JP2011069831A - Optical detection device, analyzing apparatus, and droplet mixing method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To solve the problems wherein, a droplet operation method using a channel or a plane has a risk of contamination, and high cost, and further, when many droplets are to be conveyed, a device structure becomes complicated, and preparation or operation becomes difficult, and cost is heightened. <P>SOLUTION: A circular or spiral droplet holding part made of a wire rod is prepared, and droplets are held there in the suspended or included state. A means for moving the droplet holding part is added, to thereby convey the droplets. Two droplet holding parts are brought into contact each other to thereby mix the droplets, and the droplets are dropped by changing the shape of the wire rod of the droplet holding part from the outside. An optical path for allowing passage of the droplets therethrough is set, and optical measurement is performed. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention belongs to a method for conveying a minute amount of droplets. The present invention also belongs to the fields of generation and dispensing of microdroplets, chemical reactions using microdroplets, and substance generation using microdroplets. The present invention also belongs to the fields of chemical analysis and biochemical analysis, automatic blood analysis, and immunodiagnosis.

  Here, the prior art relating to the transport of a minute droplet related to the present invention will be described. The microdroplet as used in the present specification does not indicate the shape of the liquid, but is described below as an expression indicating a state in which the liquid is clearly separated from other liquids.

  There are various techniques for transporting microdroplets. For example, using a fine channel surrounded by four sides like a capillary or microfluidic MEMS (Mechanical and Electrical Micro System), the target droplet is sandwiched between gases and transported by power such as pressure difference. There is something to do. As a similar example, there is an example in which a liquid that does not mix with a target droplet instead of gas is sandwiched on both sides and conveyed in the same manner.

  Means for transporting microdroplets on a plane include EWOD (electrowetting on die electrode) and dielectrophoresis. In either case, the droplet is placed on a plane or sandwiched between two planes, and a controlled voltage is applied to an electrode built in or on the plane. A remote force is applied to the droplet to move the droplet.

  One that transports droplets in the air is one that uses a piezoelectric element to create and fly droplets. An example of applying this technology is a flow cytometer that fractions cells and the like. In this case, a small droplet generated by a piezoelectric element is loaded with electric charges according to the optical characteristics of the cells enclosed inside, and the flight curve of the droplet is generated by the installed magnetic field. Twist and bend to collect only the desired fraction. In addition, there is an example in which liquid is added and mixed by causing droplets to fly from two directions so as to cross.

  In addition to these methods, there is a method for transporting droplets by placing a small amount of droplets on another member. For example, there is a method in which a liquid is scooped with a ring of a transfer member having a surface with poor wettability, and a sphere-like liquid due to surface tension is transferred onto the ring of the transfer member (Patent Document 1). In addition, as part of the device configuration for depositing fluid dots on the surface, a liquid is held and moved like a film in a ring-shaped object, and a pin is pierced there to deposit the liquid attached to the tip of the pin There is a liquid handling method of performing (Patent Document 2).

JP 11-271185 A Special Table 2002-509274

  Problems to be solved by the present invention include the following. First, among the above-described conventional techniques, the method using a flow path or a flat surface has a problem that the liquid is contaminated by contact with the wall and the surface properties are changed. There is a problem of reducing the accuracy of the behavior and size control of the transported droplet, the reproducibility of the movement behavior, the efficiency of the final chemical reaction and the accuracy of the analysis, respectively. There is a problem that there is a possibility of contamination. This contamination can also cause problems such as chemical reactions and reduced accuracy of analysis and inspection. There is a problem that the flow path or flat surface used for liquid conveyance is expensive and expensive. In particular, the formation of fine channels and surfaces with electrodes is expensive. Therefore, it is difficult to take a method of disposable for solving the problems related to contamination such as disposable. In order to solve this problem, cleaning and reuse can be considered, but there is a limit to the recovery of the surface by normal cleaning. In addition, in the method of transporting droplets using a planar electrode, it is necessary to change the electrode arrangement when the size of the droplet changes, and conversely, the same electrode arrangement cannot handle droplets of greatly different sizes. Further, although the surface physical properties are greatly affected, it is difficult to achieve the reproducibility of the surface physical properties, and there is a problem in the accuracy of the droplet size. Liquid types that cannot be transferred by electrical means or solutions with severe surface contamination are difficult to handle, and liquid types also have problems.

  The method of transporting droplets in the air has a problem that there is an upper limit on the size of droplets that can be handled. When the droplet size exceeds a certain level, the force exerted by gravity increases and the falling speed increases. In addition, since there are no walls in the air, the problem of contamination is reduced, but the residence time cannot be taken, so it is difficult to use it for analysis / inspection means involving chemical reactions that require residence time.

  For any method that transports channels, planes, or air, a large number of droplets to be transported requires a complex device structure, which results in difficult device creation and operation. There is a problem that the cost becomes high. Another problem is that the configuration of the device must be changed each time the number of droplets to be transported changes.

  With regard to the droplet transport method taken up in the conventional technology other than the above, there is a problem in the size of the droplet that can be handled. In the “droplet transport method” described in (Patent Document 1), for example, since the radius of the ring is 0.8 mm, the radius of the droplet is calculated using the calculation formula in the text, and the volume is calculated therefrom. When calculated, it is considered that droplet transport of about 19 microliters or more cannot be performed. In addition, since the droplet holding part described in (Patent Document 2) is a liquid film, the surface diameter is 0.375 microns or less, and the pins are obviously smaller than that, for example, a liquid volume of 1 microliter or more is conveyed. This is a configuration that cannot be done. As with the size limitation, one of the problems is that droplets of different sizes cannot be handled without changing the configuration. In these configurations, it is difficult to perform operations such as droplet merging, droplet mixing, and droplet dropping.

  In order to solve the above problems, the present invention takes the following means. In the simplest configuration, an annular droplet holder made of wire is prepared, and droplets are held here. Unlike the “droplet conveying method” described in (Patent Document 1), it is not necessary to use a material having particularly poor wettability. For example, it is easy and inexpensive if a metal wire that is generally hydrophilic is used. It is. The liquid droplet is held by the liquid holding unit in a form of hanging from an annular wire. The retained droplets are stable, and can easily cope with, for example, an increase or decrease in the liquid amount by changing the droplet shape due to the effect of surface tension. By adding a means for moving the droplet holding portion to the annular droplet holding portion, the droplet conveyance is realized. For example, if an upward wire is attached to the annular droplet holder, and the handle that connects the upward wire to the stage is connected, the droplet can be transported by moving the stage. Can do.

  In the droplet transport method of the present invention, there is no major limitation on the material of the droplet holder, and the configuration is simple, so it is inexpensive and can be replaced. Further, unlike the method using a flow path or a flat surface, it is also advantageous that cleaning is easy. This is because when a flow path or a flat surface is used, the entire movement path is subject to contamination, whereas in the present invention, the surface area with which the liquid comes into contact is limited to the liquid holding portion, so it is overwhelmingly small. Whether it is replacement or cleaning, it becomes easier to deal with contamination issues. When using flow paths or flat surfaces, it is common for different solution types to pass through the same flow path or flat surface. However, in the present invention, appropriate wire materials and surface properties are selected for different solution types. Therefore, the accuracy of the solution size and the reproducibility of conveyance can be improved. Since it can be stably held and transported for a sufficiently long time, the problem that the residence time, which has been a problem particularly in the case of air transportation, is limited to be short is solved.

  Even if the number of droplets to be transported increases, it can be handled with a simple method of increasing the number of droplet holders, and the structure is simple, so the creation and operation is easy and the cost is high. There is no. It is only necessary to change the number of droplet holders used to change the number of droplets to be conveyed. Further, by adopting this configuration, it is possible to easily operate a droplet of about 20 microliters or more. It is also possible to manipulate minute droplets of 1 nanoliter.

  Regarding the mixing, the shape of the droplets can be changed by moving the droplet holding part up and down, left and right, and liquid flow can be caused inside the liquid, so that internal stirring can be easily performed. In addition, another droplet can be inserted into the liquid droplet held in the liquid holding portion, stirred and mixed, and a method of applying vibration with a vibrator is also effective.

  With regard to the merging of the liquids, the liquid droplets held in the liquid holding unit can be easily brought into contact with each other. The mixing is also automatically realized by the liquid flow during the merging. When the droplets are large and small, the small droplet has a higher internal pressure than the Laplace's law, so that the merging mode in which the small droplet is absorbed by the large droplet can be realized with good reproducibility. However, it is also possible to easily transfer a large droplet to a small droplet including a case where the droplets are merged in the vertical direction and a droplet holding portion described below is not circular.

  It is also possible to change the shape of the wire rod in the above-described droplet holding portion from the outside to reduce the droplet holding force and remove the droplet from the liquid holding portion, that is, drop it to a predetermined position. Of course, it is also possible to move the droplet by bringing it into contact with the location where the droplet itself is moved.

  Another simple droplet holder has a spiral shape. The liquid is held at a portion where the pitch of the helix is dense, and the liquid is not held when the pitch is wide to some extent. The advantage of this configuration is that it is easy to scoop out the liquid, which is difficult in the case of a simple ring. It is also possible to cope with a large amount of liquid. A predetermined liquid amount can be scooped up by attaching the spiral droplet holding portion to the solution reservoir and pulling it up again. Part or all of the liquid held in the spiral liquid holding part can be dropped at a predetermined position by widening the pitch of the spiral. It is conceivable to pull the tip of the spiral wire with another member, or to change the temperature of the spiral wire so as to eliminate the spiral if it is made of a shape memory alloy.

  Here, only the conveyance of the liquid droplets by the annular or spiral wire material has been described, but the shape of the liquid droplet holding part is not limited to this. The shape of the droplet holder can be appropriately changed according to the size and type of the droplet or the purpose of the droplet reaction. Compared to the case where a droplet is held and dropped using a pipette or the like, there is an advantage that an extra part such as a piston is not required, and for example, there is an advantage that a large number of droplet operations can be easily performed.

  With the above configuration, attachment, merging, mixing, stirring, holding for a long time, and easy scale-up are possible, so that it can be applied to generation of a minute droplet or a chemical reaction using the minute droplet. In addition, it can be applied to the fields of chemical analysis using droplets, biochemical analysis, and automatic blood analysis. For example, this method can be used for the spectroscopic cell portion of absorption measurement.

  According to the present invention, it is possible to convey droplets easily and inexpensively. Further, stirring, merging, mixing, and dropping of the droplets can be easily performed. Thereby, an inexpensive and simple configuration for handling droplets can be realized in the fields of chemical analysis, biochemical analysis, and automatic blood analysis. Even in applications where there is a contamination problem, it is possible to avoid the contamination problem by disposing the part in contact with the droplet. When applied to inspection, etc., the cost of inspection can be reduced. Furthermore, it is possible to easily cope with changes in the droplet size and the number of droplets, and a large number of uniform minute droplets can be created. When used in a spectroscopic cell, it is possible to perform measurement with a long optical path length with a small amount of liquid, and the sample can be recovered after the measurement.

The schematic diagram of the conveyance method of the droplet of one Example of this invention. The schematic diagram of the stirring of the droplet of one Example of this invention. The schematic diagram of the stirring of the droplet of one Example of this invention. The schematic diagram of the method of mounting the droplet of one Example of this invention on a droplet holding part. The schematic diagram of mixing of the droplet of one Example of this invention. The schematic diagram of the conveyance method of the droplet of one Example of this invention. FIG. 3 is a series of schematic views of holding, transporting, and dropping droplets according to an embodiment of the present invention. The schematic diagram of the dripping method of the droplet of one Example of this invention. The schematic diagram of the optical measuring device of one Example of this invention. The schematic diagram of the droplet collection | recovery method of one Example of this invention. The schematic diagram of the mass production method of the droplet of one Example of this invention. The schematic diagram of the droplet operation apparatus of one Example of this invention. The schematic diagram of the biochemical analyzer of the blood of one Example of this invention.

  Embodiments of the present invention will be described below with reference to the drawings. FIG. 1 is a schematic view of an embodiment of a droplet conveying means. An annular droplet holding portion 101 is provided with a moving handle (moving means) 103, which is composed of a wire. A droplet 102 is held in the annular droplet holding portion 101. The droplet 102 is inscribed macroscopically with respect to the annular droplet holding portion 101, and resists gravity by the surface tension of the liquid and holds the shape. The droplets are substantially spherical due to the surface tension, and are held in a hanging shape with respect to the annular droplet holding portion. For example, if a stainless steel wire with a diameter of 100 microns is used to create an annular holder having a diameter (inscribed diameter) of about 2.7 mm and hold water, a droplet of about 50 microliters of water can be held. If a droplet holder with a diameter of 3.2 mm is made using a 0.5 mm diameter solder wire, about 75 microliters of droplet can be held. By controlling the diameter of the annular portion, the diameter of the wire, and the surface properties in this way, the amount of water retained can be changed, and droplets of about 10 nanoliters to several hundred microliters can be retained. .

The amount of liquid retained can be estimated as follows. The droplet is held in contact with the annular droplet holder. Increasing the amount of the liquid droplets 102 held without changing the shape and size of the liquid holding portion 101 keeps a spherical shape under the annular droplet holding portion, and finally reaches the bottom of the holding portion only. It will be a little constricted, and it will fall further. Since the handle 103 in this embodiment is located above the annular droplet holding portion 101, it does not hinder the formation of droplets. Stable holding is possible only when the surface tension acting on the circumference of the diameter (inscribed diameter) inscribed in the droplet holding portion can hold the weight of the entire droplet. When the limit droplet is approximated by a sphere with a volume V and a diameter D, the inscribed diameter is d, the surface tension is γ, the density of the liquid is ρ, and the gravitational acceleration is g, the balance equation is 2πd × γ = V × ρ × g = (π / 6) × D ³ × ρ × g. Substituting 2.7 mm and 3.2 mm as the inscribed diameter d of the previous experiment and substituting the physical properties of water together, the volume V is estimated to be 63 and 75 microliters, respectively. In the case of the former with a thin wire diameter, the value is slightly smaller than the experimental value due to the stability problem, but the latter with the thick wire is exactly as calculated. The place where D and d are the same is the limit of this holding method, which is d = D = 6.7 mm. It is sufficiently possible to control the inner diameter of the annular droplet holder within the range of about 10 microns to this 6.7 mm, and the amount of liquid held at this time is calculated from 23 nanoliters to 156 microliters. This calculates the last holding amount that cannot be held, so in practice, even a droplet holding unit having a diameter of this size can hold a smaller droplet amount. In terms of the upper limit, the holding amount can be increased by making the ellipse out of the depth. In other words, this method can hold and transport droplets in the range of 10 nanoliters to sub-milliliters. For example, in the case of an annular holding portion having a diameter of 1 mm, it is possible to hold and transport a droplet of about 20 microliters.

  By moving the handle 103 attached to the droplet holder 101 in a three-dimensional manner, the droplets 102 held by the liquid holder 101 can be freely conveyed. There are various ways to move in three dimensions. A method of operating by gripping with a mechanical arm, a method of using an xyz stage, and the like are conceivable. By controlling the speed at which the droplets are transported so that the droplets do not fall due to movement, stable droplet transportation can be performed three-dimensionally. In the schematic diagram, the liquid holding unit is in a horizontal direction, but is not necessarily horizontal, and even if it is tilted, droplets can be held and transported.

  FIG. 2 is a schematic view of the stirring of the droplet. By grasping the handle 103 described with reference to FIG. 1 and moving it up and down, the states of FIGS. 2A and 2B are repeated, and the flow of liquid inside the droplet occurs, and the inside of the droplet can be stirred. The holding force of the droplet 101 by the annular droplet holding portion 102 is large, and the droplet 102 does not fall off even if it is moved lightly up and down. Agitation by this method is remarkably effective when the droplets are larger than a certain size, for example, when the size is 10 microliters or more. On the other hand, if it is smaller than this, the effect of molecular diffusion is sufficient, and stirring itself is often unnecessary. Although only the vertical movement is shown here, the same stirring effect can be obtained in the horizontal direction and the rotational movement.

  FIG. 3 is a schematic view in which droplets are stirred by different methods. FIG. 3A is a schematic view when a stirring rod is used. The tip of the stirring rod 104 that can penetrate the annular droplet holding portion 101 is inserted into the droplet 102, and mechanically mixed using the stirring motor 105 connected to the stirring rod 104 to perform stirring. FIG. 3B is a schematic diagram when using vibration. By applying vibration to the handle 103 by the eccentric motor 106, this vibration is transmitted to the droplet 102 via the liquid holding unit 101 and stirred. In this example, a motor is used as the stirring means and the vibration applying means, but the invention is not necessarily limited thereto.

  FIG. 4 is a schematic diagram of one method of placing a droplet on the droplet holding unit. First, as shown in FIG. 4A, the droplet holder 101 is attached to the container 110 containing the liquid to be held via the handle 103. Next, as shown in FIG. 4B, the droplet holding unit 101 is pulled up through the handle 103. At this time, a spherical droplet is not generated in the droplet holding unit 101, but a liquid film 111 is generated. Finally, as shown in FIG. 4 (d), when the solution is added to the liquid film 111 by the liquid supply means 113 (for example, a pipette), the droplet 112 has a center of gravity below the liquid holding portion 101, and is mainly lowered. It is held in a form that is convex to the side. The upper side of the droplet is also convex. The method of adding the solution from the liquid supply means 113 to the liquid film 111 does not pay particular attention and operates stably. It is also possible to adopt a form in which droplets are dropped from a small height, or a form in which the liquid is pushed out after the tip of the liquid supply means 113 is brought into contact with the liquid film 111. The method for placing the droplet on the droplet holder is not limited to this, and for example, the droplet can be directly placed on the droplet holder from the liquid supply means. This is possible when the size of the droplet to be placed is equal to or larger than that of the annular droplet holder. In some cases, a droplet may be generated by adding a liquid from the liquid supply means using the droplet as a core on the joint between the handle 103 and the droplet holding unit 101. The method of placing these droplets on the droplet holder is applicable, for example, when applying liquid to the tip of a pin that is part of the device configuration for depositing fluid dots on the surface described in Patent Document 2. This is one of the advantages of adopting the shape of the present invention. Further, when it is desired to discard the retained droplets, a gas such as air can be used to blow and blow off a gas such as air on the droplets. As a result, the droplet holder can be used multiple times.

  FIG. 5 is a schematic diagram showing mixing of droplets. In this embodiment, an example in which two large and small droplets are mixed by using a plurality of droplet transporting units having different diameters of the annular droplet holding portion is shown. FIG. 5A shows the state of two droplets before mixing. The large droplet 202 on the left side is held by the annular droplet holding unit 201 and can be conveyed via the handle 203. Similarly, the small droplet 212 on the right side is held by the annular droplet holding unit 211 and can be conveyed via the handle 213. Here, the annular droplet holder 201 has a larger diameter than the annular droplet holder 211. FIG. 5B shows a state in which these two droplets are brought close to each other and the droplets adhere to each other. The mixed droplet 220 is held in such a manner as to hang from both of the two annular holding portions 201 and 211. FIG. 5C is a diagram in which the mixed droplets 221 are transferred to the annular droplet holder 201 on one side by pulling the handles 203 and 213 away from the state of FIG. 5B. Little liquid remains in the small annular droplet holder 211, and only remains as a liquid film 222. The liquid film 222 does not always remain, and the liquid droplet 221 can be completely moved without generating the liquid film 222 depending on the angle of separation or the relative speed. Also, there is a method of completely transferring the droplet 221 by piercing a pin separately provided on the liquid film 222 at the time of separation to rupture the film. In this way, two droplets can be mixed and transferred to the droplet holding unit on one side. Due to the effect of Laplace pressure, a droplet with a small curvature has a higher internal pressure. Therefore, when droplets are merged from the side as in this embodiment, a small droplet is sucked into a large droplet. The droplets merge in a manner. However, it is also possible to easily transfer a large droplet to a small droplet including a case where the droplets are merged in the vertical direction and a droplet holding portion described below is not circular.

FIG. 6 is a schematic diagram showing a second embodiment of the droplet conveying means. Unlike the previous embodiments, the liquid holding portion has a spiral structure instead of an annular shape. A spiral droplet holder 301 is provided with a moving handle 303, which is made of a wire. A droplet 302 is held in the spiral droplet holding portion 301. The droplet 302 is inscribed macroscopically with respect to the annular droplet holding portion 301, and resists gravity by the surface tension of the liquid and holds the shape. The liquid droplet has a substantially spiral shape, and macroscopically touches the holding portion. Unlike the case where a droplet is suspended from an annular liquid holding unit, when a droplet is held over the entire length of the spiral droplet holding unit 301, a substantially constant amount of liquid is held. The liquid volume V of the droplet substantially matches the inner volume of the spiral droplet holder. The number of turns of the spiral n, d the diameter, the spacing of the wire p, that when V can substantially expressed as ^{V = πd 2/4 × p} × n. By changing these variables, it is possible to design a droplet holder that holds droplets having an arbitrary volume. It is possible to hold an amount of liquid equal to or greater than that of the annular droplet holder described in the embodiments so far. The protrusion 304 positioned below the spiral droplet holder 301 is not an essential component for transporting the droplet, but is a configuration effective when the droplet is released, which will be described later. The holding handle 303 for movement can carry the droplet 302 held by the droplet holding unit 301 freely by holding it and moving it three-dimensionally. This is the same as that described in Example 1. In terms of stability during transportation, this spiral holding section is higher, and more stable droplet transportation can be performed.

  FIG. 7 is a schematic diagram showing a series of states of holding, transporting, and dropping droplets in the second embodiment of the present invention. FIG. 7A shows a state in which no droplet is held and a container 310 containing a solution to be transported from now on. FIG. 7B shows a state in which the droplet holding unit 301 is attached to the solution in the container 310. When the droplet holder 301 is attached to the container 310 using the carrying handle 303, the solution enters the droplet holder 301. In this schematic diagram, the entire spiral droplet holder 301 is submerged in the solution, but the desired liquid can be obtained by applying only a part of the spiral droplet holder 301 to the solution by controlling the immersion height. It is also possible to hold a droplet having a quantity in the droplet holder 301. FIG. 7 (c) shows a state where this is pulled up. By pulling up, the droplet 302 is held inside the spiral droplet holding unit 301 and is not held in other portions. FIG. 7 (d) shows how the droplets are conveyed. The droplet 302 can be transported to a desired place by being moved through the handle 303. FIG. 7E is a schematic diagram in which the droplet 302 is moved to the well 311 of the microplate which is the movement destination. In this case, the projection 312 is a schematic diagram that has a structure protruding from the bottom of the well 311 of the microplate. Thereafter, the droplet holder 301 is lowered to drop the droplet 302 into the well 311 of the microplate. FIG. 7 (f) is a schematic diagram in which the protrusion 304 located below the droplet 302 is hooked on the protrusion 312 in the well 311 of the microplate. FIG. 7 (g) is a schematic diagram in which the handle 303 is pulled up while the protrusions 304 and 312 are hooked to drop the droplet 302 onto the well 311 of the microplate. When the spiral droplet holder 301 is stretched, the pitch of the spiral is expanded, and the droplet 302 held by the force of the surface tension of the liquid falls down by gravity and moves into the well 311 of the microplate. Thus, the liquid 313 in the microplate was obtained. FIG. 7 (h) shows a state in which the protrusions 304 and 312 are removed and the spiral droplet holder 301 is restored. By following the steps from FIGS. 7A to 7H, the droplets can be taken out from the container and moved to a predetermined location. Here, the hooking by the protrusion is used to stretch the spiral droplet holder, but the present invention is not limited to this method. Drops can be dropped by simple means such as pinching the tip with a tweezers and pulling, and a method using a shape memory alloy as in the following embodiment is also effective.

  FIG. 8 is a schematic diagram showing a droplet dropping method according to a third embodiment of the present invention. FIG. 8A shows a schematic diagram immediately before dropping. A droplet 402 is held in a spiral droplet holder 401 made of a shape memory alloy. The introduction of the liquid droplets into the liquid droplet holding section may be performed in the same manner as in the second embodiment, or a method of placing a liquid with the liquid supply means as in the first embodiment may be employed. A heater 404 connected to the controller 405 is attached to the handle 403 and controls the temperature of the droplet holder 402. When the controller 405 is operated so as to turn on the heater 404 in the state of FIG. 8A, the spiral droplet holder 402 extends to become a linear wire 407 as shown in FIG. 8B. Since the droplet 402 is not held by the linear wire 407, the liquid 408 is dropped on the bottom of the test tube 406. For this purpose, a shape memory alloy having a linear memory may be wound in a spiral shape, and in order to reduce problems such as evaporation and denaturation due to heating of the solution, for example, a memory temperature of about 30 to 40 degrees is used. Use shape memory alloys. In the schematic diagram of this embodiment, the spiral is completely extended and linear, but this is not always necessary. In order to reduce the possibility of droplets 402 splashing and contaminating other places when changing form from spiral to linear, here we put it inside a tall test tube 406 and then heat it However, this problem can be reduced by paying attention to the direction of the wire when winding in a spiral. Another effective method is to insert another wire into the spiral droplet holder 401 before dropping the droplet, and then drop the droplet properly to the specified location through the wire. It is. In this method using shape memory alloy, unlike the method using the previous hook, there is no need to install a protrusion on the plate side, and there are few problems of contamination of pliers when using means such as pulling with pliers. There are advantages.

  FIG. 9 is a schematic diagram of an optical measuring apparatus according to the fourth embodiment of the present invention. This embodiment is an example in which a spiral droplet holder is also used as a spectroscopic cell. Here, an example in which the present invention is applied to an absorptiometer is shown, but the target analyzer is not limited to this. FIG. 9 (a) shows a schematic diagram of the main part of an absorptiometer that is the fourth embodiment of the present invention. The light emitted from the lamp 501 passes through the inside of the spiral droplet holder 503 that holds the target droplet solution 504 by the mirror 502. The length of the droplet holder 503 defines the optical path length L for absorption measurement, and the support base 505 for adjusting the distance can change the distance and rotate. Although the support 505 is not essential for the absorbance measurement itself, it is important as an operation requirement in the absorbance measurement in a minute volume. The light passing through the droplet-like solution 504 is wavelength-dispersed by the diffraction grating 506 and then received by the photomultiplier tube 507 connected to the data processor 508. With this configuration, the absorbance of the solution can be measured. In an absorptiometer, since the solution is usually put into a cell made of quartz or plastic and the upper part is opened, the optical path of the light absorption part is often substantially parallel to the horizontal direction. Even in this configuration, the optical path is substantially parallel to the horizontal direction, and as a result, the spiral axis of the droplet holder is set to be substantially in the horizontal direction, but the droplet 504 is stably held even in such a state. The

According to this method, it is possible to easily perform absorption measurement of a minute amount of liquid, which has been difficult in the past. It is also possible to collect the measured solution and dispose of the spectroscopic cell. For example, consider the case of performing an absorption analysis with a liquid volume of 1 microliter. The inner diameter of the helix is d, the length of the helix is L (same as the optical path length), the number of turns is n, and the length of the wire constituting the helix is l. Considering a spiral of a stainless steel 125-micron wire wound 10 times so that the inner diameter of the wire is 1 mm, L = 125 microns x n (= 10) = 1.25 mm, V = (πd ^{2/4) × L = (} 3.14 × 1 [mm] 2 /4)×1.25[mm]=0.98[mm 3] , and the internal volume is approximately 1 microliter. For example, considering the absorbance measurement of nucleic acids and proteins where recovery of the solution is important, an aqueous solution may be assumed, and if a hydrophilic metal wire is used, it can be held well. Of course, it is possible to modify the surface of a resin wire, and in this case, since the wire is in close contact, there is virtually no problem. The length l of the wire is approximately l = πdn = 3.14 × 1 mm × 10 = 31.4 mm. Since only a very short wire is required in this way, it can be disposable as an inexpensive spectroscopic cell, and absorption measurement without worrying about contamination can be performed. If the concentration of the molecule to be measured in the solution is sufficient, the absorbance can be measured sufficiently with an optical path length of 1.25 mm, but if the concentration is low, it is necessary to increase the optical path length. In that case, it is possible to extend the optical path length from 1.25 mm to 5 mm by operating the support base 505. In order to maintain the diameter of the helix so as not to change the internal volume, it can be understood from the calculation that the number n of turns should be changed from 10 to 20 by using the support base 505. At this time, the diameter of the spiral is about 500 microns, and the distance between the wires is about 250 microns. The solution to be measured is kept sufficiently stable during the measurement time. By using a thinner wire and forming a spiral with a smaller diameter, it is possible to measure absorbance with a smaller amount of liquid. Although the optical path length is slightly shorter than the length of the spiral droplet holder due to the meniscus on the surface of the droplet, this optical path length error is not a problem in general absorption measurement. It is also possible to ensure an accurate optical path length by designing the droplet composition in consideration of the dent amount. Further, as shown in FIG. 9B, an accurate optical path can be obtained by placing a substantially transparent member 509 such as glass on both sides of the spiral droplet holder and forming a liquid bridge between the members. It is also possible to ensure the length. A method of placing a flat member is also effective, and a method of placing a member having the effect of a lens instead so that the light from the outside is collected and efficiently passes through the inside of the droplet is also effective. is there.

  FIG. 10 is a schematic diagram relating to a droplet recovery method according to a fourth embodiment of the present invention. With respect to the droplet 504 held in the droplet holder 503, the protrusion 512 coming out of the container 511 is hooked on the substantial center of the spiral droplet holder 503 or a portion near the center. By pulling the protrusion 512, the spiral droplet holder 503 is deformed, and the droplet 504 falls into the container 511 and can be recovered as the solution 513. Thus, the direction of deformation need not be in the direction of the axis of the helix. It is desirable that the protrusion 512 is, for example, linear and the angle does not deviate much from the vertical so that liquid can be easily transmitted. This collection method can be incorporated into the spectrophotometer device, or the droplet holder can be taken out of the device and operated by hand. By using this liquid holding part as a spectroscopic cell, the solution after absorption measurement can be easily recovered. In the case of the current absorption measurement using a quartz cell, etc., the solution is collected by a thin liquid supply means, etc., but the surface area that comes into contact is large, and the liquid remains in the corner (dent) inside the cell, so all It is difficult to collect. According to the present invention, there is an advantage that almost the entire amount can be recovered.

  FIG. 11 is a schematic diagram relating to a method for generating a plurality of droplets according to the fifth embodiment of the present invention. A droplet generating member 602 including a plurality of spiral droplet holders 601 and a moving means for connecting them as shown in FIG. 11A is used with a handle 606 as shown in FIG. And put on the container 603 containing the solution to be made into droplets. The connection of the droplet holder 601 is possible, for example, from about 2 to about 10,000. The entire droplet generating member can be bent and the entire can be placed in a small container 603. When the droplet generation member 602 attached to the container 603 is pulled up, the droplets are held in the respective droplet holders 601. By pulling at both ends of the droplet generating member, as shown in FIG. 11 (c), by using at least one of the both ends of the droplet generating member to deform the droplet holding portion, a plurality of the droplet generating members are pulled. Droplets 604 are generated on the substrate 605 all at once. Since the original droplet holder 601 has a simple structure and can be made uniformly and inexpensively, it can be used as an initial step for producing uniform particles, for example.

  FIG. 12 is a configuration example of a system capable of simultaneously executing a plurality of series of operations of holding, transporting, dropping and mixing droplets in the sixth embodiment of the present invention. In this embodiment, as shown in FIG. 12, when there are a plurality of wells 702 on the microplate 701, the solution 703 in the well 702 can be moved to another well 702, and a plurality of droplet holders An example of a system that can simultaneously perform solution transfer between wells 702 using 401 simultaneously is shown. In this example, the movement of the solution from the container containing the solution outside the microplate 701 to the well 702 on the microplate 701 and vice versa are not described, but it is obvious that the solution can be executed in the same configuration.

  In this embodiment, the solution is moved between containers using a spiral droplet holder 401 made of the shape memory alloy described in the third embodiment. Therefore, a mechanism for moving the droplet holder 401 on the microplate is provided. As shown in FIG. 12, the droplet holding unit 401 is connected to the driving units 706 and 707 via a handle 403. The driving units 706 and 707 can move along the groove 705 on the driving unit holding substrate 704. The groove 705 penetrates the driving unit holding substrate 704, and when the driving unit 706 or 707 moves on the substrate 704, the droplet holding unit 401 can move while being positioned directly below the driving unit. . The drive unit 706 or 707 has a spherical wheel 709 for moving on the substrate, and is movable in both the x and y directions. In the present embodiment, the moving method using the wheels is described, but the driving unit may be moved by magnetic force or electric force by arranging magnets or electrodes on the driving unit holding substrate. In addition, wheels 708 for moving the droplet holding unit 401 in the z direction are provided in the driving units 706 and 707. By rotating the wheel 708, the handle 403 moves up and down in the z direction. Further, the heater 404 is provided to drop the liquid droplet held in the well 702 by changing the pitch of the spiral liquid droplet holding portion, and preferably by forming a shape on a line. Since the heater 404 is for controlling the temperature of the spiral droplet holder 401, a Peltier element or the like capable of increasing and decreasing the temperature may be used.

  Next, a method of moving the droplet will be described. First, the driving unit 707 is moved onto the container with the solution, and the holding unit 401 is moved by driving the wheel 708 in a state where the heater is turned off, that is, the pitch of the spiral droplet holding unit 401 is not widened. Lower to the bottom. When the droplet holder 401 is lowered to the bottom of the container so that the surface of the spiral droplet holder and the solution are in sufficient contact, the liquid moves into the droplet holder. Thereafter, the wheel 708 is driven to lift the droplet holding unit 401 from the microplate 701, and the driving unit 707 is moved directly above the target well by the spherical wheel 709. After the movement in the xy direction is finished, the liquid droplet holding unit 401 is moved downward again, and then the pitch of the helical liquid droplet holding unit 401 made of a shape memory alloy is expanded using a heater to hold and hold the liquid. A drop is dropped into the well 702. The driving unit 706 shows the driving unit moving in the xy direction, and 707 shows the state of the driving unit immediately after the completion of droplet collection. In addition, the solution can be mixed by dropping the droplets on a container in which the solution already exists.

  Next, control of droplet operation will be described. In this embodiment, a plurality of driving units (droplet holding units) are controlled by the controller 710, and a plurality of droplet operations, that is, movement and mixing of solutions can be executed simultaneously. Since there is no upper limit on the number of driving units (droplet holding units) in principle, in this embodiment, complex solution operations can be performed easily and in parallel. In FIG. 12, a drive signal line 712 transmits a signal for moving the droplet holder in the xyz direction from the controller to the drive unit 706 or 707, and a temperature control line 711 supplies a current to the heater. In this case, the signal line may be a coated metal wiring or the like, but it goes without saying that wireless signal transmission and power transmission may be performed. Further, in order to stabilize the driving unit holding unit, two holding substrates 704 may be provided and moved with the driving unit interposed therebetween. Furthermore, there may be a plurality of microplates 701, and they can be removed freely. In addition, it is easy to remove, clean and replace the droplet holder.

  FIG. 13 is a schematic diagram relating to the main part of the blood biochemical analyzer of the sixth embodiment of the present invention. FIG. 13A is a schematic diagram of the apparatus configuration. At the center is a sample circulation device 801 that suspends a number of sample reaction jigs that function as sample holding means, which will be described later, and circulates each part, and a device in charge of each part is disposed around the sample circulation device 801. Blood, which is a specimen, is held in a test tube on a sample tray by a sample management device 802 that functions as a sample storage unit. The amount of liquid corresponding to each inspection is collected from each specimen and held in the sample reaction jig for the number of inspection items. The sample reaction jig is circulated by the sample circulation device 801, and after a reagent corresponding to each inspection item is added by the reagent management device 803 functioning as a reagent supply means, the detection unit 804 measures the reaction result. After the measurement, the sample reaction jig discards the waste liquid with the waste liquid discarding apparatus 805, and then is washed with the reaction jig cleaning apparatus 806, and used for the measurement of the next blood sample.

  FIG. 13B is an enlarged view of a part of the sample circulating device 801. The sample circulation device 801 is a device that suspends and circulates the sample reaction jig 811. The sample reaction jig 811 is a jig composed of the droplet holding portion and its periphery described in the above embodiment, and can hold a liquid. Here, a sample reaction jig 811 having a spiral droplet holding part and dropping protrusions is shown. The sample reaction jig 811 hangs on the circulation rail 815 via the reaction jig fixing portion 812 and the rail fixing jig 814, and moves along the circulation rail 815. The reaction jig fixing part 812 and the rail fixing jig 814 are connected by an elevating rope 813, and the sample reaction jig 811 can be operated in the vertical direction by extending this part.

  FIG. 13C is a schematic diagram in which a part of the sample management device 802 is enlarged. A number of test tubes 822 containing blood are held on the sample tray 821. The sample tray 821 operates with respect to the sample reaction jig 811, and a test tube containing a target specimen is arranged directly below the sample reaction jig 811. By moving the sample reaction jig 811 up and down by the sample circulation device 801, a predetermined amount of blood can be held in the sample reaction jig 811. Here, when the sample reaction jig 811 was attached to the test tube, the blood was held only in a part of the droplet holder by controlling the height of the sample reaction jig 811. In this example, only one test tube and one sample reaction jig are illustrated, or in practice, a predetermined number of sample tubes are held from a large number of test tubes according to the number of test items. become. The retained blood 823 is sequentially transferred to the reagent management device 803 by the sample circulating device 801.

  FIGS. 13D and 13E are schematic diagrams showing how the reagent management unit 803 adds the reagent 827 to the blood 823. FIG. The reagent 827 may be held in the reagent holding jig 826 by forming the structure shown in FIG. 5 as an apparatus. When the reagent holding jig 826 holding the reagent 827 is operated, the sample circulating apparatus 801 is brought close to the sample reaction jig 811 holding the blood 823 transferred to the reagent management unit 803, and the liquids are brought into contact with each other. By releasing the tool 826, the reagent 827 can be transferred to the sample reaction tool 811 and mixed with the blood 823. In the mixed blood / reagent mixed solution 828, a chemical reaction according to the inspection item proceeds, and is inspected by the next inspection unit 804. Here, only one sample reaction jig of one kind of reagent is described, but actually, the reagent is mixed according to the number of items and the reaction proceeds.

  FIG. 13 (f) shows a schematic diagram of the inspection section. Here, the description is made with absorption measurement in mind, but the measurement method is not limited to this, and there are fluorescence, luminescence method, scattering, etc. as light measurement. Light emitted from the light source 831 is reflected by the mirrors 832 and 833 along the optical path 835 and then received by the photomultiplier tube 834. On the way, absorption occurs at a wavelength according to the target inspection target item through the blood / reagent mixed solution 828 after the reaction held in the sample reaction jig 811. Therefore, the inspection can be performed by the output of the photomultiplier tube 834. it can. The solution after the inspection is sent to the waste liquid discarding device 805. FIG. 13 (g) shows a schematic diagram of the waste liquid discarding device 805. FIG. The mechanism is such that the waste liquid 843 falls into the waste container 841 by hooking the tip of the sample reaction jig 811 up and down on the protrusion 842 coming out of the waste container 841. The sample reaction jig 811 is sent to the reaction jig cleaning device 806 by the sample circulation device 801. FIG. 13 (h) is a schematic view of cleaning by the reaction jig cleaning device 806. The sample reaction jig 811 is cleaned in the cleaning tank 846. For example, it is possible to increase the cleaning effect by irradiating ultrasonic waves. The cleaned sample reaction jig 811 is also used for the next inspection. If contamination is a serious problem, it is possible to install a device that replaces the sample reaction jig with a new one without washing.

  As shown in FIGS. 13 (a) to (h), the present liquid conveying means can be applied to an automatic blood analyzer. By using a large number of inexpensive and simple sample reaction jigs 811, the structure is simple and inexpensive automatic blood analysis is possible. Even when the reaction time / reaction temperature differs for each inspection item, it can be easily handled by creating a loop and a thermostat in the sample circulation device.

  In the above embodiment, the description has been given by taking the droplet holding portion as an example of an annular and spiral droplet holding portion, but the present invention is not limited to these two shapes, and can hold a droplet. This also applies to similar structures. Among these, a combination of two or more parts to form a space for holding a droplet is also included. In addition, since the evaporation speed increases as the droplets become smaller, it is also effective to install an evaporation-prevented sealed chamber or humidification chamber around the entire apparatus or around the droplet holder if necessary.

101: droplet holding part, 102: droplet, 103: handle, 104: stirring rod, 105: stirring motor, 106: eccentric motor, 110: container, 111: liquid film, 112: droplet, 113: liquid 201: Droplet holder, 202: Droplet, 203: Handle, 211: Droplet holder, 212: Droplet, 213: Handle, 220: Mixed droplet, 221: Mixed Droplet, 222: Liquid film, 301: Droplet holder, 302: Droplet, 303: Handle, 304: Projection, 310: Container containing solution, 311: Well of microplate, 312: Protrusion, 313 : Liquid, 401: Droplet holder, 402: Droplet, 403: Handle, 404: Heater, 405: Controller, 406: Test tube, 407: Linear wire, 408: Liquid, 501: Lamp, 502: Mirror: 503: Droplet holder 504: Solution, 505: Support base, 506: Diffraction grating, 507: Photomultiplier tube, 508: Data processing device, 509: Transparent member, 511: Container, 512: Projection, 513: Solution, 601: Droplet holder 602: Member for generating droplets, 603: Container, 604: Droplet, 605: Substrate, 606: Handle, 607: Member deformation means, 701: Microplate, 702: Well, 703: Solution, 704: Holding unit Substrate, 705: groove, 706: drive unit, 707: drive unit, 708: wheel, 709: wheel, 710: controller, 711 temperature control line, 712: drive signal line 801: sample circulation device, 802: sample management device, 803: Reagent management device, 804: Detection unit, 805: Waste liquid discarding device, 806: Reaction jig cleaning device, 811: Sample reaction jig, 812: Reaction jig fixing unit, 813: Lifting rope, 814: Rail fixing jig, 815: Rail, 821: Sample tray, 822: Test tube, 823: Blood, 826: Reagent holding jig, 827: Reagent, 828: Mixed solution, 831: Light source, 832: Mirror, 833: Mirror, 834: Photomultiplier tube, 835: optical path, 841: waste container, 842: protrusion, 843: waste liquid, 846: washing tank.

Claims (6)

A droplet holding portion made of a helical wire made of a shape memory alloy;
A light irradiation means for irradiating light inside the spiral of the droplet holding portion;
And a detecting means for detecting light that has passed through the inside of the spiral.   The photodetection device according to claim 1, further comprising a plurality of substantially transparent members that are respectively installed so as to sandwich the droplet holding portion.   The container further includes a container for collecting the droplet and a container protrusion disposed on the container, and the container protrusion is hooked on a substantial center of the droplet holding portion or a portion near the center thereof. The photodetecting device according to claim 1, wherein the holding portion is deformed. A storage unit for storing the sample;
Sample holding means for collecting and holding a part of the sample from the sample storage unit;
A circulating part for circulating the sample holding means;
A reagent supply unit for supplying a reagent to the sample holding means circulated by the circulation unit;
A detection unit configured to detect the sample held by the sample holding unit, and the sample holding unit includes a first droplet holding portion and a first moving unit for inscribed the droplet of the sample. The analyzer is characterized in that the reagent supply unit includes a second droplet holding portion for inscribed the reagent droplets and a second moving means.   5. The analyzer according to 4, wherein the first droplet holding portion is made of an annular wire, and the second droplet holding portion is made of a helical wire. Holding each of the droplets in a plurality of droplet holding portions that inscribe the droplets; and
Bringing the droplets held in each of the plurality of droplet holding portions into contact;
Mixing the droplets by the contact;
In the holding step, at least two droplet holding portions having different diameters are used, and in the mixing step, the droplet moves to the droplet holding portion having a larger diameter. Mixing method.

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