This application is entitled to the benefit of Japanese Patent Application No. 2020-055818, filed on Mar. 26, 2020, the disclosure of which including the specification, drawings and abstract is incorporated herein by reference in its entirety.
The present invention relates to a liquid handling device and a liquid handling method for forming a sheath flow.
In recent years, channel chips have been used for the analysis of cells, proteins, nucleic acids, and other substances. The advantage of flow chips is that the amount of sample and reagents required for analysis can be reduced, and they are expected to be used in various applications such as clinical, food, and environmental testing.
For example, PTL 1 discloses a channel chip for sorting microparticles (e.g., cells) in a liquid flowing through a main channel and extracting only the desired microparticles. The channel chip disclosed in PTL 1 is manufactured by laminating three substrates in which predetermined grooves and through holes are formed. In this channel chip, two sheath liquid channels in which the sheath liquid flows are joined, from both sides, to the sample liquid channel in which the sample liquid containing microparticles flows. Thus, the sheath liquid joining the sample liquid from both sides forms a sheath flow in which the laminar flow of the sample liquid is sandwiched between the two laminar flows of the sheath liquid in the main flow channel located downstream of the confluence point.
PTL 1
Japanese Patent Application Laid-Open No. 2014-036604
The channel chip disclosed in PTL 1 is manufactured by laminating as many as three substrates, which results in a large manufacturing cost. In addition, the channel chip disclosed in PTL 1 forms a three-layered sheath flow in which the laminar flow of the sample liquid is sandwiched between two laminar flows of sheath liquid, but depending on the use, it may be desired to form a two-layered sheath flow in which the laminar flow of the sample liquid is surrounded by the laminar flow of sheath liquid.
An object of the present invention is to provide a liquid handling device including a substrate and a film joined to the substrate and can form the sheath flow in which the sheath liquid surrounds the main liquid. In addition, another object of the present invention is to provide a liquid handling method of forming the sheath flow using the liquid handling device.
A liquid handling device of an embodiment of the present invention is configured to form a sheath flow including main liquid and sheath liquid surrounding the main liquid, the liquid handling device including: a substrate; a film joined to the substrate; a main channel configured to carry the main liquid; a first sheath liquid channel that opens to a bottom surface of the main channel, the first sheath liquid channel being configured to carry the sheath liquid; and a second sheath liquid channel that opens to a top surface of the main channel, the second sheath liquid channel being configured to carry the sheath liquid. At least one of a confluence part of the first sheath liquid channel with the main channel and a confluence part of the second sheath liquid channel with the main channel is made up of the substrate and a curved portion of the film, the curved portion of the film being curved in a direction away from the substrate.
A liquid handling method of an embodiment of the present invention is a method of forming a sheath flow using the liquid handling device, the liquid handling method including: forming a sheath flow including the main liquid and the sheath liquid surrounding the main liquid by carrying the main liquid in the main channel and carrying the sheath liquid in the first sheath liquid channel and the second sheath liquid channel.
According to the present invention, the sheath flow in which the sheath liquid surrounds the main liquid can be formed using a simple device.
As illustrated in these drawings, liquid handling device 100 includes substrate 110 and film 120. In substrate 110, a groove configured to be a channel and a through hole configured to be an inlet or an outlet are formed. Film 120 is joined to one surface of substrate 110 to close the openings of the groove and the through hole formed in substrate 110. A part of the region of film 120 is formed as curved portion 121 curved in a direction away from substrate 110. The groove of substrate 110 closed with film 120 serves as a channel for carrying main liquid or sheath liquid. In addition, the inner space of curved portion 121 also serves as a channel for carrying sheath liquid.
The thickness of substrate 110 is not limited. For example, the thickness of substrate 110 is 1 mm to 10 mm. In addition, the material of substrate 110 is not limited.
For example, the material of substrate 110 is appropriately selected from publicly known resins and glass. Examples of the material of substrate 110 include polyethylene terephthalate, polycarbonate, polymethylmethacrylate, polyvinyl chloride, polypropylene, polyether, polyethylene, polystyrene, cycloolefin resin, silicone resin and elastomer.
The thickness of film 120 is not limited as long as a required strength can be ensured. For example, the thickness of film 120 is 30 μm to 300 μm. In addition, the material of film 120 is not limited as long as curved portion 121 can be molded. For example, the material of film 120 is appropriately selected from publicly known resins. Examples of the material of film 120 include polyethylene terephthalate, polycarbonate, polymethylmethacrylate, polyvinyl chloride, polypropylene, polyether, polyethylene, polystyrene, cycloolefin resin, silicone resin and elastomer. For observation of the main liquid flowing inside the channel, it is preferable that the material of film 120 be transparent. In addition, for fluorescence observation of the main liquid flowing inside the channel, it is preferable that the material of film 120 be a material that transmits excitation light and fluorescence, with low autofluorescence. Film 120 is joined to substrate 110 by, for example, thermal welding, laser welding, an adhesive agent and the like.
In the present embodiment, liquid handling device 100 includes main liquid introduction part 130, main channel 131, two first sheath liquid introduction parts 132, two first sheath liquid channels 133, two second sheath liquid introduction parts 134, two second sheath liquid channels 135, and liquid ejection part 136.
Main liquid introduction part 130 is a bottomed recess to which main liquid is introduced. In the present embodiment, main liquid introduction part 130 is composed of first through hole 111 formed in substrate 110, and a part of film 120 that closes one opening of first through hole 111 (see
The type of the main liquid that is introduced to main liquid introduction part 130 is not limited. For example, the main liquid is cell suspension, DNA-containing liquid, RNA-containing liquid, or the like. The main liquid may be liquid containing (sample liquid) a sample such as blood or an analyte such as cells and nucleic acids, liquid containing reagent, beads and the like, or a mixture of liquid containing an analyte and liquid containing reagent, beads and the like.
Main channel 131 is a channel in which liquid can move. The upstream end of main channel 131 is connected to main liquid introduction part 130, and the downstream end of main channel 131 is connected to liquid ejection part 136. In addition, as described later, first sheath liquid channel 133 and second sheath liquid channel 135 are connected to main channel 131 at respective positions different from each other. In the present embodiment, the main liquid is introduced from main liquid introduction part 130 to main channel 131, and the sheath liquid is introduced from the confluence part with first sheath liquid channel 133 and the confluence part with second sheath liquid channel 135. Accordingly, the main liquid flows in main channel 131 in the region from main liquid introduction part 130 to the confluence part with first sheath liquid channel 133, and the main liquid and the sheath liquid flow in main channel 131 in the region from the confluence part with first sheath liquid channel 133 to liquid ejection part 136. As described later, in the region from the confluence part with second sheath liquid channel 135 to liquid ejection part 136, the main liquid and the sheath liquid flow in main channel 131 in a sheath flow state in which the sheath liquid surrounds the main liquid.
Main channel 131 is composed of first groove 112 formed in substrate 110 and a part of film 120 that closes the opening of first groove 112 (see
Two first sheath liquid introduction parts 132 and two second sheath liquid introduction parts 134 are bottomed recesses to which the sheath liquid is introduced. In the present embodiment, each of two first sheath liquid introduction parts 132 is composed of second through hole 113 formed in substrate 110 and a part of film 120 that closes one opening of second through hole 113 (see
The type of the sheath liquid introduced to first sheath liquid introduction part 132 and second sheath liquid introduction part 134 is not limited. For example, the sheath liquid is physiological saline, buffer solution, or the like. For fluorescence observation of the main liquid flowing inside the channel, it is preferable that the sheath liquid be liquid with low optical interference.
Two first sheath liquid channels 133 are channels in which liquid can move. The upstream ends of two first sheath liquid channels 133 are connected to respective first sheath liquid introduction parts 132 different from each other, and the downstream ends of two first sheath liquid channels 133 are connected to main channel 131. In first sheath liquid channel 133, the sheath liquid introduced to first sheath liquid introduction part 132 flows. First sheath liquid channel 133 opens at least to the bottom surface of main channel 131. In the present embodiment, the “bottom surface of a channel” means a surface opposite to film 120 in the inner surfaces of the channel In the present embodiment, two first sheath liquid channels 133 open not only to the bottom surface of main channel 131 but also to side surfaces of main channel 131 opposing each other (see
Each of two first sheath liquid channels 133 is composed of second groove 114 formed in substrate 110 and a part of film 120 that closes the opening of second groove 114 (see
Two second sheath liquid channels 135 are channels in which liquid can move. The upstream ends of two second sheath liquid channels 135 are connected to respective second sheath liquid introduction parts 134 different from each other, and the downstream ends of two second sheath liquid channels 135 are connected to main channel 131. The sheath liquid introduced to second sheath liquid introduction part 134 flows in second sheath liquid channel 135. Second sheath liquid channel 135 opens at least to the top surface of main channel 131. In the present embodiment, the “top surface of a channel” means a surface on film 120 side (the surface formed with film 120) of the inner surfaces of the channel In the present embodiment, two second sheath liquid channels 135 open only to the top surface of main channel 131 (see
Except for the portion of the confluence part with main channel 131, second sheath liquid channel 135 is composed of third groove 116 formed in substrate 110 and a part of film 120 that closes the opening of third groove 116 (see
The confluence part of second sheath liquid channel 135 with main channel 131 is composed of a part of one surface of substrate 110 (the surface on which film 120 is joined), a part of the opening of first groove 112 (the groove that makes up main channel 131), and curved portion 121 of film 120 (see
Note that while the confluence part of two first sheath liquid channels 133 is located upstream of the confluence part of two second sheath liquid channels 135 in main channel 131 in the present embodiment, the positions of these confluence parts are not limited to this. For example, the confluence part of two second sheath liquid channels 135 may be disposed upstream of the confluence part of two first sheath liquid channels 133. In addition, the confluence part of two first sheath liquid channels 133 may face the confluence part of two second sheath liquid channels 135.
Liquid ejection part 136 is a bottomed recess for removing the sheath liquid and the main liquid flowing in from main channel 131 in the sheath flow state. In the present embodiment, liquid ejection part 136 is composed of fourth through hole 117 formed in substrate 110 and a part of film 120 that closes one opening of fourth through hole 117 (see
Next, with reference to
The main liquid is introduced to main liquid introduction part 130, and the sheath liquid is introduced to two first sheath liquid introduction parts 132 and two second sheath liquid introduction parts 134. In this state, liquid ejection part 136 is depressurized, or a pressure is exerted on main liquid introduction part 130, two first sheath liquid introduction parts 132 and two second sheath liquid introduction parts 134. In this manner, the main liquid in main liquid introduction part 130 flows in main channel 131 toward liquid ejection part 136. In addition, the sheath liquid in first sheath liquid introduction part 132 flows in first sheath liquid channel 133 toward main channel 131, and the sheath liquid in second sheath liquid introduction part 134 flows in second sheath liquid channel 135 toward main channel 131.
Observation or fluorescence observation of main liquid 140 may be performed in the state where the sheath flow is formed in main channel 131 as illustrated in
As described above, liquid handling device 100 according to the present embodiment can stably form a sheath flow in which the sheath liquid surrounds the main liquid with a configuration that can be manufactured with film 120 at low cost. In addition, the volume ratio of the main liquid and the sheath liquid can be readily controlled by adjusting the shape of curved portion 121.
In addition, in fluorescence observation of the main liquid in main channel 131, the influence of autofluorescence can be reduced since film 120 is used in place of a substrate, and the degree of freedom of the working distance can be increased since film 120 has a small thickness.
Liquid handling device 200 according to Embodiment 2 is different from liquid handling device 100 according to Embodiment 1 only in configuration of curved portion 221. The same components as those of liquid handling device 100 according to Embodiment 1 are denoted by the same reference numerals, and the descriptions thereof are omitted.
As illustrated in these drawings, liquid handling device 200 includes substrate 110 and film 220. In substrate 110, a groove configured to be a channel and a through hole configured to be an inlet or an outlet are formed. Film 220 is joined to one surface of substrate 110 to close the openings of the groove and the through hole formed in substrate 110. A part of the region of film 220 is formed as curved portion 221 curved in a direction away from substrate 110. The groove of substrate 110 closed with film 220 serves as a channel for carrying main liquid or sheath liquid. In addition, the inner space of curved portion 221 also serves as a channel for carrying sheath liquid.
In the present embodiment, liquid handling device 200 includes main liquid introduction part 130, main channel 131, two first sheath liquid introduction parts 132, two first sheath liquid channels 133, two second sheath liquid introduction parts 134, two second sheath liquid channels 235, and liquid ejection part 136.
The upstream ends of two second sheath liquid channels 135 are connected to respective second sheath liquid introduction parts 134 different from each other, and the downstream ends of two second sheath liquid channels 135 are connected to main channel 131. In the present embodiment, two second sheath liquid channels 135 open only to the top surface of main channel 131 (see
Except for the portion of the confluence part with main channel 131, second sheath liquid channel 135 is composed of third groove 116 formed in substrate 110 and a part of film 220 that closes the opening of third groove 116 (see
The confluence part of second sheath liquid channel 135 with main channel 131 is composed of a part of one surface of substrate 110 (the surface on which film 220 is joined), a part of the opening of first groove 112 (the groove that makes up main channel 131), and curved portion 221 of film 220 (
Liquid handling device 200 according to Embodiment 2 can be used through a procedure similar to that of liquid handling device 100 according to Embodiment 1.
Liquid handling device 200 according to Embodiment 2 has an effect similar to that of liquid handling device 100 according to Embodiment 1.
Liquid handling device 300 according to Embodiment 3 is different from liquid handling device 100 according to Embodiment 1 only in configurations of second sheath liquid introduction part 334 and second sheath liquid channel 335. The same components as those of liquid handling device 100 according to Embodiment 1 are denoted by the same reference numerals, and the descriptions thereof are omitted.
As illustrated in these drawings, liquid handling device 300 includes substrate 310 and film 120. In substrate 310, a groove configured to be a channel and a through hole configured to be an inlet or an outlet are formed. Film 120 is joined to one surface of substrate 310 to close the openings of the groove and the through hole formed in substrate 310. A part of the region of film 120 is formed as curved portion 121 curved in a direction away from substrate 110. The groove of substrate 310 closed with film 120 serves as a channel for carrying main liquid or sheath liquid. In addition, the inner space of curved portion 121 also serves as a channel for carrying sheath liquid.
In the present embodiment, liquid handling device 300 includes main liquid introduction part 130, main channel 131, two first sheath liquid introduction parts 132, two first sheath liquid channels 133, two second sheath liquid introduction parts 334, two second sheath liquid channels 335, and liquid ejection part 136.
Two second sheath liquid introduction parts 334 are bottomed recesses to which the sheath liquid is introduced. In the present embodiment, each of two second sheath liquid introduction parts 334 is composed of third through hole 115 formed in substrate 110, and a part of film 120 that closes one opening of third through hole 115 (see
Two second sheath liquid channels 335 are channels in which liquid can move. The upstream ends of two second sheath liquid channels 335 are connected to respective second sheath liquid introduction parts 334 different from each other, and the downstream ends of two second sheath liquid channels 335 are connected to main channel 131. In the present embodiment, two second sheath liquid channels 335 open only to the top surface of main channel 131 (see
In the present embodiment, second sheath liquid channel 335 is composed of a part of one surface of substrate 310 (the surface on which film 120 is joined), a part of the opening of third through hole 115 (the through hole that makes up second sheath liquid introduction part 334), a part of the opening of first groove 112 (the groove that makes up main channel 131), and curved portion 121 of film 120 (see
Liquid handling device 300 according to Embodiment 3 can be used through a procedure similar to that of liquid handling device 100 according to Embodiment 1.
Liquid handling device 300 according to Embodiment 3 has an effect similar to that of liquid handling device 100 according to Embodiment 1.
As illustrated in these drawings, liquid handling device 400 includes substrate 410, first film 420 and second film 423. In substrate 410, a slit configured to be a channel and a through hole configured to be an inlet or an outlet are formed. First film 420 is joined to one surface of substrate 410 to close the openings of the slit and the through hole formed in substrate 410. A part of the region of first film 420 is formed as first curved portion 421 curved in a direction away from substrate 410. Second film 423 is joined to the other surface of substrate 410 to close the openings of the slit and the through hole formed in substrate 410. A part of the region of second film 423 is formed as second curved portion 424 curved in a direction away from substrate 410. The slit of substrate 410 closed with first film 420 and second film 423 serves as a channel for carrying the main liquid. In addition, the inner space of first curved portion 421 and the inner space of second curved portion 424 serve as channels for carrying the sheath liquid.
The thickness of substrate 410 is not limited. For example, the thickness of substrate 410 is 1 mm to 10 mm In addition, the material of substrate 410 is not limited. For example, the material of substrate 110 may be appropriately selected from publicly known resins and glass. Examples of the material of substrate 410 include polyethylene terephthalate, polycarbonate, polymethylmethacrylate, polyvinyl chloride, polypropylene, polyether, polyethylene, polystyrene, cycloolefin resin, silicone resin and elastomer.
The thickness of each of first film 420 and second film 423 is not limited as long as a required strength can be ensured. For example, each of first film 420 and second film 423 has a thickness of 30 μm to 300 μm. In addition, the material of first film 420 and second film 423 is not limited as long as first curved portion 421 or second curved portion 424 can be molded. For example, the material of first film 420 and second film 423 may be appropriately selected from publicly known resins. Examples of the material of first film 420 and second film 423 include polyethylene terephthalate, polycarbonate, polymethylmethacrylate, polyvinyl chloride, polypropylene, polyether, polyethylene, polystyrene, cycloolefin resin, silicone resin and elastomer. For observation of the main liquid flowing inside the channel, it is preferable that the material of at least one of first film 420 and second film 423 be transparent. In addition, for fluorescence observation of the main liquid flowing inside the channel, it is preferable that the material of at least one of first film 420 and second film 423 be a material that transmits excitation light and fluorescence, with low autofluorescence. First film 420 and second film 423 are joined to substrate 410 by, for example, thermal welding, laser welding, an adhesive agent and the like.
In the present embodiment, liquid handling device 400 includes main liquid introduction part 430, main channel 431, two first sheath liquid introduction parts 432, two first sheath liquid channels 433, two second sheath liquid introduction parts 434, two second sheath liquid channels 435, and liquid ejection part 436.
Main liquid introduction part 430 is a bottomed recess to which the main liquid is introduced. In the present embodiment, main liquid introduction part 430 is composed of first through hole 411 formed in substrate 410, first through hole 425 formed in second film 423, and a part of first film 420 that closes one opening of first through hole 411 (see
Main channel 431 is a channel in which liquid can move. The upstream end of main channel 431 is connected to main liquid introduction part 430, and the downstream end of main channel 431 is connected to liquid ejection part 436. In addition, as described later, first sheath liquid channel 433 and second sheath liquid channel 435 are connected to main channel 431 at respective positions different from each other. In the present embodiment, the main liquid is introduced from main liquid introduction part 430 to main channel 431, and the sheath liquid is introduced from the confluence part with first sheath liquid channel 433 and the confluence part with second sheath liquid channel 435. Accordingly, the main liquid flows in main channel 431 in the region from main liquid introduction part 430 to the confluence part with first sheath liquid channel 433, and the main liquid and the sheath liquid flow in main channel 431 in the region from the confluence part with first sheath liquid channel 433 to liquid ejection part 436. As described later, in the region from the confluence part with second sheath liquid channel 435 to liquid ejection part 436, the main liquid and the sheath liquid flow through main channel 431 in a sheath flow state in which the sheath liquid surrounds the main liquid.
Main channel 431 is composed of slit (slender through hole) 412 formed in substrate 410, a part of first film 420 that closes one opening of slit 412, and a part of second film 423 that closes the other opening of slit 412 (see
Two first sheath liquid introduction parts 432 and two second sheath liquid introduction parts 434 are bottomed recesses to which the sheath liquid is introduced. In the present embodiment, each of two first sheath liquid introduction parts 432 is composed of second through hole 413 formed in substrate 410, second through hole 426 formed in second film 423, and a part of first film 420 that closes one opening of second through hole 413 (see
Two first sheath liquid channels 433 are channels in which liquid can move. The upstream ends of two first sheath liquid channels 433 are connected to respective first sheath liquid introduction parts 432 different from each other, and the downstream ends of two first sheath liquid channels 433 are connected to main channel 431. The sheath liquid introduced to first sheath liquid introduction part 432 flows through first sheath liquid channel 433. First sheath liquid channel 433 opens at least to the bottom surface of main channel 431. In the present embodiment, the “bottom surface of a channel” means the surface on first film 420 side (the surface formed with first film 420) of the inner surfaces of the channel. In the present embodiment, two first sheath liquid channels 433 open only to the bottom surface of main channel 431 (see
In the present embodiment, two first sheath liquid channels 433 are composed of a part of one surface of substrate 410 (the surface on which first film 420 is joined), a part of the opening of second through hole 413 (the through hole that makes up first sheath liquid introduction part 432), a part of the opening of slit 412 (the slit that makes up main channel 431), and first curved portion 421 of first film 420 (see
Two second sheath liquid channels 435 are channels in which liquid can move. The upstream ends of two second sheath liquid channels 435 are connected to respective second sheath liquid introduction parts 434 different from each other, and the downstream ends of two second sheath liquid channels 435 are connected to main channel 431. The sheath liquid introduced to second sheath liquid introduction part 434 flows through second sheath liquid channel 435. Second sheath liquid channel 435 opens at least to the top surface of main channel 431. In the present embodiment, the “top surface of a channel” means the surface on second film 423 side (the surface formed with second film 423) of the inner surfaces of the channel In the present embodiment, two second sheath liquid channels 435 open only to the top surface of main channel 431 (see
In the present embodiment, two second sheath liquid channels 435 are composed of a part of the other surface of substrate 410 (the surface on which second film 423 is joined), a part of the opening of third through hole 415 (the through hole that makes up second sheath liquid introduction part 434), a part of the opening of slit 412 (the slit that makes up main channel 431), and second curved portion 424 of second film 423 (see
Note that while the confluence part of two first sheath liquid channels 433 is located upstream of the confluence part of two second sheath liquid channels 435 in main channel 431 in the present embodiment, the positions of these confluence parts are not limited to this. For example, the confluence part of two second sheath liquid channels 435 may be disposed upstream of the confluence part of two first sheath liquid channels 433. In addition, the confluence part of two first sheath liquid channels 433 may face the confluence part of two second sheath liquid channels 435.
Liquid ejection part 436 is a bottomed recess for removing the main liquid and the sheath liquid flowing in from main channel 431 in the sheath flow state. In the present embodiment, liquid ejection part 436 is composed of fourth through hole 417 formed in substrate 410, fourth through hole 427 formed in second film 423, and a part of first film 420 that closes one opening of fourth through hole 417 (see
Next, with reference to
The main liquid is introduced to main liquid introduction part 430 and the sheath liquid is introduced to two first sheath liquid introduction parts 432 and two second sheath liquid introduction parts 434. In this state, liquid ejection part 436 is depressurized, or a pressure is exerted on main liquid introduction part 430, two first sheath liquid introduction parts 432 and two second sheath liquid introduction parts 434. In this manner, the main liquid in main liquid introduction part 430 flows toward liquid ejection part 436 in main channel 431. In addition, the sheath liquid in first sheath liquid introduction part 432 flows toward main channel 431 in first sheath liquid channel 433, and the sheath liquid in second sheath liquid introduction part 434 flows toward main channel 431 in second sheath liquid channel 435.
Observation or fluorescence observation of main liquid 140 may be performed in the state where the sheath flow is formed in main channel 431 as illustrated in
Liquid handling device 400 according to Embodiment 4 has an effect similar to that of liquid handling device 100 according to Embodiment 1.
The liquid handling device of the embodiments of the present invention is useful for various uses such as laboratory tests, food tests, and environment tests, for example.
Number | Date | Country | Kind |
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2020-055818 | Mar 2020 | JP | national |