore, the sample liquid delivery mechanism 305 may be further disposed downstream of the microchip 100 between the waste section 204, downstream of the sheath liquid storage section 205 between the microchip 100, or downstream of the buffer liquid storage section 206 between the microchip 100.

[0111] Furthermore, in this embodiment, the microparticle sorting device 300 may have a plurality of attachment parts to which each part of the sample sorting kit 200 can be attached. The structure of the attachment parts may be appropriately selected by a person skilled in the art from structures employed in this technical field.

[0112] Additionally, as described above, when the sample liquid storage unit 201 is a tubular hard container as shown in FIG. 41 , the present technology may fix the tubular hard container to, for example, a plate with holes, and mix the contents while vibrating them with an XYStage, and cool the hard container. In this case, cooling may be performed not only on the sample liquid storage unit 201 but also on the target sample reservoir 203. Examples of cooling methods include placing the sample liquid storage unit 201 and the target sample reservoir 203 in a refrigerator, or contacting them with a cooling element such as a Peltier element. The cooling mechanisms of the sample liquid storage unit 201 and the target sample reservoir 203 may be controlled individually or by the same control.

[0113] FIG. 43 is a flow diagram showing an example of attaching the sample collection kit 200 to the microparticle collection device 300 of this embodiment. The following describes the flow of attaching the sample collection kit 200 to the microparticle sorting device 300 according to this embodiment. Note that this flow shows a preferred example, and the attachment of the sample collection kit 200 to the microparticle sorting device 300 is not limited to this flow.

[0114] First, a part of the sample collection kit 200 (e.g., a part of the plate-like structure) is attached to an attachment part (e.g., a hook) on the device side (S11). Next, the disposal part 204 is placed in a tray on the device side (S12). Next, the target sample storage part 203 is attached to an attachment part on the device side (S13). Next, the sample liquid storage part 201 is attached to an attachment part on the device side (S14). Next, the microchip 100 in the sample collection kit 200 is inserted into the chip insertion part 301 (S15). Next, a part of the sample collection kit 200 (e.g., a part of the flow path connecting member) is attached to the sample liquid delivery mechanism 305 (S16). Next, the pressure sensor 208 is attached to an attachment part on the device side (S17). Next, the buffer liquid storage part 206 is attached to an attachment part on the device side (S18). Next, the sheath liquid storage part 205 is attached to an attachment part on the device side (S19). Finally, the pre-sample storage section 2011 is attached to the attachment section on the device side (S20).

[0115] The present technology can employ the following configuration. [1] a sample liquid inlet into which the sample liquid is introduced; a main flow path through which the sample liquid introduced from the sample liquid inlet flows; a separation flow path through which a target sample is separated from the sample liquid; and A plate-like microchip in which the sample liquid inlet and the end of the sorting channel are formed on the same side. [2] a sheath fluid inlet into which sheath fluid is introduced; The microchip according to [1], wherein the sheath liquid inlets are formed on the same side surface. [3] Further, the liquid supply system has a buffer liquid inlet into which a buffer liquid is introduced; The microchip according to [2], wherein the buffer solution inlets are formed on the same side surface. [4] a branch channel branching from the main channel and through which samples other than the target sample are discarded; The microchip according to [3], wherein the ends of the branch channels are formed on the same side surface. [5] The microchip according to [4], wherein a channel connecting member is inserted into at least one selected from the group consisting of the sample liquid inlet, the end of the separation channel, the sheath liquid inlet, the buffer liquid inlet, and the end of the branch channel. [6] The microchip according to [5], which has a protective part that protects the inserted channel connecting member. [7] The microchip according to [5] or [6], wherein the sample liquid flow path through which the sample liquid flows has a suddenly expanded section at the end on the sample liquid inlet side that has a cross-sectional area larger than the cross-sectional area of ​​the inner diameter of the flow path connecting member. [8] an orifice portion that is coaxial with the main channel and is connected to the sorting channel; The microchip according to any one of [1] to [7], wherein the sidewall of the sorting channel on the side connected to the orifice portion has at least one curvature. [9] The microchip according to [8], wherein the cross-sectional area of ​​the sorting channel increases continuously along the direction of the liquid flow up to a predetermined position.

[10] The microchip according to [9], wherein the sidewall of the sorting channel on the side connected to the orifice portion has two different curvatures.

[11] The macrochip according to

[10] , wherein the depth of the sorting channel is constant up to the second curvature portion, and the width of the sorting channel increases continuously along the direction of the liquid flow up to the two curvature portions.

[12] The microchip according to

[11] , wherein the depth of the sorting channel increases continuously in the direction of the liquid flow from the second curvature portion onwards until a predetermined position.

[13] the sorting channel and the orifice portion are formed in stacked substrate layers; The microchip according to any one of [8] to

[12] , wherein a part of the sorting channel and / or a part of the orifice portion is formed in a layer on one side of the substrate layer.

[14] The microchip according to any one of [1] to

[13] , wherein at least a portion of one surface of the substrate layer on which the sorting channels are formed is exposed to the outside.

[15] the main channel has a first optical detection region; The microchip according to any one of [1] to

[14] , wherein both surfaces of the substrate layer on which the first optical detection region is formed are exposed to the outside.

[16] the sorting channel has a second optical detection region; The microchip according to any one of [1] to

[15] , wherein both surfaces of the substrate layer on which the second optical detection region is formed are exposed to the outside.

[17] a sample liquid storage section in which the sample liquid is stored; a plate-like microchip having a sample liquid inlet into which a sample liquid is introduced, a main flow path through which the sample liquid introduced from the sample liquid inlet flows, and a fractionation flow path through which a target sample is fractionated from the sample liquid, the sample liquid inlet and the fractionation flow path having ends formed on the same side; and A sample collection kit in which the sample liquid storage section and the microchip are connected.

[18] A microparticle sorting device having a sample liquid inlet into which a sample liquid is introduced, a main flow path through which the sample liquid introduced from the sample liquid inlet flows, and a sorting flow path through which a target sample is sorted from the sample liquid, the device being equipped with a plate-shaped microchip on which the ends of the sample liquid inlet and the sorting flow path are formed on the same side.

[19] a chip insertion section into which the microchip is inserted; a light irradiation unit that irradiates light onto microparticles flowing through the main flow path; a light detection unit that detects scattered light and / or fluorescence emitted from the microparticles; a control unit that controls the direction of travel of microparticles flowing through the main flow path based on data detected by the light detection unit; The microparticle sorting device according to

[18] ,

[20] The microparticle sorting device according to

[18] or

[19] , further comprising a sample sorting kit including a sample liquid storage section in which the sample liquid is stored and a sample liquid storage section connected to the microchip, and further comprising a sample liquid delivery mechanism that delivers the sample from the sample liquid storage section to the microchip. [Explanation of symbols]

[0116] 100 microchips 101 Sample liquid inlet 102 sample liquid flow path 1021 Rapidly expanding section 103 Sheath fluid inlet 104 sheath fluid flow path 105 Main channel 106 First optical detection region 107 Particle sorting section 108 Branch Channel 1081 End of branch channel 108 109 Preparative flow path 1091 End of the fractionation channel 109 1092 Excitation area 1093 Second optical detection area 110 Buffer solution flow path 1101 Buffer solution inlet 111 Junction 120,130 Orifice section 150 Protection Department 151 Convex part d Width of the confluence channel T1~T5 Flow path connection parts 200 Sample Preparation Kit 201 Sample liquid storage section 2011 Pre-sample storage area 202 Filter section 203 Target sample storage section 204 Disposal Department 205 Sheath fluid storage section 206 Buffer solution storage section 207 Damper 208 Pressure Gauge Sensor 300 Microparticle separation device 301 Tip insertion part 302 Light irradiation unit 303 Light detection unit 304 Control Unit 305 Sample delivery mechanism

Claims

1. a sample liquid inlet into which the sample liquid is introduced; a main flow path through which the sample liquid introduced from the sample liquid inlet flows; a separation flow path through which a target sample is separated from the sample liquid; a sheath fluid inlet into which sheath fluid is introduced; A microchip having The sample liquid inlet, the end of the sorting channel, and the sheath liquid inlet are formed on the same side of the interior of the microchip in the longitudinal direction of the microchip.

2. A microchip as described in claim 1, wherein the sample liquid inlet, the end of the separation flow path, and the sheath liquid inlet are capable of communicating with their respective flow path connecting members, and the communicating portions with the respective flow path connecting members are formed on the same side in the longitudinal direction of the microchip.

3. Further, the liquid supply system has a buffer liquid inlet into which a buffer liquid is introduced; 2. The microchip according to claim 1, wherein the buffer solution inlets are formed on the same side of the microchip in the longitudinal direction thereof.

4. a branch channel branching from the main channel and through which samples other than the target sample are discarded; 2. The microchip according to claim 1, wherein ends of the branch channels are formed on the same side in the longitudinal direction of the microchip within the microchip.

5. A microchip as described in claim 3, wherein the buffer liquid inlet is capable of communicating with a flow path connecting member, and the communicating portion with the flow path connecting member is formed on the same side in the longitudinal direction of the microchip.

6. A microchip as described in claim 4, wherein the ends of the branched flow paths are connectable to a flow path connecting member, and the connecting portions with the flow path connecting member are formed on the same side in the longitudinal direction of the microchip.

7. A microchip as described in claim 2, having a protective portion that protects the flow path connection member.

8. 3. The microchip according to claim 2, wherein the main channel through which the sample liquid flows has, at an end on the sample liquid inlet side, a suddenly expanded portion having a cross-sectional area larger than the cross-sectional area of ​​the inner diameter of the channel connecting member.

9. an orifice portion that is coaxial with the main channel and is connected to the sorting channel; The microchip according to claim 1 , wherein a sidewall of the sorting channel on the side connected to the orifice portion has at least one curvature.

10. 10. The microchip according to claim 9, wherein the cross-sectional area of ​​the sorting channel increases continuously along the direction of the liquid flow up to a predetermined position.

11. The microchip according to claim 10 , wherein a side wall of the sorting channel on the side connected to the orifice portion has two different curvatures.

12. The macrochip according to claim 11 , wherein the sorting channel has a constant depth up to a second curvature portion, and a width that increases continuously along the direction of liquid flow up to the second curvature portion.

13. The microchip according to claim 12 , wherein the depth of the sorting channel increases continuously from the second curvature portion along the direction of the liquid flow.

14. the sorting channel and the orifice portion are formed in stacked substrate layers; The microchip according to claim 9 , wherein a part of the sorting channel and / or a part of the orifice portion is formed in a layer on one side of the substrate layer.

15. The microchip according to claim 14 , wherein at least a portion of one surface of the substrate layer on which the sorting channels are formed is exposed to the outside.

16. the main channel has a first optical detection region; The microchip according to claim 14 , wherein both surfaces of the substrate layer on which the first optical detection region is formed are exposed to the outside.

17. the sorting channel has a second optical detection region; The microchip according to claim 14 , wherein both surfaces of the substrate layer on which the second optical detection region is formed are exposed to the outside.

18. a sample liquid storage section in which the sample liquid is stored; a microchip having a sample liquid inlet into which a sample liquid is introduced, a main flow path through which the sample liquid introduced from the sample liquid inlet flows, a separation flow path through which a target sample is separated from the sample liquid, and a sheath liquid inlet into which a sheath liquid is introduced, wherein the sample liquid inlet, an end of the separation flow path, and the sheath liquid inlet are formed on the same side of the microchip in the longitudinal direction of the microchip; and A sample collection kit in which the sample liquid storage section and the microchip are connected.

19. A sample collection kit as described in Claim 18, wherein the microchip has the sample liquid inlet, the end of the collection flow path, and the sheath liquid inlet that are capable of communicating with their respective flow path connecting members, and the communicating portions with the respective flow path connecting members are formed on the same side in the longitudinal direction of the microchip.

20. A sample collection kit as described in claim 18, having a plurality of the microchips.

21. A microparticle sorting device equipped with a microchip having a sample liquid inlet into which a sample liquid is introduced, a main flow path through which the sample liquid introduced from the sample liquid inlet flows, a separation flow path through which a target sample is separated from the sample liquid, and a sheath liquid inlet into which a sheath liquid is introduced, wherein the sample liquid inlet, the end of the separation flow path, and the sheath liquid inlet are formed on the same side of the microchip in the longitudinal direction of the microchip.

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