Micro nozzle device

Abstract

To provide a micro nozzle device which discharges a fluid to a plurality of micro regions and is capable of controlling the fluid in various forms.SOLUTION: A micro nozzle device comprises: three or more discharging capillary tubes arranged so that their discharge openings are mutually adjacent; two or more suctioning capillary tubes arranged so that their suction openings are adjacent to the discharge openings; a discharge pump for discharging a fluid from the discharging capillary tubes; and a suction pump for suctioning a fluid from the suctioning capillary tubes. A respective opening diameter of the discharge openings and the suction openings is 1 mm or smaller.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a micronozzle device capable of discharging and sucking a fluid into a minute area. [Background technology]

[0002] In the past, it was common to irradiate a caged compound with laser light in order to locally stimulate living cells using drugs. A caged compound is a compound in which a photodegradable protective group (cage) is bound to a biologically active substance, and the protective group is degraded by irradiation with visible or infrared light, resulting in the expression of the biological activity. Examples of biologically active substances that can be bound to the cage include neurotransmitters, low-molecular-weight ligands, and second messengers. By changing the molecular structure of such caged compounds with laser light or degrading the molecules (uncaging), the resulting molecules can be used to locally stimulate living cells.

[0003] However, while the method of irradiating a caged compound with a laser beam can stimulate a size range of several tens of micrometers by limiting the area where the laser beam is focused, there are problems in that the types of molecules that can be applied are limited, and that the caged compound before the reaction and the molecules generated (dissociated) after the reaction can affect living cells. In addition, it is necessary to use a costly and complex device equipped with a laser light source and a photodetector.

[0004] On the other hand, it is possible to locally stimulate biological cells by flowing a drug through a microchannel, but this requires that the biological cells be grown on the microchannel, and it is difficult to stimulate a desired location within the cell by flowing a drug.

[0005] On the other hand, photolithography has been widely used for the microfabrication of silicon substrates, but microfabrication on uneven surfaces is difficult because it is difficult to maintain a constant distance from the photomask (reticle), making surface processing on the order of micrometers difficult. Furthermore, there are no methods for microfabrication or localized synthetic reactions without using photolithography.

[0006] In view of the above, there has been a demand for a discharge nozzle device that can discharge a fluid onto an arbitrary minute area and limit the range over which the fluid acts to that minute area. As an example of such a discharge nozzle device, Non-Patent Document 1 discloses a chemical pen with four micro nozzles arranged in a ring shape. It is said that such a chemical pen makes it possible to restrict a chemical solution or the like to act in a minute area. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Sifeng Mao, et al. Chemical Science, 9, 10, 2081 (2019). Summary of the Invention [Problem to be solved by the invention]

[0008] However, the chemical pen disclosed in Non-Patent Document 1 had four or fewer micronozzles, making it difficult to control the interface of two or more reaction fields (microareas) with high precision, and there was a demand for a nozzle device that could control fluid in a more diverse manner in a microarea.

[0009] The present invention has been made in consideration of the above circumstances, and has an object to provide a micronozzle device that is capable of ejecting fluid onto a plurality of minute regions and controlling the fluid in a variety of forms. [Means for solving the problem]

[0010] That is, in order to solve the above problems, the present invention proposes the following means. The micronozzle device of the present invention comprises three or more discharge capillaries, one end of which is a discharge opening, two or more suction capillaries, one end of which is a suction opening, a discharge pump that discharges fluid from the discharge capillaries, and a suction pump that sucks fluid from the suction capillaries, and is characterized in that the opening diameter of each of the discharge openings and the suction openings is 1 mm or less.

[0011] According to the present invention, by using three or more discharge capillaries each having a discharge opening at one end and two or more suction capillaries each having a suction opening at one end, fluid is discharged from three or more discharge openings and fluid is sucked from two or more suction openings, thereby making it possible to form linear or dot-like fluid regions in a minute region that was difficult to form with the conventional four or less discharge capillaries and suction capillaries. In addition, by providing two or more suction capillaries, it becomes possible to control the flow rate of the suction capillaries, and therefore it becomes possible to control the size of the fluid region at a desired position with higher accuracy, making it possible to form smaller linear or dot-like regions.

[0012] In addition, by individually setting the amount of fluid discharged from each discharge opening and the amount of fluid sucked in from each suction opening, it becomes possible to form multiple fluids into any desired shape in a small area.

[0013] In the present invention, the discharge openings may be arranged along a line, and the suction openings may be arranged on either side of the line.

[0014] In addition, in the present invention, the number of the discharge capillaries may be three, the number of the suction capillaries may be two, and the discharge opening of one of the three discharge capillaries may be arranged in a position surrounded by the discharge openings of the other two discharge capillaries and the suction openings of the two suction capillaries.

[0015] In the present invention, the discharge capillary and the suction capillary may be made of a material containing silicon.

[0016] In addition, in the present invention, the discharge capillary may be formed so that its inner diameter is larger than the opening diameter of the discharge opening upstream of the discharge opening, and the suction capillary may be formed so that its inner diameter is larger than the opening diameter of the suction opening upstream of the suction opening.

[0017] In addition, in the present invention, the fluid may be a drug solution for manipulating living cells. Effect of the Invention

[0018] According to the present invention, it is possible to provide a micronozzle device capable of ejecting a fluid onto a plurality of minute regions and controlling the fluid in various forms. [Brief description of the drawings]

[0019] [Figure 1] FIG. 1 is an enlarged perspective view of a main portion of a micronozzle device according to an embodiment of the present invention. [Diagram 2] FIG. 2 is a schematic diagram showing an arrangement of discharge openings and suction openings of a micronozzle device. [Diagram 3] 13 is a photograph showing the results of cell staining. [Figure 4] 13 is a graph showing the relationship between the formation width of fluorescein discharged from a central discharge opening and the flow rate ratio. [Diagram 5] 1 is a photograph showing the fluorescein formation shape (fluorescent area). [Figure 6] 1 is a photograph showing the fluorescein formation shape (fluorescent area). [Figure 7] This is a photograph showing the position where a fluorescent dye is linearly formed on a cell (the nozzle device and fluorescent dye are illustrated). [Figure 8] The photographs show the diffusion state of the fluorescent dye within the cells 5 minutes after the cells were stained. [Figure 9]The photographs show the diffusion state of the fluorescent dye within the cells 15 minutes after staining of the cells. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] Hereinafter, a micro nozzle device according to an embodiment of the present invention will be described with reference to the drawings. Each embodiment shown below is specifically described to better understand the gist of the invention, and does not limit the present invention unless otherwise specified. Furthermore, the drawings used in the following description may show essential parts enlarged for convenience in order to make the features of the present invention easier to understand, and the dimensional ratios of each component may not necessarily be the same as the actual ones.

[0021] Fig. 1 is an enlarged perspective view of a main portion of a micronozzle device according to one embodiment of the present invention, and Fig. 2 is a schematic diagram showing the arrangement of the discharge openings and suction openings of the micronozzle device. The micronozzle device 10 of this embodiment has first to third discharge capillaries 11, 12, 13, discharge pumps 21, 22, 23 which supply fluid to the first to third discharge capillaries 11, 12, 13, respectively, to discharge the fluid, first and second suction capillaries 15, 16, and suction pumps 25, 26 which suck fluid from the first and second suction capillaries, respectively.

[0022] The first to third discharge capillaries 11, 12, and 13 are, for example, hollow capillaries made of glass, with one end (opening end) serving as discharge openings 11a, 12a, and 13a, respectively. The first and second suction capillaries 15 and 16 are, for example, hollow capillaries made of glass, with one end (opening end) serving as suction openings 15a and 16a, respectively. The first to third discharge capillaries 11, 12, 13 and the first and second suction capillaries 15, 16 are bundled together near one end.

[0023] The first to third discharge capillaries 11, 12, 13 and the first and second suction capillaries 15, 16 may be made of a vitreous material containing silicon, such as fused silica, in addition to glass.

[0024] The discharge openings 11a, 12a, 13a and the suction openings 15a, 16a are open on the same plane. In this embodiment, the discharge openings 11a, 12a, 13a are arranged to be aligned on a line L. The suction openings 15a, 16a are formed at the position of the discharge opening 13a so as to sandwich the line L between them. That is, in the micronozzle device 10 of this embodiment, the discharge openings 11a, 12a and the suction openings 15a, 16a are formed to surround the discharge opening 13a with the discharge opening 13a at the center.

[0025] The discharge openings 11a, 12a, 13a of the first to third discharge capillaries 11, 12, 13 may have an opening diameter of 1 mm or less, for example, about 0.5 mm to 0.01 mm. The suction openings 15a, 16a of the first and second suction capillaries 15, 16 may have an opening diameter of 1 mm or less, for example, about 0.5 mm to 0.01 mm. The discharge openings 11a, 12a, 13a and the suction openings 15a, 16a may have different opening diameters from each other.

[0026] In this embodiment, the diameters of the discharge openings 11a, 12a and the suction openings 15a, 16a are 0.32 mm, and the diameter of the central discharge opening 13a is 0.05 mm. The outer diameters of the first and second discharge capillaries 11, 12 and the first and second suction capillaries 15, 16 are about 0.42 mm, and the outer diameter of the third discharge capillary 13 is about 0.15 mm.

[0027] The discharge pumps 21, 22, 23 and the suction pumps 25, 26 may be selected according to the type of fluid, and may be, for example, a syringe. Note that the discharge pumps 21, 22, 23 and the suction pumps 25, 26 may be, for example, a tube pump, other than a syringe.

[0028] The fluid discharged and sucked by the micronozzle device 10 of this embodiment may be any of liquid, sol, and gas. For example, when used for locally stimulating and observing living cells, a dye or an activator can be used as the fluid. When forming a nano-wiring pattern of any shape, a conductive paste such as gold or silver can be used as the fluid. When observing the action of a reactive gas locally on a sample, a reactive gas can be used as the fluid.

[0029] The operation of the micronozzle device 10 having the above configuration will now be described. As shown in FIG. 2, for example, when using a micronozzle device 10 to stimulate a biological cell M and observe the change thereof, a linear fluid region G is formed by aligning the discharge opening 13a of the third discharge capillary 13 with the position where the fluid region G is to be formed, and then discharging fluid from the discharge openings 11a, 12a, 13a of the first to third discharge capillaries 11, 12, 13, respectively, and sucking the discharged fluid from the suction openings 15a, 16a of the first and second suction capillaries 15, 16, respectively.

[0030] As a result, the fluid discharged from the central discharge opening 13a is sandwiched between the fluids discharged from the discharge openings 11a and 12a, and does not spread in the horizontal direction x in Fig. 2, and is pulled in the vertical direction y in Fig. 2 by the suction of the fluid by the suction openings 15a and 16a. This makes it possible to form a linear fluid region G that cuts across the biological cell M.

[0031] In this way, by using the first to third discharge capillaries 11, 12, 13 and the first and second suction capillaries 15, 16 to discharge fluid from three discharge openings 11a, 12a, 13a and suck in fluid from two suction openings 15a, 16a, it is possible to form a linear fluid region G in a very small area, which was difficult to form with four or fewer discharge and suction capillaries in the conventional method.

[0032] In this embodiment, the linear fluid region can be bent in the lateral direction x by making the discharge amounts of fluid from the discharge openings 11a and 12a different from each other. Also, by making the discharge amounts of fluid discharged from the discharge openings 11a, 12a, and 13a and the suction amounts of fluid sucked into the suction openings 15a and 16a different from each other, a plurality of fluids can be formed in a small region to have any shape.

[0033] Furthermore, by making the viscosities of the fluids discharged from the respective discharge openings 11a, 12a, and 13a different from one another, it is possible to form a plurality of fluids into a minute region so that each of the fluids has an arbitrary shape.

[0034] In this embodiment, an example is shown in which a micronozzle device is configured with a total of five capillaries, three discharge capillaries and two suction capillaries, but a micronozzle device can also be configured with four or more discharge capillaries or three or more suction capillaries. By providing four or more discharge capillaries or three or more suction capillaries, it becomes possible to lay out multiple fluids in various shapes in a small area.

[0035] Although the embodiments of the present invention have been described above, these embodiments are presented as examples and are not intended to limit the scope of the invention. These embodiments can be implemented in various other forms, and various omissions, substitutions, and modifications can be made without departing from the spirit of the invention. These embodiments and their modifications are included in the scope of the invention and its equivalents as described in the claims, as well as in the scope and spirit of the invention. EXAMPLES

[0036] Example 1 Cells were stained using the micronozzle device 10 of the above-described embodiment. Fluorescein solution (concentration: 10 μmol / L) was discharged from the discharge opening 13a located in the center at a flow rate of 5 μL / h, and phosphate buffer solution (pH 7.4) was discharged from the discharge openings 11a and 12a on both sides at a flow rate of 600 μL / h, respectively, and the discharged fluid was aspirated from the suction openings 15a and 16a on either side of the discharge opening 13a by changing the suction flow rate to 600, 1200, 1800, and 2100 (μL / h). Figure 3 shows the result of staining the cells. According to Figure 3, a linear flow layer of fluorescein solution was formed in the center.

[0037] 4 is a graph showing the relationship between the width of the fluorescein liquid discharged from discharge opening 13a and the flow rate ratio, where the flow rate ratio represents the ratio between the amount of fluid sucked in from suction opening 15a and the amount of fluid sucked in from suction opening 16a.

[0038] According to the results shown in FIG. 4, the width of the fluorescein liquid formed in the center decreases with an increase in the flow rate ratio. This confirmed that the fluid discharged from the discharge opening 13a can be formed into a line of any width by changing the ratio between the amount of fluid sucked in from the suction opening 15a and the amount of fluid sucked in from the suction opening 16a. This means that in addition to the conventional method of controlling the width of the fluid by changing the amount of fluid discharged from the discharge opening and the amount of fluid sucked in from the suction opening, there is a new method of controlling the width of the fluid. This confirmed that the width of the fluid can be controlled with higher precision.

[0039] Example 2 The spread of the fluorescent dye fluid discharged from the central discharge opening 13a was examined using the micronozzle device 10 of the above-mentioned embodiment. Fluorescein liquid (fluorescent dye: calcein-AM (concentration: 10 μmol / L)) was discharged from the central discharge opening 13a at a flow rate of 5 μL / h, and fluid was discharged from the discharge openings 11a and 12a on both sides at a flow rate of 600 μL / h, respectively, and the suction flow rate of one suction opening 15a sandwiching the discharge opening 13a was changed to 600 μL / h, and the suction flow rate of the other suction opening 16a was changed to 600 μL / h and 1200 μL / h, and the formed shape (fluorescent area) of the discharged fluorescein liquid was observed.

[0040] The shape of the fluorescein liquid (fluorescent area) formed when the suction flow rate of the suction opening 16a is set to 600 μL / h is shown in the photograph of Fig. 5. Also, the shape of the fluorescein liquid formed when the suction flow rate of the suction opening 16a is set to 1200 μL / h is shown in the photograph of Fig. 6.

[0041] According to the results shown in Figures 5 and 6, it was confirmed that the area and size of the fluorescent region formed can be arbitrarily controlled by changing the suction amounts of suction openings 15a and 16a.

[0042] Example 3 Using the micronozzle device 10 of the above-described embodiment, a linear fluorescent dye region was formed at the end of a living cell, and the penetration state of the fluorescent dye was observed. Observation object: PC-12 cells The central outlet 13a discharges a fluorescent dye solution (calcein-AM (concentration: 1 μmol / L)) at a flow rate of 5 μL / h. Phosphate buffer solution (pH 7.4) is discharged from the discharge openings 11a and 12a on both sides at a flow rate of 600 μL / h. -Suction of fluid from suction openings 15a and 16a at a flow rate of 600μL / h

[0043] FIG. 7 is a photograph showing the position of the fluorescent dye line on the cell. The diffusion state of the fluorescent dye within the cells 5 minutes and 15 minutes after staining is shown in FIG. 8 and FIG. 9, respectively.

[0044] 8 and 9, it was confirmed that the edge of the cell was appropriately stained and the fluorescent dye was diffused to the cell body by using the micronozzle device 10. The effectiveness of the micronozzle device of the present invention was confirmed. [Explanation of symbols]

[0045] 10. Micro nozzle device 11...First discharge capillary 12...Second discharge capillary 13…Third discharge capillary 11a, 12a, 13a...Discharge opening 15…First suction capillary 16…Second suction capillary 15a,16a...Suction opening 21, 22, 23...Discharge pump 25,26…Suction pump

Claims

1. The device includes three or more discharge capillaries, one end of which is a discharge opening, two or more suction capillaries, one end of which is a suction opening, a discharge pump that discharges a fluid from the discharge capillaries, and a suction pump that sucks a fluid from the suction capillaries, The micronozzle device is characterized in that the discharge opening and the suction opening each have a diameter of 1 mm or less.

2. 2. The micronozzle device according to claim 1, wherein the discharge openings are arranged along a line, and the suction openings are arranged on either side of the line.

3. The number of the discharge capillaries is three, and the number of the suction capillaries is two, 3. The micronozzle device according to claim 1, wherein the discharge opening of one of the three discharge capillaries is disposed at a position surrounded by the discharge openings of the other two discharge capillaries and the suction openings of the two suction capillaries.

4. 4. The micronozzle device according to claim 1, wherein the discharge capillary and the suction capillary are made of a material containing silicon.

5. 5. The micronozzle device according to claim 1, wherein the discharge capillary is formed so that an inner diameter thereof is larger than an opening diameter of the discharge opening on an upstream side of the discharge opening, and the suction capillary is formed so that an inner diameter thereof is larger than an opening diameter of the suction opening on an upstream side of the suction opening.

6. 6. The micronozzle device according to claim 1, wherein the fluid is a medicinal liquid for manipulating living cells.

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