JP7545729B2 - Cell sorters and flow cells - Google Patents

Description

The present invention relates to a cell sorter and a flow cell.

A cell sorter that separates cells is known (for example, Patent Document 1). A cell sorter is a device that analyzes each cell dispersed in a fluid using a fluorescent labeling technique, scattered light, etc., and further separates the target cells based on the information. Several methods have been reported as mechanisms for cell separation in cell sorters. For example, the cell sorter described in Patent Document 1 has a first flow path that connects a supply port that supplies a fluid containing cells and a discharge port that discharges the fluid, one or more second flow paths that intersect and connect with the first flow path, and a jet generating means that supplies a pulsed jet from one side of the second flow path and pushes cells that reach the intersection of the first flow path and the second flow path to the other side of the second flow path.

Japanese Patent Application Publication No. 1-170853

As a cell sorting mechanism in a cell sorter, a mechanism for sorting target cells by supplying a pulsed jet to the target cells has been known. In addition, as a method for forming a flow cell in a cell sorter, a soft lithography technique for forming a fine flow channel using a soft resin material such as PDMS (PolyDiMethylSiloxane) may be used. In forming a flow cell using such a soft resin, a structure in which multiple pillars (cylinders or square pillars) are provided to support the walls forming the flow channel and the chamber may be adopted in order to prevent deformation or collapse of the flow channel (particularly the part related to the cell sorting mechanism) and the chamber and maintain sufficient strength. However, such multiple pillars become obstacles to the fluid flowing in the flow channel when filling with liquid, which may disturb the flow of the fluid in the flow channel and generate air bubbles in the flow channel. In particular, in the cell sorting section related to cell sorting, if a pillar is provided in the flow channel or chamber that supplies a pulsed jet for cell sorting, the mixed air bubbles will cause the pressure of the jet to become unstable, and the accuracy of cell sorting will decrease. Therefore, there is room for improvement in terms of improving the accuracy of cell sorting while suppressing deformation and collapse of the flow channel and stabilizing the shape of the flow channel. Soft lithography methods using flexible resin materials such as PDMS have the advantage of being inexpensive and easy to mold, but when producing flow cells with wide (wide and low) flow channels or chamber structures, it is particularly necessary to install spacers such as supports to prevent the PDMS upper surface of the flow channel or chamber from collapsing, and structures with supports have the problem that gas is likely to remain where the supports are installed when filling with liquid, making it difficult to perform stable sorting due to the air bubbles that have been mixed in.

The present invention aims to provide a cell sorter and flow cell that can improve the accuracy of cell sorting by suppressing deformation and collapse of the flow path and stabilizing the flow of the circulating flow-changing fluid.

In order to achieve the above-mentioned object, the cell sorter of the present invention comprises a sample flow path through which a sample fluid containing cells flows, a flow transformation fluid container provided between a pair of opposing wall portions, connected to the sample flow path, and having a flow transformation fluid therein, a separation flow path arranged downstream of the flow transformation fluid container in the flow direction of the sample fluid and connected to the sample flow path, and a pressure transformation section which changes the liquid pressure inside the flow transformation fluid container and circulates the flow transformation fluid in a direction intersecting the flow direction of the sample fluid, wherein the flow transformation fluid container connects the pair of wall portions in a direction intersecting the flow direction of the sample fluid and the flow direction of the flow transformation fluid , and has a support plate extending in the flow direction of the flow transformation fluid , the support plate forming a plurality of parallel flow paths through which the flow transformation fluid flows, and the plurality of parallel flow paths each have equal flow path resistance to the flow of the flow transformation fluid.

In the above configuration, the parallel flow paths may extend parallel to the flow direction of the flow-changing fluid.

In the above configuration, the parallel flow paths may each have approximately the same length, height, and width.

In the above configuration, a chamber may be provided in which the pressure transformer is disposed, and the support plate may extend to a position along the outer shape of the chamber at the end on the chamber side.

In the above configuration, the flow transformation fluid storage section has a tip portion that communicates with the sample flow path, and the support plate may be installed so that the ratio of width to height does not exceed 10 in the tip flow path formed between the tip of the support plate opposite the pressure transformation section and the side wall of the tip portion.

In the above configuration, the width of the parallel flow passage may be greater than the thickness of the support plate.

In order to achieve the above-mentioned object, the flow cell of the present invention comprises a sample flow path through which a sample fluid containing cells flows, a flow transformation fluid container provided between a pair of opposing wall portions, connected to the sample flow path, and having a flow transformation fluid therein, and a separation flow path arranged downstream of the flow transformation fluid container in the flow direction of the sample fluid and connected to the sample flow path, wherein the flow transformation fluid container connects the pair of wall portions in a direction intersecting the flow direction of the sample fluid and the flow direction of the flow transformation fluid , and has a support plate extending in the flow direction of the flow transformation fluid , the support plate forming a plurality of parallel flow paths through which the flow transformation fluid flows, and the plurality of parallel flow paths each having equal flow path resistance to the flow of the flow transformation fluid.

The present invention provides a cell sorter and flow cell that can improve the accuracy of cell sorting by suppressing deformation and collapse of the flow path and stabilizing the flow of the circulating flow-changing fluid.

FIG. 1 is a perspective view of a cell sorter according to one embodiment. FIG. 1 is a plan view of a cell sorter according to one embodiment. FIG. 2 is an enlarged perspective view of the vicinity of a main body of a flow transformation fluid container according to one embodiment.

(Cell sorter)
Hereinafter, a cell sorter according to an embodiment of the present invention will be described with reference to the drawings.
FIG. 1 is a perspective view of a cell sorter 1 according to one embodiment.
The cell sorter 1 of this embodiment is used to separate target cells from a sample fluid A containing multiple types of cells. The target cells are identified based on scattered light or fluorescent data obtained by irradiating multiple types of cells contained in the sample fluid A with, for example, laser light. Machine learning can be used to identify the target cells. In this case, a discrimination model is created from teacher information obtained from learning cells measured in advance, and the target cells are identified based on the discrimination model.
As shown in FIG. 1, the cell sorter 1 includes a flow cell 10, a cell information acquisition device 2, and a piezoelectric element 3 (corresponding to a transformer in the claims).

(Flow cell)
The flow cell 10 is a rectangular plate-like member extending in one direction, and is formed, for example, by bonding together a rectangular plate-like first member 4 and a rectangular plate-like second member 5.
The first member 4 is formed of, for example, glass or PDMS (PolyDiMethylSiloxane).
The second member 5 may be the same as the first member 4, or may be different, and is formed of a transparent and flexible resin material such as PDMS (PolyDiMethylSiloxane). A sample flow path 11, a sample fluid supply unit 20, a sheath flow path 12, a sheath liquid supply unit 13, a flow transformation fluid storage unit 14, and a separation flow path 15 are formed on the first member 4 side of the second member 5. The sample flow path 11, the sample fluid supply unit 20, the sheath flow path 12, the sheath liquid supply unit 13, the flow transformation fluid storage unit 14, and the separation flow path 15 are covered by the first member 4.

The sample flow channel 11 extends in the longitudinal direction of the flow cell 10. Sample fluid A containing cells flows through the sample flow channel 11 along the longitudinal direction of the flow cell 10. One end of the sample flow channel 11 communicates with a sample fluid supply unit 20. The other end of the sample flow channel 11 communicates with a fractionation flow channel 15. The sample fluid A is supplied from the sample fluid supply unit 20 and flows through the sample flow channel 11 from one end to the other end.
Hereinafter, the flow direction of the sample fluid A is referred to as a flow direction D1.
The sample flow path 11 has an upstream end 11a in the flow direction D1, a throttle flow path 11b extending along the flow direction D1, a junction 11c provided at the downstream end of the throttle flow path 11b, an alignment flow path 11d extending downstream from the junction 11c along the flow direction D1, a cell sorting section 11e provided at the downstream end of the alignment flow path 11d, and a discharge flow path 11f extending downstream from the cell sorting section 11e along the flow direction D1.

A sample fluid A is supplied to the upstream end 11a.
The throttle flow passage 11b is provided at its downstream end with a throttle section 11g. In the throttle flow passage 11b, the section from the upstream end 11a to the throttle section 11g has the same flow passage width. In the throttle section 11g, the flow passage width becomes narrower toward the downstream side.
The confluence 11 c connects the sample channel 11 and the sheath channel 12 .
The alignment channel 11d aligns the cells in the sample fluid A in a line along the flow direction D1.
The cell sorting section 11e sorts out target cells to be sorted out from among the cells aligned in a line in the alignment channel 11d.
The sample fluid A that has passed through the cell sorting section 11e flows through the discharge flow path 11f. The sample fluid A that has passed through the discharge flow path 11f is discharged into a test tube (not shown) or the like that is disposed downstream of the downstream end of the discharge flow path 11f.

The sample fluid supplying unit 20 is connected to the upstream end 11a in the flow direction D1 of the sample flow path 11. The sample fluid supplying unit 20 supplies a sample fluid A to the sample flow path 11. The sample fluid supplying unit 20 shown in Fig. 1 includes two sample openings 21 and a communication passage 22 that connects the two sample openings 21 with the upstream end 11a.
Of the two sample openings 21, the one arranged on the upstream side in the flow direction D1 is referred to as a first sample opening 21a. Of the two sample openings 21, the one arranged on the downstream side in the flow direction D1 is referred to as a second sample opening 21b, and a first tube 23a is connected to the first sample opening 21a, and a second tube 23b is connected to the second sample opening 21b. The sample fluid A is supplied to the sample fluid supply unit 20 through the first tube 23a or the second tube 23b.
The communication passage 22 extends along the flow direction D1. The first sample opening 21a is provided at an upstream end of the communication passage 22 in the flow direction D1. The second sample opening 21b is provided at a downstream end of the communication passage 22 in the flow direction D1.

The sheath flow channel 12 is formed alongside the sample flow channel 11. An example in which two sheath flow channels 12 are formed is shown in Fig. 1, but the number is not limited to two. In Fig. 1, the two sheath flow channels 12 are formed symmetrically with the sample flow channel 11 in between. Sheath liquid B flows through the sheath flow channel 12. The sheath liquid B aligns the cells in a row in the alignment flow channel 11d and flows continuously through the alignment flow channel 11d.
The sheath liquid B flows through the sheath flow channel 12 in the same direction as the sample fluid A, that is, from the upstream side to the downstream side of the flow direction D1 of the sample fluid A.
The two sheath flow paths 12 are in communication with each other at their upstream ends 12a and at their downstream ends 12b in the flow direction of the sheath fluid B.
The downstream ends 12 b of the two sheath channels 12 communicate with a junction 11 c of the sample channel 11 .
The sheath fluid supply unit 13 is provided at the upstream ends 12a of the two sheath flow paths 12. The sheath fluid supply unit 13 is in communication with the upstream ends 12a of the two sheath flow paths 12. The sheath fluid supply unit 13 supplies sheath fluid B to the two sheath flow paths 12.

FIG. 2 is a plan view of the cell sorter 1 according to one embodiment.
FIG. 3 is an enlarged perspective view of the vicinity of the main body 30 of the flow diversion fluid container 14 according to one embodiment.
As shown in FIG. 2, a pair of flow-changing fluid containers 14 are arranged on either side of the sample flow path 11. The flow-changing fluid containers 14 are formed symmetrically on either side of the sample flow path 11. The flow-changing fluid containers 14 are provided between the wall 4a of the first member 4 constituting the flow cell 10 and the wall 5a of the second member 5 constituting the flow cell 10 (see FIG. 3). That is, the flow-changing fluid containers 14 are provided between a pair of opposing walls 4a and 5a. The flow-changing fluid containers 14 communicate with the cell sorting section 11e of the sample flow path 11. A flow-changing fluid E is contained inside the flow-changing fluid containers 14. The flow-changing fluid E is a fluid that is circulated for sorting the target cells.
The flow transformation fluid accommodation portion 14 has a main body portion 30 and a support plate 33 .

The main body 30 extends in a direction perpendicular to the flow direction D1 of the sample fluid A. The main body 30 communicates with the cell sorting section 11e of the sample flow path 11. The main body 30 has a tip 30a communicating with the cell sorting section 11e and a parallel section 30b extending from the tip 30a in a direction perpendicular to the flow direction D1.
The tip portion 30a becomes wider as it moves away from the cell sorting portion 11e in a direction perpendicular to the flow direction D1 of the sample fluid A. The parallel portion 30b extends with a constant width in the direction perpendicular to the flow direction D1. The width of the parallel portion 30b is, for example, 5.8 mm.
The chamber 31 is provided at a position farther away from the cell sorting section 11e than the main body section 30. The chamber 31 penetrates the second member 5 in the thickness direction. The chamber 31 is formed in a circular shape in a plan view. The diameter of the chamber 31 is, for example, 8 mm. The penetration surface of one of the pair of chambers 31 on the opposite side to the first member 4 is covered by a piezoelectric element 3. Therefore, the diameter of the chamber 31 is formed to be smaller than the diameter of the piezoelectric element 3. The other of the pair of chambers 31 is covered by, for example, a transparent glass plate 6. Note that, in the figure, an example in which the piezoelectric element 3 is installed on one side of the pair of chambers 31 is described, but the piezoelectric element 3 may be installed on both of the pair of chambers 31.

The flow change fluid discharge path 32 extends from the chamber 31 toward the upstream side in the flow direction D1. The flow change fluid discharge path 32 has a flow change fluid discharge section (not shown) at the end opposite to the chamber 31. The flow change fluid E is supplied from the sheath liquid supply section 13, flows through the sample flow path 11, passes through the cell sorting section 11e through the parallel flow path 34 described later, fills the chamber 31, and is discharged from the flow change fluid discharge path 32 (when filling with liquid). The flow change fluid discharge path 32 is closed except when filling with liquid. The flow change fluid E flows in the main body 30 from the piezoelectric element 3 side toward the sample flow path 11 side so as to be perpendicular to the flow direction D1 of the sample fluid A.
Hereinafter, the flow direction of the flow diversion fluid E in the main body 30 is referred to as a flow direction D2.

In the examples of Figs. 1 to 3, five support plates 33 are provided inside the main body 30. The support plates 33 are rectangular plate-shaped members. The five support plates 33 are formed so that the cross sections in the thickness direction of the first member 4 and the second member 5 have the same shape. The support plates 33 extend parallel to the flow direction D2 of the flow conversion fluid E. The height direction of the support plates 33 is along the thickness direction of the first member 4 and the second member 5. The thickness d of the support plates 33 is, for example, 200 µm. The support plates 33 connect the wall 4a of the first member 4 and the wall 5a of the second member 5. Therefore, the height of the parallel flow passage 34 is equal to the height of the support plates 33, and in the examples of these figures, the height of the parallel flow passage 34 is 200 µm. The support plates 33 extend to a position along the outer shape of the chamber 31 at the end on the chamber 31 side where the piezoelectric element 3 is arranged. The support plates 33 are arranged at equal intervals in the flow direction D1 of the sample fluid A. Six parallel flow paths 34 through which the flow diversion fluid E flows are formed along the five support plates 33. Specifically, four parallel flow paths 34 are formed between adjacent support plates 33, and two parallel flow paths 34 are formed between the support plates 33 and the side wall of the parallel portion 30b.
The six parallel flow paths 34 extend in parallel to the flow direction D2 of the flow transformation fluid E. The six parallel flow paths 34 are formed so that each has the same flow path resistance (hydrodynamic resistance) to the flow of the flow transformation fluid E. The flow path resistance is determined by the length, height, and width of each flow path, and in the examples of Figures 1 to 3, the six parallel flow paths 34 are formed to have approximately the same length, height, and width, so that each of the six parallel flow paths 34 has the same flow path resistance to the flow of the flow transformation fluid E.
The parallel flow path 34 and the support plate 33 forming it are formed so that the tip on the cell sorting section 11e side is sufficiently close to the cell sorting section 11e. In the tip flow path 35 formed in the part sandwiched between the tip of the support plate 33 opposite to the piezoelectric element 3 and the side wall 30a1 of the tip 30a on the cell sorting section 11e side of the flow conversion fluid storage section 14, the ratio of the width w2 of the tip flow path 35 to the height h (height h of the flow conversion fluid storage section) of the tip flow path 35 is formed so as not to exceed 10. Also, in the tip flow path 35, the ratio of the width w2 to the height h is greater than 1. The width w1 of the parallel flow path 34 is, for example, 0.8 mm in the parallel portion 30b. When the height of the parallel flow path 34 (height of the flow conversion fluid storage section 14) is h, the parallel flow path 34 is formed so as to satisfy w1/h<10. With this structure, it is possible to suppress the collapse of the flow path due to the bending of the first member 4 and the second member 5.
The width w 1 of the parallel flow passage 34 is greater than the thickness d of the support plate 33 .

As shown in FIG. 2, the sorting channel 15 is disposed downstream of the flow conversion fluid storage section 14 in the flow direction D1 of the sample fluid A. The sorting channel 15 is connected to the cell sorting section 11e of the sample channel 11. A pair of sorting channels 15 are provided on either side of the sample channel 11. The sorting channels 15 are formed symmetrically on either side of the sample channel 11. The sorting channel 15 extends in the flow direction D1 along the discharge channel 11f of the sample channel 11. A test tube (not shown) or the like for collecting the sorted target cells is disposed downstream of the downstream end of the sorting channel 15.

(Cell information acquisition device)
As shown in FIG. 1, the cell information acquisition device 2 is connected to the alignment channel 11d of the sample channel 11. The cell information acquisition device 2 includes, for example, a laser light source (not shown) and a detector (not shown). The cell information acquisition device 2 is equipped with a control unit (not shown) and the like. The cell information acquisition device 2 irradiates a laser beam onto cells contained in the sample fluid A. The cell information acquisition device 2 detects the amount of ions generated from the cells by the irradiation of the laser beam. The scattered light and the fluorescent light are detected by a detector to obtain information on the morphology of the cells and the internal structure of the cells, such as nuclei and granules. The control unit discriminates target cells to be separated from the cells. The cell information acquisition device 2 according to this embodiment may have a function of learning the characteristics of the target cells by machine learning.

(Piezoelectric element)
The piezoelectric element 3 is formed in a disk shape. The piezoelectric element 3 is disposed so that the center of the piezoelectric element 3 coincides with the center of the chamber 31, and is attached in a manner that covers the through-hole surface of the chamber 31 on the opposite side to the first member 4. Therefore, the outer periphery of the chamber 31 is smaller than the outer periphery of the piezoelectric element 3. The piezoelectric element 3 is fixed by applying an adhesive or the like to the surface side that is not in contact with the second member 5 so as to straddle the piezoelectric element 3 and the second member 5 in a plan view. Therefore, the adhesive or the like that fixes the piezoelectric element 3 does not penetrate into the chamber 31. The ends of the five support plates 33 on the chamber 31 side are provided so as to follow the outer shape of the chamber 31 in a plan view.
The piezoelectric element 3 changes the liquid pressure inside the flow transformation fluid storage section 14, and causes the flow transformation fluid E to flow in a direction (flow direction D2) intersecting the flow direction D1 of the sample fluid A. The piezoelectric element 3 is electrically connected to the cell information acquisition device 2. A pulse-shaped voltage, for example, is applied to the piezoelectric element 3 from the cell information acquisition device 2. The piezoelectric element 3 deforms in response to the applied voltage. The liquid pressure inside the flow transformation fluid storage section 14 changes due to the deformation of the piezoelectric element 3. The flow transformation fluid E is caused to flow in the flow direction D2 due to the change in liquid pressure inside the flow transformation fluid storage section 14 caused by the piezoelectric element 3.

(How to use a cell sorter)
An example of a method of using the cell sorter 1 will now be described with reference to FIG.
The method of using the cell sorter 1 includes an equipment adjustment step, a learning step, and a fractionation step. During use of the cell sorter 1, a sample fluid A containing a plurality of samples flows through the flow cell 10. The sample fluid A includes an equipment adjustment sample fluid A1 containing standard beads that is circulated in the equipment adjustment step, a learning sample fluid A2 containing a learning sample that is circulated in the learning step, and a fractionation sample A3 containing a fractionation sample that is circulated in the fractionation step.
(Equipment adjustment process)
In the equipment adjustment step, a sample fluid A1 for equipment adjustment containing a sample for equipment adjustment is supplied from the first sample opening 21a of the sample fluid supply unit 20. The equipment adjustment step is a step for checking whether the cell sorter 1 to be used is in a state suitable for measurement and, if necessary, for adjusting it in advance, and the sample fluid A1 for equipment adjustment contains, for example, standard beads whose particle characteristics are known in advance. The sample fluid A1 for equipment adjustment flows through the sample fluid supply unit 20 and the sample flow path 11. The particles contained in the sample fluid A1 for equipment adjustment flow in a single row in the alignment flow path 11d of the sample flow path 11. Next, information on each particle flowing through the alignment flow path 11d is acquired by the cell information acquisition device 2, and the operating state of the cell sorter 1 can be confirmed.
(Learning process)
In the learning step, first, a learning sample fluid A2 containing a learning sample is supplied from the first sample opening 21a of the sample fluid supply unit 20. The learning sample fluid A2 contains, for example, a plurality of types of cells including a target cell to be separated. The target cells contained in the learning sample fluid A2 flow in a single row in the alignment channel 11d of the sample channel 11.
Next, the cell information acquisition device 2 reads information from each target cell flowing through the alignment channel 11d and learns criteria for discriminating the target cells. That is, the cell information acquisition device 2 creates a discrimination model for discriminating the target cells based on the information of each learning sample A2 flowing through the alignment channel 11d.

(Isolation process)
In the sorting step, first, a sorting sample fluid A3 is supplied from the second sample opening 21b of the sample fluid supply unit 20. The sorting sample fluid A3 contains a plurality of types of cells including the target cells to be sorted. The plurality of cells contained in the sorting sample fluid A3 are aligned in a single row and flow through the alignment channel 11d of the sample channel 11.
Next, the cell information acquisition device 2 distinguishes the target cell from the various types of cells flowing through the alignment flow path 11d based on the discrimination criteria for the target cell learned in the learning process. When the cell information acquisition device 2 distinguishes the target cell, it applies, for example, a pulsed voltage to the piezoelectric element 3.

The piezoelectric element 3 is deformed when, for example, a pulsed voltage is applied from the cell information acquisition device 2. The flow transformation fluid container 14 on one side in which the piezoelectric element 3 is installed is depressurized by the deformation of the piezoelectric element 3. When the liquid pressure of the flow transformation fluid container 14 is depressurized, the flow transformation fluid E flows along a flow direction D2 perpendicular to the flow direction D1.
The flow conversion fluid E flows from the cell sorting section 11e toward the chamber 31, passing through the six parallel flow paths 34. The flow of the flow conversion fluid E draws the target cells that have reached the cell sorting section 11e into the flow conversion fluid storage section 14 on the piezoelectric element 3 side, across the sample flow path 11. The target cells are drawn in by the flow pressure, and the direction of movement changes from the flow direction D1 to the direction along the sorting flow path 15. The target cells move to the sorting flow path 15 on the piezoelectric element 3 side, across the sample flow path 11, out of the pair of sorting flow paths 15.

The target cells that have moved to the sorting channel 15 move into a test tube placed at the downstream end of the sorting channel 15 .
Through the above steps, the target cells are separated and isolated.
In this embodiment, the liquid pressure in the flow transformation fluid storage section 14 is reduced by the piezoelectric element 3, but it may be increased. In this case, the flow transformation fluid E flows in the opposite direction to when the pressure is reduced by the piezoelectric element 3, and the target cells are moved to the sorting flow path 15 on the opposite side of the sample flow path 11 to the piezoelectric element 3.

(Action, Effect)
According to the above-described embodiment, the following actions and effects can be obtained.
The flow conversion fluid storage section 14 has a support plate 33 that connects the wall 4a of the pair of first members 4 and the wall 5a of the pair of second members 5. As a result, the support plate 33 bears the weight of the wall 5a, and can support the wall 5a of the flow conversion fluid storage section 14. In other words, the support plate 33 supports the wall 5a and maintains the structure of the wall 5a of the flow conversion fluid storage section 14. Therefore, deformation and collapse of the flow conversion fluid storage section 14, which serves as a flow path through which the flow conversion fluid E flows during cell sorting, can be suppressed, and the shape of the parallel section flow path 34 formed therein can be stabilized.
The parallel flow paths 34 have equal flow path resistances to the flow of the flow conversion fluid E. As a result, when filling the liquid, the flow conversion fluid E that reaches the end of the parallel flow path 34 from the cell sorting section 11e flows evenly into the parallel flow paths 34, reaches the end of the chamber 31, and merges again. This makes it possible to suppress irregular fluctuations in the flow rate and direction of the flow conversion fluid E flowing through the flow conversion fluid storage section 14, thereby suppressing the generation of air bubbles in the flow conversion fluid storage section 14. Therefore, the flow pressure of the flow conversion fluid E can be stabilized during cell sorting, so that the cells in the sample fluid A can be stably moved to the sorting flow path 15 by the flow pressure of the flow conversion fluid E. This improves the accuracy of cell sorting.
By providing multiple parallel flow paths 34, the overall flow path resistance of the flow conversion fluid storage section 14 can be reduced. This makes it possible to provide the cell sorting section 11e with a flow rate sufficient for cell sorting. This allows the cells in the sample fluid A to stably move to the sorting flow path 15. This improves the accuracy of cell sorting.
Therefore, deformation and collapse of the flow-changing fluid container 14, which serves as a flow path through which the flow-changing fluid E flows during cell sorting, can be suppressed, and the accuracy of cell sorting can be improved while stabilizing the shape of the parallel flow path 34 formed therein.

Since the multiple parallel flow paths 34 extend in parallel to the flow direction D2 of the flow conversion fluid E, the multiple parallel flow paths 34 have equal flow path resistance to the flow of the flow conversion fluid E. As a result, when filling with liquid, the flow conversion fluid E that reaches the end of the parallel flow path 34 from the cell sorting section 11e flows evenly into the multiple parallel flow paths 34, reaches the end of the chamber 31 and merges again. This makes it possible to suppress irregular fluctuations in the flow speed and direction of the flow conversion fluid E flowing through the flow conversion fluid storage section 14, thereby suppressing the generation of air bubbles in the flow conversion fluid storage section 14. Therefore, since the flow pressure of the flow conversion fluid E can be stabilized during cell sorting, the cells in the sample fluid A can be stably moved to the sorting flow path 15 by the flow pressure of the flow conversion fluid E. This improves the accuracy of cell sorting.

The support plate 33 is provided so that multiple flow path structures are formed in parallel inside the flow conversion fluid storage section 14 (parallel flow paths 34). The flow path resistance of the parallel flow paths 34 to the flow of the flow conversion fluid E can be adjusted by the length, height, and width of the parallel flow paths 34. In the examples of Figures 1 to 3, the multiple parallel flow paths 34 have the same length, height, and width. As a result, the flow path resistance to the flow of the flow conversion fluid E can be made uniform among the multiple parallel flow paths 34, so the flow rate of the flow conversion fluid E can be made uniform among the multiple parallel flow paths 34. Therefore, irregular fluctuations in the flow speed and direction of the flow conversion fluid E in the parallel flow paths 34 during liquid filling can be suppressed, so that the generation of bubbles in the parallel flow paths 34 can be suppressed. Therefore, the flow pressure of the flow conversion fluid E can be stabilized, so that the cells in the sample fluid A can be stably moved to the separation flow path 15 by the flow pressure of the flow conversion fluid E. This improves the accuracy of cell separation.

The support plate 33 extends to a position along the outer shape of the chamber 31 at the end on the chamber 31 side. This makes it possible to suppress irregular fluctuations in the flow rate and direction of the flow transformation fluid E in the parallel section flow path 34 at the end on the chamber 31 side of the support plate 33 when the piezoelectric element 3 generates a flow of the flow transformation fluid E in the parallel section flow path 34, thereby suppressing the generation of air bubbles in the parallel section flow path 34. This makes it possible to stabilize the flow pressure of the flow transformation fluid E, so that the cells in the sample fluid A can be stably moved to the separation flow path 15 by the flow pressure of the flow transformation fluid E. This improves the accuracy of cell separation.

In the above configuration, the support plate 33 is installed so that the ratio of the width w2 to the height h in the tip flow passage 35 formed between the tip of the support plate 33 opposite the piezoelectric element 3 and the side wall 30a1 of the tip 30a does not exceed 10. As a result, the wall 5a at the position corresponding to the tip flow passage 35 is supported by the side wall 30a1 and the support plate 33, so that the first member 4 and the second member 5 can be prevented from bending in the tip flow passage 35. Therefore, the collapse of the tip flow passage 35 can be prevented.

In the tip flow passage 35, the ratio of the width w2 to the height h is greater than 1. This allows the flow conversion fluid E to flow evenly into the multiple parallel flow passages 34 via the tip flow passage 35.

The width w1 of the parallel flow passage 34 is greater than the thickness d of the support plate 33. This allows the parallel flow passage 34 to be formed wide while maintaining the strength of the parallel section 30b where the support plate 33 is arranged. Therefore, the flow of the flow conversion fluid E in the flow conversion fluid storage section 14 including the parallel flow passage 34 can be made uniform, while the flow resistance of the entire flow conversion fluid storage section 14 can be reduced.

Although the preferred embodiments of the present invention have been described above, the present invention is not limited to these embodiments. Addition, omission, substitution, and other modifications of the configuration are possible without departing from the spirit of the present invention. The present invention is not limited by the above description, but is limited only by the scope of the appended claims.

In addition, the components in the above-described embodiments may be replaced with well-known components as appropriate without departing from the spirit of the present invention.

1...cell sorter, 3...piezoelectric element (transformation section), 4a...wall section, 5a...wall section, 11...sample flow path, 14...fluid storage section for flow conversion, 15...separation flow path, 30a...tip section, 30a1...side wall, 31...chamber, 33...support plate, 34...parallel flow path, 35...tip flow path, A...sample fluid, D1...flow direction of sample fluid, D2...flow direction of flow conversion fluid, E...fluid for flow conversion, d...thickness of support plate, h...height of parallel flow path and tip flow path, w1...width of parallel flow path, w2...width of tip flow path

Claims (7)

a sample flow path through which a sample fluid containing cells flows;
a flow conversion fluid storage section provided between a pair of opposing walls, communicating with the sample flow path, and having a flow conversion fluid therein;
a separation flow path that is disposed downstream of the flow conversion fluid storage unit in a flow direction of the sample fluid and communicates with the sample flow path;
a pressure change unit that changes the liquid pressure inside the flow change fluid container and causes the flow change fluid to flow in a direction intersecting the flow direction of the sample fluid;
Equipped with
The flow transformation fluid storage portion connects the pair of walls in a direction intersecting the flow direction of the sample fluid and the flow direction of the flow transformation fluid , and has a support plate extending in the flow direction of the flow transformation fluid ,
The support plate forms a plurality of parallel flow paths through which the flow transformation fluid flows,
A cell sorter, wherein the parallel flow paths each have an equal flow resistance to the flow of the flow conversion fluid. The cell sorter according to claim 1, characterized in that the parallel flow paths extend parallel to the flow direction of the flow conversion fluid. The cell sorter according to claim 1 or 2, characterized in that the parallel flow paths are each approximately equal in length, height, and width. a chamber in which the transformer unit is disposed,
4. The cell sorter according to claim 1, wherein the support plate extends at an end on the chamber side to a position along the outer shape of the chamber. the flow conversion fluid container has a tip portion communicating with the sample flow path,
5. A cell sorter as claimed in any one of claims 1 to 4, characterized in that the support plate is installed so that the ratio of width to height in a tip flow path formed between the tip of the support plate opposite the transformer section and the side wall of the tip does not exceed 10. The cell sorter according to any one of claims 1 to 5, characterized in that the width of the parallel flow path is greater than the thickness of the support plate. a sample flow path through which a sample fluid containing cells flows;
a flow conversion fluid storage section provided between a pair of opposing walls, communicating with the sample flow path, and having a flow conversion fluid therein;
a separation flow path that is disposed downstream of the flow conversion fluid container in a flow direction of the sample fluid and communicates with the sample flow path;
Equipped with
The flow transformation fluid storage portion connects the pair of walls in a direction intersecting the flow direction of the sample fluid and the flow direction of the flow transformation fluid , and has a support plate extending in the flow direction of the flow transformation fluid ,
The support plate forms a plurality of parallel flow paths through which the flow transformation fluid flows,
A flow cell characterized in that the parallel flow paths each have an equal flow resistance to the flow of the flow conversion fluid.

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