This application is entitled to and claims the benefit of Japanese Patent Application No. 2020-165250, filed on Sep. 30, 2020, and No. 2020-218772, filed on Dec. 28, 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 fluid handling device.
In recent years, microwell plates, channel chips, and the like have been used for analysis of cells, proteins, nucleic acids, and the like. Advantageously, the microwell plates and channel chips require only a small amount of reagents and samples for the analysis, and are expected to be used in various applications such as clinical tests, food tests, and environmental tests.
For example, Patent Literature (hereinafter referred to as “PTL”) 1 discloses that a microwell plate (sample processing plate) is used to extract DNA using magnetic beads from a sample such as blood.
The present inventors have considered that a fluid handling device can be used in a versatile manner with various modifications for providing specific functions, while having a common structure.
An object of the present invention is to provide a fluid handling device having a rotary membrane valve and a rotary membrane pump, and further having a specific function.
A fluid handling device according to a first invention of the present application is a fluid handling device that includes a substrate and a film bonded to the substrate and that is configured to process a fluid, the fluid handling device including: a channel; a well connected to the channel; a rotary membrane valve disposed between the channel and the well; and a rotary membrane pump connected to the channel, in which the substrate or the film has a structure for pulverizing an introduced sample at a position corresponding to the rotary membrane pump.
A fluid handling device according to a second invention of the present application is a fluid handling device that includes a substrate and a film bonded to the substrate and that is configured to process a fluid, the fluid handling device including: a channel; a well connected to the channel; a rotary membrane valve disposed between the channel and the well; and a rotary membrane pump connected to the channel; and a filter disposed in the well or between the well and the rotary membrane valve for separating a blood cell component.
A fluid handling device according to a third invention of the present application is a fluid handling device that includes a substrate and a film bonded to the substrate and that is configured to process a fluid, the fluid handling device including: a channel; a well connected to the channel; a rotary membrane valve disposed between the channel and the well; a rotary membrane pump connected to the channel; a gel disposed in the channel; and a pair of electrodes disposed to be capable of applying a voltage to the gel.
A fluid handling device according to a fourth invention of the present application is a fluid handling device that includes a substrate and a film bonded to the substrate and that is configured to process a fluid, the fluid handling device including: a channel; a well connected to the channel; a rotary membrane valve disposed between the channel and the well; and a rotary membrane pump connected to the channel, in which the channel includes a fluorescence detection section that is a space for accommodating the fluid on which detection of fluorescence is performed, and the fluorescence detection section is deeper than a portion of the channel other than the fluorescence detection section.
A fluid handling device according to a fifth invention of the present application is a fluid handling device that includes a substrate and a film bonded to the substrate and that is configured to process a fluid, the fluid handling device including: a channel; a well connected to the channel; a rotary membrane valve disposed between the channel and the well; and a rotary membrane pump connected to the channel, in which the channel includes a branch portion and a liquid surface detection section that is disposed at a predetermined distance from the branch portion and at which arrival of a liquid surface is sensed, and a cross-sectional area of a portion of the channel between the branch portion and the liquid surface detection section is smaller than a cross-sectional area of a portion of the channel not situated between the branch portion and the liquid surface detection section.
A fluid handling device according to a sixth invention of the present application is a fluid handling device that includes a substrate and a film bonded to the substrate and that is configured to process a fluid, the fluid handling device including: a channel; a well connected to the channel; a rotary membrane valve disposed between the channel and the well; and a rotary membrane pump connected to the channel, in which the rotary membrane pump includes an agar medium disposed between the substrate and the film.
A fluid handling device according to a seventh invention of the present application is a fluid handling device that includes a substrate and a film bonded to the substrate and that is configured to process a fluid, the fluid handling device including: a channel; a well connected to the channel; a rotary membrane valve disposed between the channel and the well; and a rotary membrane pump connected to the channel, in which a surface of the substrate exposed in the channel or the well or a surface of the film exposed in the channel or the well includes an immobilization surface for immobilization of an antibody or an antigen, and the immobilization surface of the substrate or the film has a surface physical property different from that of a surface other than the immobilization surface.
According to the present invention, it is possible to provide various fluid handling devices having a common structure of a rotary membrane valve and a rotary membrane pump, and further having specific functions.
Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings.
[Configuration of Fluid Handling Device]
As illustrated in
Fluid handling device 200 includes substrate 210 and film 220 bonded to substrate 210. Further, in the present embodiments, fluid handling device 200 includes common channel 240, a plurality of wells 230 connected to common channel 240, a plurality of rotary membrane valves 232 disposed between common channel 240 and the plurality of wells 230, and rotary membrane pump 251 connected to common channel 240, and is capable of processing a fluid.
First rotary member 110 is rotated about first central axis CA1 by an external drive mechanism (not illustrated). Second rotary member 120 is rotated about second central axis CA2 by an external drive mechanism (not illustrated).
Fluid handling device 200 includes substrate 210 and film 220. The channel (common channel 240) is formed between substrate 210 and film 220. For example, a groove serving as the channel, through holes serving as the wells (inlets or outlets of the fluid), and the like are formed in substrate 210. Film 220 is bonded to one surface of substrate 210 to close the opening of the groove and one openings of the through holes. In addition, a plurality of diaphragm portions 250 are formed on a part of film 220. Film 220 may be a single layer film or a laminate of a plurality of films. For example, film 220 may be a laminate of a first film and a second film.
For example, two channels are formed by closing, with film 220, openings of two grooves formed in substrate 210. A portion of film 220 located on a partition wall (a portion of substrate 210 between the two grooves) formed between these two channels is caused to be deflected on the side opposite to substrate 210 to form diaphragm portion 250, whereby a valve (diaphragm valve or membrane valve) having diaphragm portion 250 between the two channels can be formed (see diaphragm portion 250 on the right side in
In addition, a portion of film 220 located in an arc-shaped region of substrate 210 in which no groove is formed is caused to be deflected on the side opposite to substrate 210 to serve as diaphragm portion 250, whereby rotary membrane pump 251 having diaphragm portion 250 can be formed (see diaphragm portion 250 on the left side in
Each of first rotary member 110 and second rotary member 120 has convex portion 111 for pressing diaphragm portion 250.
First rotary member 110 rotates about first central axis CA1, second rotary member 120 rotates about second central axis CA2. By rotation of each of first rotary member 110 and second rotary member 120, convex portion 111 comes into or goes out of contact with diaphragm portion 250. For example, the valve closes when convex portion 111 comes into contact with diaphragm portion 250 of the valve, and the valve opens when convex portion 111 is away from diaphragm portion 250 of the valve. Further, when convex portion 111 presses rotary membrane pump 251 in the direction from one end to the other end (e.g., clockwise), the fluid moves from one end to the other end, and when convex portion 111 presses rotary membrane pump 251 in the direction from the other end to one end (e.g., counterclockwise), the fluid moves from the other end to one end. This controls the movement of the fluid in fluid handling device 200. Note that, in the illustration of
Embodiments 1 to 7 will be described below as application examples of fluid handling device 200.
A description will be given of Embodiment 1 that is fluid handling device 200 having a function of pulverizing an introduced sample.
As described above, fluid handling device 200 according to Embodiment 1 includes common channel 240, a plurality of wells 230 connected to common channel 240, a plurality of rotary membrane valves 232 disposed between common channel 240 and a plurality of wells 230, and rotary membrane pump 251 connected to common channel 240.
In addition, in this fluid handling device 200, substrate 210 or film 220 has a structure for pulverizing an introduced sample at a position corresponding to rotary membrane pump 251. Examples of the structure for pulverizing the introduced sample include a roughened surface disposed on the surface of substrate 210 on the film 220 side at the position corresponding to rotary membrane pump 251, a plurality of protrusions disposed on the surface of substrate 210 on the film 220 side at the position corresponding to rotary membrane pump 251, a roughened surface disposed on the surface of film 220 on the substrate 210 side at the position corresponding to rotary membrane pump 251, a plurality of particles disposed between the first film and the second film constituting film 220 at the position corresponding to rotary membrane pump 251, and the like. One of these structures may be used alone or a plurality of the structures may be used in combination.
In the fluid handling device according to Embodiment 1, a sample (e.g., a formalin-fixed paraffin-embedded (FFPE) sample) is introduced into rotary membrane pump 251 from well 230 connected to rotary membrane pump 251. In this state, second rotary member 120 rotates to bring substrate 210 and film 220 into contact with each other at rotary membrane pump 251, whereby the sample between them is pulverized. For example, DNA is extracted from the pulverized sample.
Well 230 through which the sample is introduced and introduction channel 231 between well 230 and rotary membrane pump 251 are configured to be capable of receiving the sample. The cross-sectional size of introduction channel 231 is not particularly limited as long as introduction channel 231 is capable of receiving the sample, and the cross section has, for example, a rectangular shape with each side of a length equal to or greater than 1 mm.
According to fluid handling device 200 according to Embodiment 1, it is possible to pulverize the sample at rotary membrane pump 251
A description will be given of Embodiment 2 that is fluid handling device 200 having a function of removing blood cells from an introduced blood sample.
As described above, fluid handling device 200 according to Embodiment 2 includes common channel 240, a plurality of wells 230 connected to common channel 240, a plurality of rotary membrane valves 232 disposed between common channel 240 and a plurality of wells 230, and rotary membrane pump 251 connected to common channel 240.
Further, this fluid handling device 200 includes well 230 for introducing a blood sample or a filter for separating blood cells that is disposed between well 230 for introducing the blood sample and rotary membrane valve 232 corresponding to this well 230. The filter provided makes it possible for the fluid handling device according to Embodiment 2 to separate the blood cells and thus analyze plasma components. In particular, it is useful, for example, for analyzing cell-free DNA existing in plasma.
The configuration of the filter is not particularly limited as long as the filter is impermeable to blood cells and permeable only to plasma. For example, the filter may be a plurality of protrusions disposed on the surface of substrate 210 on the film 220 side in a channel between well 230 for introducing the blood sample and rotary membrane valve 232 corresponding to this well 230.
It is preferable that fluid handling device 200 according to Embodiment 2 include a liquid surface detection section that is disposed downstream of the filter and at which arrival of a liquid surface is sensed by an external device. By sensing the arrival of the liquid surface at the liquid surface detection section, it is possible to detect clogging of the filter obstructing the flow of the liquid.
According to fluid handling device 200 according to Embodiment 2, it is possible to separate blood cells by using the filter and to allow only plasma to flow through the channel.
A description will be given of Embodiment 3 that is fluid handling device 200 having a function of performing electrophoresis of a sample.
As described above, fluid handling device 200 according to Embodiment 3 includes common channel 240, a plurality of wells 230 connected to common channel 240, a plurality of rotary membrane valves 232 disposed between common channel 240 and a plurality of wells 230, and rotary membrane pump 251 connected to common channel 240.
Further, this fluid handling device 200 includes gel 260 and a pair of electrodes (anode 263 and cathode 264) for performing electrophoresis. In the present embodiment, as illustrated in
It is preferable that the depth and width of the channel filled with gel 260 be greater than the depth and width of a channel connected to the channel filled with gel 260. Thus, the volume of gel 260 can be increased as illustrated in
When electrophoresis is performed, for example, a solution containing a nucleic acid may be delivered under pressure so that the nucleic acid enters the inside of gel 260, and then a voltage may be applied.
As illustrated in
While the example has been described above in which a space between substrate 210 and film 220 is filled with gel 260, a space between the first film and the second film constituting film 220 may also be filled with gel 260. In this case, two through holes that function respectively as a gel inlet and a gel outlet are formed in one of the first film and the second film facing the channel. A sample flowing through the channel is applied a voltage upon arrival at the gel inlet, moves through the gel, and then arrives at the gel outlet. The arrival of the sample at gel 260 may be sensed, for example, with the configuration illustrated in
According to fluid handling device 200 according to Embodiment 3, it is possible to perform electrophoresis of a sample.
A description will be given of Embodiment 4 that is fluid handling device 200 capable of performing fluorescence detection on a sample.
As described above, fluid handling device 200 according to Embodiment 4 includes common channel 240, a plurality of wells 230 connected to common channel 240, a plurality of rotary membrane valves 232 disposed between common channel 240 and a plurality of wells 230, and rotary membrane pump 251 connected to common channel 240.
In addition, in this fluid handling device 200, the channel includes fluorescence detection sections 271 (271a to 271c) that are a space for accommodating a fluid on which detection of fluorescence is performed. In fluorescence detection sections 271 (271a to 271c), detection of fluorescence emitted from the fluid in the channel is performed using an external fluorescence detection device. In order to increase the fluorescence intensity to improve the detection sensitivity, the depth of fluorescence detection sections 271 (271a to 271c) is greater than the depth of a portion of the channel other than fluorescence detection sections 271 (271a to 271c). For example, the depth of fluorescence detection sections 271 (271a to 271c) can be increased by deepening grooves for forming fluorescence detection sections 271 (271a to 271c) that are formed in substrate 210 or by expanding film 220 forming fluorescence detection sections 271 (271a to 271c) to the side opposite to substrate 210. The widths of fluorescence detection sections 271 (271a to 271c) may be the same as or different from the width of the portion of the channel other than fluorescence detection sections 271 (271a to 271c). In the present embodiment, the widths of fluorescence detection sections 271 (271a to 271c) are the same as the width of the portion of the channel other than fluorescence detection sections 271 (271a to 271c).
As illustrated in
Further, wells 230a and 230b are connected to common channel 240 via common first channel 241. Similarly, wells 230c and 230d are connected to common channel 240 via common second channel 242. Wells 230e and 230f are connected to common channel 240 via common third channel 243. First channel 241 includes first fluorescence detection section 271a, second channel 242 includes second fluorescence detection section 271b, and third channel 243 includes third fluorescence detection section 271c. As illustrated in
Further, fluid handling device 200 also includes wells 230g to 230k. A reagent for reaction with a sample, for example, is introduced into wells 230g to 230k. Note that, as described above, rotary membrane valves 232 are disposed for wells 230g to 230k, respectively.
For example, the sample is introduced into well 230a, only valve 232a is opened, and the sample is caused to move to a position in front of common channel 240 by rotary membrane pump 251. Next, only valve 232g is opened, and the reagent is caused to move to common channel 240 by rotary membrane pump 251.
Then, only valve 232b is opened, and the reagent having moved to common channel 240 is caused to move toward well 230b by rotary membrane pump 251. It is thus possible to mix the sample and the reagent in well 230b. After the mixture and reaction, the reaction liquid may be moved to fluorescence detection section 271 (first fluorescence detection section 271a) for detection of fluorescence.
Like well 230a and well 230b described above, the pair of well 230c and well 230d and the pair of well 230e and well 230f are capable of handling respective different samples: a total of three samples can be handled.
According to fluid handling device 200 according to Embodiment 4, it is possible to perform fluorescence detection on a plurality of samples.
A description will be given of Embodiment 5 that is fluid handling device 200 capable of fractionating a liquid into a trace amount of fractions will be described.
As described above, fluid handling device 200 according to Embodiment 5 includes common channel 240, a plurality of wells 230 connected to common channel 240, a plurality of rotary membrane valves 232 disposed between common channel 240 and a plurality of wells 230, and rotary membrane pump 251 connected to common channel 240.
Further, in fluid handling device 200, as illustrated in
Measurement of a trace amount of liquid, e.g., at a level of pL and fractionation of the liquid with fluid handling device 200 according to Embodiment 5 will be described below with reference to
To begin with, a sample is introduced into well 230a, valve 232a is opened, and the sample is caused to move to liquid surface detection section 280 of common channel 240 by rotary membrane pump 251. The position of the liquid surface of the sample may be detected by the mechanism illustrated in
Here, since the space between the branch portion and liquid surface detection section 280 is narrow, an amount of sample on the order of picoliters, for example, can exist in this space.
Next, valve 232a is closed, and only valve 232b is opened, so that the sample existing in common channel 240 moves toward well 230b by rotary membrane pump 251. Accordingly, fractions 20 of the sample on the order of picoliters are obtained in the channel toward well 230b.
Repeating the above procedure results in multiple fractions 20 of the sample on the order of picoliters as illustrated in
According to fluid handling device 200 according to Embodiment 5, it is possible to fractionate a sample into a trace amount of fractions.
A description will be given of Embodiment 6 that is fluid handling device 200 capable of culturing bacteria and the like.
As described above, fluid handling device 200 according to Embodiment 6 includes common channel 240, a plurality of wells 230 connected to common channel 240, a plurality of rotary membrane valves 232 disposed between common channel 240 and a plurality of wells 230, and rotary membrane pump 251 connected to common channel 240.
Fluid handling device 200 according to Embodiment 6 includes an agar medium disposed between substrate 210 and film 220 at rotary membrane pump 251. The agar medium may be disposed in a groove formed in substrate 210 or may be disposed thinly on substrate 210. The type of agar medium is not particularly limited and may be appropriately selected depending on a cultured target.
According to fluid handling device 200 of Embodiment 6, it is possible to culture bacteria by introducing a bacterial suspension from well 230 connected to rotary membrane pump 251, and applying the introduced bacterial suspension onto the agar medium using second rotary member 120.
A description will be given of Embodiment 7 that is fluid handling device 200 capable of immobilization of an antibody or the like.
As described above, fluid handling device 200 according to Embodiment 7 includes common channel 240, a plurality of wells 230 connected to common channel 240, a plurality of rotary membrane valves 232 disposed between common channel 240 and a plurality of wells 230, and rotary membrane pump 251 connected to common channel 240.
In fluid handling device 200 according to Embodiment 7, a surface of substrate 210 exposed in the channel or the wells or a surface of film 220 exposed in the channel or the wells has an immobilization surface for immobilization of an antibody or an antigen. When substrate 210 is provided with the immobilization surface, the surface physical properties of the immobilization surface of substrate 210 are different from the surface physical properties of a surface of substrate 210 other than the immobilization surface. In addition, when film 220 is provided with the immobilization surface, the surface physical properties of the immobilization surface of film 220 are different from the surface physical properties of a surface of film 220 other than the immobilization surface. For example, the immobilization surface has surface physical properties allowing easier fixation of an antibody or an antigen, and the surface other than the immobilization surface has surface physical properties allowing a liquid to flow easily.
There is no particular limitation on a method of making the surface physical properties of the immobilization surface different from those of the other surface. For example, substrate 210 or film 220 may be partially masked and subjected to various treatments. In addition, the first film and the second film having surface physical properties different between the first and the second films may be laminated, and through holes may be formed in one of the films on the substrate side, so that the film to be exposed in the channel or the wells may be different for each place.
According to fluid handling device 200 according to Embodiment 7, it is possible to immobilize an antibody or an antigen in fluid handling device 200, to perform detection using the antibody or the antigen.
As described above, according to fluid handling devices 200 according to the embodiments of the present invention, it is possible to perform various processing. Fluid handling devices 200 according to the embodiments of the present invention can also be used for liquid processing in sample preparation for next generation sequencing (NGS) and for delivery of a liquid staining reagent for a culture medium and visualization in a cell assay.
The fluid handling devices of the present invention are useful in various applications such as, for example, clinical tests, food tests, and environmental tests.
Number | Date | Country | Kind |
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2020-165250 | Sep 2020 | JP | national |
2020-218772 | Dec 2020 | JP | national |