KR101929414B1 - Microfluidic dilution device and method for dilution using the same - Google Patents

Abstract

A body having a flow path through which a fluid is fed and a valve for selectively opening and closing the flow path so as to control the flow of the fluid so that the dilution concentration of the sample can be variously adjusted by continuously diluting the sample at an arbitrary ratio in one apparatus; A buffer solution supply part for supplying the buffer solution, a buffer part for supplying the buffer solution, a buffer part for supplying the buffer solution to the sample part, and a storage part for storing the diluted solution, And a sample metering unit connected to the mixing chamber for discharging the solution in a predetermined amount and for retaining the metered solution in the mixing chamber.

Description

TECHNICAL FIELD [0001] The present invention relates to a microfluidic dilution device and a dilution method using the microfluidic dilution device.

Disclosed is a microfluidic dilution device for diluting an object sample at an arbitrary ratio through a fluid flow and a fluid dilution method using the same.

Continuous dilution of the sample is an essential process in biochemical reaction experiments. In order to produce diluted samples in various stages, complicated pipetting process and time, manpower and other consuming tasks must be accompanied.

Biochemical reaction experiments that frequently utilize microfluidic solutions of several micrometers require more precise and accurate serial dilution. In addition, there is a demand for a technique capable of adjusting the dilution concentration at an arbitrary ratio other than a specific dilution ratio in accordance with various experimental conditions.

The present invention provides a microfluidic dilution device and a dilution method using the microfluidic dilution device, wherein the dilution concentration of the sample can be varied by continuously diluting the sample in an arbitrary ratio in one device.

The present invention also provides a microfluidic dilution device which can be easily manufactured with a simpler structure and can control the sample dilution concentration more easily and easily, and a dilution method using the same.

The microfluidic dilution device of this embodiment includes a body having a flow path through which a fluid is transferred and a valve for selectively opening and closing the flow path to control the flow of the fluid, a diluting part formed along the flow path to dilute the sample, A buffer chamber for supplying a buffer solution; a mixing chamber connected to the buffer solution supply unit for mixing and diluting the buffer solution with the sample solution; And a sample metering section connected to the mixing chamber for discharging the solution from the mixing chamber in a fixed amount and for retaining the metered solution in the mixing chamber.

And a sample supply unit connected to the mixing chamber and capable of supplying the sample and transferring the supplied sample to the mixing chamber.

And a buffer solution metering unit connected between the buffer solution supply unit and the mixing chamber for metering and supplying the buffer solution.

Wherein the buffer solution metering unit comprises at least one or more buffer solution metering chambers disposed on the outward side of the buffer solution supply unit for metering and storing a buffer solution, an open / close valve provided on the out side of the buffer solution supply unit, And a supply valve installed in the supply passage formed in the support member and opening and closing the supply passage.

The buffer solution metering section is connected to the mixing chamber with at least one or more buffer solution metering chambers having a volume corresponding to the set dilution magnification, and a supply valve is provided between the buffer solution metering chamber and the supply channel or the adjacent buffer solution metering chambers, The buffer solution metering section may have the same volume as each of the buffer solution metering chambers, or at least one of the metering chambers may be formed in a different volume.

The buffer solution metering unit further includes a waste flow channel connected to the buffer solution metering chamber and flowing excess fluid exceeding the volume of the metering chamber, and a waste chamber connected to the waste flow channel to receive excess solution discarded through the waste flow channel .

The sample metering section includes a metering channel connected to the outflow side of the mixing chamber for discharging the solution, a discharge valve for opening and closing the outflow side of the mixing chamber, and a metering channel provided at intervals along the metering channel to open and close the metering channel, And at least one metering valve for dividing the metering valve into the volume section.

The volume of the metering channel may be the same or the volume of at least one volume may be different.

The metering channel may have a single channel structure having an entire internal cross-sectional area.

The storage unit may include at least one storage chamber connected to the diluter outlet for receiving the diluted solution transferred through the diluter, and a storage valve for opening and closing the storage chamber inlet.

The storage unit may have a structure in which a plurality of storage chambers are sequentially connected along a discharge flow path connected to a diluter outlet and a shutoff valve is installed in a discharge flow path between the respective storage chambers.

The body may have a rotation center to apply a centrifugal force to the fluid to transfer the fluid along the flow path.

The buffer solution supply unit, the buffer solution metering unit, the mixing chamber, the sample metering unit, and the storage unit may be sequentially arranged outward from the center of the body and connected in series.

And an air hole formed in the body and communicating with the flow path to the outside.

In the dilution method of this embodiment, a sample and a buffer solution are supplied to a main body, a sample contained in the mixing chamber of the main body is discharged in a predetermined amount, the sample is left in the mixing chamber, the buffer solution is transferred to the mixing chamber, Mixing and diluting, and discharging and storing the diluting solution in the mixing chamber.

Transferring the buffer solution to at least one or more metering chambers prior to transferring the buffer solution to the mixing chamber, and transferring the buffer solution of at least one metering chamber to the mixing chamber according to the dilution ratio.

And transferring the buffer solution to the weighing chamber and removing the excess solution in each weighing chamber when weighing.

When the dilution liquid is discharged from the mixing chamber, the diluted liquid in the mixing chamber is discharged in a predetermined amount, the diluted liquid measured in the mixing chamber is remained, and the buffer solution is transferred to the mixing chamber to mix and dilute the diluted solution. And repeating the above steps to continuously dilute the solution.

The continuous dilution step may be repeatedly carried out in the same sample metering volume and buffer solution volume and serially diluted in the same ratio.

The continuous dilution may be repeatedly carried out at different sample metering volumes and buffer solution volumes and serially diluted in any proportion.

As described above, according to this embodiment, the sample can be continuously diluted at an arbitrary ratio.

It becomes possible to more easily adjust the dilution concentration in one apparatus.

A more accurate and accurate dilution ratio can be obtained through metering.

It is easier to manufacture with a simpler structure, and sample dilution concentration can be easily and easily adjusted.

1 is a schematic view showing a microfluidic dilution device according to the present embodiment.
FIG. 2 and FIG. 3 are views showing a part of the configuration of the microfluidic dilution apparatus according to the present embodiment in more detail.
4 is a schematic cross-sectional view showing the valve structure of the microfluidic dilution device according to the present embodiment.
5 and 6 are schematic views for explaining a sample dilution process of the microfluidic dilution apparatus according to the present embodiment.
FIG. 7 is a graph showing the results of microfluidic dilution according to this embodiment.

Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings so that those skilled in the art can easily carry out the present invention. It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention. Wherever possible, the same or similar parts are denoted using the same reference numerals in the drawings.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the invention. The singular forms as used herein include plural forms as long as the phrases do not expressly express the opposite meaning thereto. Means that a particular feature, region, integer, step, operation, element and / or component is specified, and that other specific features, regions, integers, steps, operations, elements, components, and / And the like.

All terms including technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention belongs. Predefined terms are further interpreted as having a meaning consistent with the relevant technical literature and the present disclosure, and are not to be construed as ideal or very formal meanings unless defined otherwise.

FIG. 1 shows a microfluidic dilution device according to the present embodiment, and FIGS. 2 and 3 show a detailed configuration of a part of a microfluidic dilution device according to the present embodiment.

The microfluidic dilution device of this embodiment will be described with reference to Figs. 1 to 3. Fig.

The microfluidic dilution device 100 of the present embodiment includes a body 10 having a flow path through which a fluid is transferred and a valve for selectively opening and closing the flow path to control the flow of the fluid, And a storage part (70) connected to the diluting part and accommodating the diluting solution.

The main body may be connected to the mixing chamber 24 and may be provided with a sample supply unit 20 capable of supplying the sample and transferring the supplied sample to the mixing chamber.

The diluter continuously dilutes the sample at a dilution ratio according to an arbitrary magnification. The dilution unit includes a buffer solution supply unit 22 for supplying a buffer solution, a mixing chamber 24 connected to the buffer solution supply unit 22 to dilute and mix the sample and the buffer solution, And a sample metering section 50 for discharging the solution in the metered amount and retaining the metered solution in the mixing chamber 24. The dilution unit may further include a buffer solution metering unit 30 connected between the buffer solution supply unit 22 and the mixing chamber 24 for metering the buffer solution to supply the buffer solution to an accurate capacity.

In the following description, a sample refers to a fluid containing a sample, a buffer solution means a solution for diluting a sample, and a diluent means a solution in which a sample and a buffer solution are mixed and diluted. Fluid or solution may refer to a sample, buffer solution or diluent.

The body 10 may be, for example, in the form of a rotatable circular disk. The center of the body 10 can be rotated about the rotation axis by a driving force provided from the outside in the form of a rotation axis. A centrifugal force is generated in accordance with the rotation of the body 10, and the fluid inside the body 10 is conveyed by the centrifugal force. A flow path is formed so as to allow the microfluid to flow into the body 10. The flow path may be formed in a direction from the center of the body 10 toward the outer end so that the fluid can flow by the centrifugal force applied to the body 10. [ And each constituent part of the dilution part is sequentially disposed along the flow path and connected to the flow path. When the centrifugal force is applied to the body 10, the fluid flows from the sample supply unit 20 and the buffer liquid supply unit 22 to the mixing chamber 24, the sample metering unit, and the storage unit 70. Accordingly, as the centrifugal force is applied by rotating the body 10, the fluid can be sequentially moved to perform a desired process.

A valve for opening and closing the flow path as needed is provided at one side of the flow path. As the valve is driven, the fluid moves or is blocked along the flow path to control the flow of the fluid.

FIG. 4 shows an embodiment of a valve provided in the body. As shown in FIG. 4, the valve 80 may include an elastic plate 81 installed on the body for selectively blocking the flow passage, and a pressing portion provided on the body for pressing the elastic plate 81. The elastic plate 81 is provided on the flow path, and the pressing portion is provided at a position corresponding to the elastic plate on the outside of the body. The elastic plate 81 is made of a material having elasticity itself. The elastic plate 81 is pressed by an external force to cut off the flow path, and when the external force is removed, the elastic plate 81 returns to the original state and opens the flow path. The pressing portion includes a housing 82 mounted on the body, a pressing bar 83 which is movably installed in the housing and which presses the elastic plate 81, and a pressing bar 83 for fixing or releasing the pressing state of the pressing bar And a latching protrusion 85 provided on the housing 82 for latching the latching protrusion 84 therebetween.

When the actuating force is externally applied to press the push bar 83 of the valve 80 and the engaging protrusion 84 is fixed to the engaging protrusion 85, the push bar is fixed in a pressed state. On the contrary, if the locking protrusion 84 is removed from the locking protrusion 85, the pushing bar 83 is moved to the original position by the elastic force of the elastic plate 81. The elastic plate 81 is pushed by the movement of the push bar 83 to block the flow path or return the elastic plate 81 to the original position to open the flow path. Various structures other than the above-described structure can be applied as long as the valve 80 can open and close the flow path.

In this embodiment, the valve opens and closes the flow path to regulate the capacity of the fluid for sample dilution. Therefore, the valve must be able to be opened and closed as well as installed at an appropriate position on the flow path.

Hereinafter, the structure of the diluting section for diluting the sample at an arbitrary ratio will be described.

The buffer solution supply portion 22, the buffer solution metering portion 30, the mixing chamber 24, the sample metering portion 50, and the storage portion 70 constituting the diluting portion constitute one unit. A plurality of units may be formed on the body 10 along the circumferential direction of the body 10. Thus, a dilution operation can be performed on a plurality of samples with one body 10. Each of the units formed in the body 10 may have the same structure.

1, the body 10 is provided with a sample supply unit 20, a buffer solution supply unit 22, a buffer solution metering unit 30, a mixing chamber 24, A metering section 50 and a storage section 70 are formed.

The sample supply unit 20, the buffer liquid supply unit 22, the buffer liquid metering unit 30, the mixing chamber 24, the sample metering unit 50, and the storage unit 70 in the one- And may be connected in series. Thus, the present apparatus is easy to manufacture with a simple flow path structure connected in series, and the sample can be accurately diluted and stored at an arbitrary ratio even through a simple structure.

A sample supply unit 20 and a buffer solution supply unit 22 are disposed on the entrance side of the mixing chamber 24 facing the rotation center of the body 10 along the centrifugal direction of the body 10 to connect the mixing chamber 24 do. And the sample metering section 50 is connected to the outgoing side of the mixing chamber 24 facing toward the outer tip end of the body 10 in the opposite direction.

In this embodiment, the buffer liquid supply portion 22 is connected to the mixing chamber 24 through the buffer liquid supply portion 22. [ A storage section (70) is connected to an exit side of the sample metering section (50).

In the following description, the term " inlet " means a portion into which the fluid flows into the chamber, which means a portion near the rotation center of the body 10 along the direction of centrifugal force, and " Means a side near the outer end of the body 10.

The sample solution supply unit 20, the buffer solution supply unit 22, the buffer solution metering unit 30, the mixing chamber 24, the sample metering unit 50, and the sample solution supply unit 20 are disposed along the centrifugal force direction at the center of rotation of the body 10. [ And a storage unit 70 are sequentially arranged. Therefore, the sample and the buffer solution are mixed and diluted in the mixing chamber 24 due to the centrifugal force generated when the body 10 rotates, and the diluted solution is sequentially transferred to the storage part 70 and stored.

The sample supply unit 20 supplies the sample fluid to the mixing chamber 24. The sample supply part 20 may be formed in the form of a chamber in the body 10. The chamber can be understood as an empty space formed inside the body 10. The chamber may be provided with a hole for injecting a fluid as a sample.

The chamber of the sample supply unit 20 and the mixing chamber 24 are sequentially arranged from the center of rotation of the body 10 toward the outer tip along the centrifugal force direction of the body 10 so that the fluid as a sample flows from the chamber of the sample supply unit to the mixing chamber (24).

The mixing chamber 24 is a space in which the sample and the buffer solution are mixed and diluted at a constant ratio. The sample introduced into the mixing chamber 24 by the centrifugal force applied to the body 10 and the buffer solution are mixed and diluted.

The buffer solution supply unit 22 supplies a buffer solution for sample dilution to the mixing chamber 24. [ The buffer solution supply unit 22 may be formed in the body 10 in the form of a chamber. The chamber can be understood as an empty space formed inside the body 10. A hole for injecting a buffer solution may be formed in the chamber.

The chamber of the buffer solution supply section 22 and the mixing chamber 24 are sequentially arranged from the center of rotation of the body 10 toward the outer extremity along the centrifugal direction of the body 10 so that the buffer solution is supplied from the chamber to the mixing chamber 24, And flows to the inside.

The buffer solution metering section 30 is installed between the outlet of the buffer solution supply section 22 and the inlet of the mixing chamber 24 to control the volume of the buffer solution flowing into the mixing chamber 24.

The buffer solution metering section 30 includes at least one or more buffer solution metering chambers disposed on the outward side of the buffer solution supply section 22 for metering the buffer solution and accommodating the buffer solution, an open / close valve 34 provided on the out side of the buffer solution supply section 22, And a supply valve installed in the supply passage 35 formed between the respective outlets of the buffer solution metering chambers and the mixing chamber 24 for opening and closing the buffer solution supply passage 35.

In the case of a structure in which a plurality of buffer solution metering chambers of the buffer solution metering section 30 are provided, each of the buffer solution metering chambers may be formed to have the same volume with each other or at least one of the metering chambers may be formed in a different volume have.

An on-off valve 34 for opening and closing the flow path is provided on the outflow channel of the buffer solution supply unit 22. [ When the open / close valve 34 is operated to open the outflow channel of the buffer solution supply section 22, the buffer solution is transferred to the buffer solution supply section 22 through the flow path.

Fig. 1 shows, by way of example, a structure in this embodiment in which the buffer solution metering chambers are made up of three chambers having mutually different volumes. The buffer solution metering chamber may be formed in a number of not more than two, or four or more, in addition to the structure of three chambers. The three buffer solution metering chambers may each be formed to have a volume of 5 mu L, 3 mu L, and 1 mu L volume. By providing three measuring chambers of 5 mu L, 3 mu L and 1 mu L volume in this way, it is possible to provide a more diversified dilution ratio while simplifying the structure. For convenience of the following description, a chamber of 5 占 용 volume in each of the buffer solution metering chambers is referred to as a first metering chamber 31, a chamber of 3 占 용 volume is referred to as a second metering chamber 32, It is referred to as a metering chamber 33. The buffer liquid metering chamber may refer to all three metering chambers. A supply valve is provided on the outlet side of each buffer solution metering chamber. The supply valve provided on the out side of the first metering chamber 31 is referred to as a first supply valve 36 and the supply valve provided on the out side of the second metering chamber 32 is referred to as a second supply valve 37, And the supply valve provided on the outlet side of the third supply valve 38 is referred to as a third supply valve 38. [ A supply valve may refer to all three supply valves.

Each buffer solution metering chamber may be individually connected to the mixing chamber via a respective supply line. In such a case, the metered solutions of the respective buffer solution metering chambers can be separately supplied to the mixing chamber 24 by controlling the supply valves provided in the outgoing supply channels of the respective buffer solution metering chambers.

In the present embodiment, each buffer solution metering chamber is connected to the mixing chamber 24 through one supply flow path 35. To this end, as shown in FIG. 2, each of the buffer solution metering chambers having different volumes is arranged in parallel in the buffer solution supply part 22 and sequentially connected to the mixing chamber 24. The respective inlet sides of the first metering chamber 31, the second metering chamber 32 and the third metering chamber 33 arranged in this manner are connected to the outflow side of the buffer solution supply unit 22. A common flow path 39 is extended along the entrance side of each buffer solution metering chamber arranged in parallel and the buffer solution moved along the common flow path 39 is supplied to each buffer solution metering chamber Filled in turn. The outlets of the first metering chamber 31, the second metering chamber 32 and the third metering chamber 33 are connected to the supply chamber 35 and connected to the mixing chamber 24. A first supply valve 36 is provided between the mixing chamber 24 and the first metering chamber 31 on the supply flow path 35. The first metering chamber 31 and the second metering chamber 32, And a second supply valve 37 and a third supply valve 38 are provided between the second metering chamber 32 and the third metering chamber 33, respectively.

Accordingly, the solution metered in the first metering chamber 31, the solution metered in the second metering chamber 32, and the solution metered in the third metering chamber 33 are sequentially supplied to the mixing chamber (24).

A first supply valve 36 for opening and closing the flow path is provided on the outgoing supply flow path 35 of the first metering chamber 31. When the first supply valve 36 is operated to open the supply passage 35, the metered buffer solution of the first metering chamber 31 is transferred to the mixing chamber 24 through the supply passage 35.

A second supply valve 37 for opening and closing the flow path is provided on the flow path connecting the second metering chamber 32 and the first metering chamber 31. [ When the second supply valve 37 is operated to open the flow path, the metered buffer solution in the second metering chamber 32 passes through the first metering chamber 31 and through the supply flow path 35 to the mixing chamber 24 .

A third supply valve 38 for opening and closing the flow path is provided on the flow path connecting the third metering chamber 33 and the second metering chamber 32. When the third supply valve 38 is operated to open the flow path, the metered buffer solution in the third metering chamber 33 passes through the second metering chamber 32 and the first metering chamber 31, 35 to the mixing chamber 24.

After filling each buffer solution metering chamber, excess buffer solution is discarded. To this end, a waste flow path 40 is connected to the outlet side of the common flow path 39 extending from the buffer solution supply section 22, and an excess solution discharged through the waste flow path 40 is accommodated A waste chamber 41 is connected.

Thus, it is possible to accurately fill the buffer solution of the correct capacity in each buffer solution metering chamber and to dispose of the remaining portion exceeding the volume of each metering chamber in the waste chamber 41 for disposal. A waste valve 42 for opening and closing the waste flow path 40 may be installed at one side of the waste flow path 40 connected to the waste chamber 41. The waste valve 420 is opened and closed to be discarded into the waste chamber 41 to prevent the received buffer solution from flowing into the metering chamber.

The air flow path 43 is connected to the common flow path 39 extending from the waste flow path 40 or the buffer liquid supply unit 22 and the air flow path 43 is formed with an air hole 44 communicating with the air flow path . The air passage 43 extends toward the rotation center of the body 10 to prevent the fluid conveyed according to the centrifugal force from flowing out through the air hole 44 formed in the air passage. External air flows through the air passage formed in the body 10 through the air hole 44, so that the resistance of the fluid is minimized and the fluid is smoothly conveyed through the passage.

In this embodiment, the buffer solution metering section 30 can be arranged such that a plurality of buffer solution metering chambers gradually increase in volume toward the mixing chamber 24. That is, as shown in FIG. 2, the first metering chamber 31, the second metering chamber 32, and the third metering chamber 33 are sequentially disposed from the mixing chamber 24 and connected. Accordingly, a large amount of the buffer solution in the metered buffer solution is supplied, and the dilution ratio desired can be adjusted while gradually decreasing the amount of the buffer solution supplied.

The sample metering section 50 is connected to the outflow side of the mixing chamber 24 to control the volume of the fluid remaining in the mixing chamber 24. That is, the sample metering section 50 adjusts the fluid capacity remaining in the mixing chamber 24 by removing the metered volume of fluid in the mixing chamber 24.

The sample metering section 50 includes a metering channel 51 connected to the outflow side of the mixing chamber 24 for delivering the solution, a discharge valve 52 for opening and closing the outflow side of the mixing chamber 24, 51 for opening and closing the metering channel 51 and dividing the inside of the metering channel 51 into at least one or more volume sections.

In the case of a structure in which the metering channel 51 is divided into a plurality of volume sections, the volume sections may be formed to have the same volume in accordance with the dilution ratio, or at least one volume section may be formed in a different volume.

As shown in Fig. 3, in this embodiment, the metering channel 51 can be elongated in a zigzag shape so as to have a maximum volume in a narrow area. As described above, since the metering channel 51 is formed by a single channel, the device can be easily designed and manufactured with a simple structure. Further, as the fluid moves along a single flow path, metering and control are facilitated, enabling more precise work.

The metering channel 51 may be formed in various sizes for each volume section, or may be formed to have the same overall size. When the internal cross-sectional area of the metering channel 51 is entirely the same, it is possible to easily form the metering value of each volume section by dividing the volume section according to the length.

The division of the volume section along the metering channel 51 is made by the metering valve. A metering valve is provided between each volume section along the metering channel (51).

Fig. 1 shows, as an example, a structure in which the metering channels 51 are divided into three volume sections having different volumes. The metering channel 51 may have a volume structure of two or less, or four or more, in addition to the structure of three volume sections. The three volume intervals may be formed to have a volume of 5 mu L, 3 mu L, and 1 mu L, respectively. By providing three volume intervals of 5 占 퐇, 3 占 퐇 and 1 占 퐇 volume in this way, it is possible to provide a more diversified dilution ratio while simplifying the structure. For convenience of the following description, a volume interval of 5 占 용 volume is referred to as a first volume interval 53, a volume interval of 3 占 을 volume is referred to as a second volume interval 54, and a volume interval of 1 占 volume 3 volume section (55). The metering channel 51 or the volume section may refer to all three volume sections. The first volume section 53, the second volume section 54 and the third volume section 55 are arranged in order along the metering channel 51. [ A weighing valve is installed at each volume outlet. The metering valve provided on the exit side of the first volume section 53 is referred to as a first metering valve 56. The metering valve provided on the exit side of the second volume section 54 is referred to as a second metering valve 57, And the metering valve provided on the outlet side of the second metering valve 55 is referred to as a third metering valve 58. [ The interval between the discharge valve 52 and the first metering valve 56 along the metering channel 51 forms the first volume section 53. [ The interval between the first metering valve 56 and the second metering valve 57 forms the second volume section 54 and the interval between the second metering valve 57 and the third metering valve 58 forms the third volume section 54. [ Thereby forming the volume section 55.

When the first metering valve 56 is operated to close the metering channel 51, the fluid discharged from the mixing chamber 24 flows into the first volume section 53 of the metering channel 51, The fluid in the mixing chamber 24 can be metered and discharged only by the capacity of the section 53. [

When the metering channel 51 is opened and the second metering valve 57 is operated to close the metering channel 51 by operating the first metering valve 56, the solution is filled up to the second volume section 54, The fluid can be metered and discharged in the mixing chamber 24 by the sum of the volume of the volume section 53 and the volume of the second volume section 54.

When the metering channel 51 is opened by operating the first metering valve 56 and the second metering valve 57 and the metering channel 51 is closed by operating the third metering valve 58, The fluid is metered in the mixing chamber 24 by the sum of the capacities of the first volume section 53, the second volume section 54 and the third volume section 55 and discharged.

As the fluid is metered and discharged from the mixing chamber 24 through the sample metering section 50, the fluid can be left in the flow volume set in the mixing chamber 24.

As described above, the fluid discharged through the outflow side of the mixing chamber 24 is precisely metered as it is filled in each volume section. Thus, the flow rate of the fluid remaining in the mixing chamber 24 can be accurately measured. That is, by discharging and removing the metered amount of the flow amount of the fluid contained in the mixing chamber 24, it is possible to measure the remaining fluid amount in the mixing chamber 24 and to allow only the desired amount of fluid to remain in the mixing chamber 24 do.

In the present embodiment, the sample metering section 50 may have a structure in which the volume of each volume section gradually decreases along the metering channel 51 from the outside of the mixing chamber 24. 3, the first volume section 53, the second volume section 54, and the third volume section 55 are sequentially arranged from the outflow side of the mixing chamber 24. Accordingly, it is possible to gradually adjust the dilution ratio within the mixing chamber 24 while reducing the amount of the exhaust gas.

The storage unit 70 is connected to the diluting unit to sequentially store the diluted liquid conveyed along the diluting unit. The storage unit 70 includes at least one storage chamber connected to the outside of the sample metering unit 50 for receiving the fluid transferred from the mixing chamber 24 through the sample metering unit 50, And may include a storage valve that opens and closes.

As shown in FIG. 1, five storage chambers are sequentially arranged along a discharge passage 61 connected to the outflow side of the metering passage 51. Thus, diluent solutions having different dilution ratios can be sequentially stored in the respective storage chambers. The number of storage chambers formed can be varied.

Each of the storage chambers has a first storage chamber 62, a second storage chamber 63, a third storage chamber 64, a fourth storage chamber 65, 5 storage chamber (66). The storage chamber may refer to all five storage chambers. The storage valves provided at the inlet sides of the first storage chamber 62, the second storage chamber 63, the third storage chamber 64 and the fourth storage chamber 65 are connected to the first storage valve 67, The second storage valve 68, the third storage valve 69, and the fourth storage valve 70, respectively. A storage valve may refer to all four storage valves.

In the present embodiment, the storage unit 70 is sequentially connected to the respective storage chambers along the discharge channel 61 connected to the sample metering unit 50. A shutoff valve for shutting off the discharge flow path 61 is provided in the discharge flow path 61 between the respective storage chambers. The shut-off valve opens and closes the discharge passage 61 to sequentially transfer the fluid to the corresponding storage chamber along the discharge passage 61, and blocks the transfer to the other storage chamber. The shut-off valve may be disposed immediately behind the storage valve of each storage chamber along the discharge passage 61. The shutoff valves disposed behind the storage valves of the respective storage valves along the discharge flow path 61 are connected to the first shutoff valve 71, the second shutoff valve 72, the third shutoff valve 73 ) And a fourth shut-off valve (74). A shutoff valve may refer to all four shutoff valves.

A separate storage valve for opening and closing the flow path is not provided at the inlet side of the fourth storage chamber 66, and a fourth shut-off valve provided in the discharge flow path 61 functions as a storage valve for opening and closing the inlet.

The discharge passage 61 is connected to the inlet side of the one storage chamber, and the shutoff valve and the storage valve are operated to open and close the flow passage, whereby the fluid transferred along the discharge passage 61 is introduced into each storage chamber and sequentially stored .

Hereinafter, a process of diluting the sample through the microfluidic dilution apparatus will be described with reference to FIG. Hereinafter, the reference numerals used in the respective structural parts refer to the reference numerals shown in Fig.

Fig. 5 (a) shows a process of continuously diluting the sample at a dilution ratio of 2 times magnification.

First, a sample and a buffer solution are injected into the body 10 and prepared. For example, 10 μl of sample is injected into the sample supply part 20 of the body 10 and 150 μl of buffer solution is injected into the buffer solution supply part 22. At this time, the valves installed on the flow path of the body 10 are closed and the respective flow paths are kept closed.

When the preparations are completed, the sample is transferred to the mixing chamber 24 and the set amount of the sample contained in the mixing chamber 24 is discharged to leave only the metered sample in the mixing chamber 24.

When the body 10 is rotated, centrifugal force is applied to the sample, and the sample accommodated in the sample supply unit 20 is transferred to the mixing chamber 24. Then, the discharge valve 52 provided on the outgoing side of the mixing chamber 24 is opened and the first metering valve 56 provided in the metering channel 51 is closed. The fluid in the mixing chamber 24 is discharged to the outlet side of the mixing chamber 24 by the centrifugal force and flows along the metering channel 51. Since the first metering valve 56 of the metering channel 51 is closed and the metering channel 51 is closed, the fluid discharged from the mixing chamber 24 flows through the first volume section 53 of the metering channel 51, .

Since the volume of the first volume section 53 is preset to 5 mu l, 5 mu l of the fluid contained in the mixing chamber 24 is metered into the first volume section 53 and discharged into the mixing chamber 24 Lt; RTI ID = 0.0 > 5 < / RTI >

As described above, the fluid is transferred to the first volume section 53 of the metering channel 51, so that only a fixed amount of fluid measured in the mixing chamber 24 can be left. (See 1 in Fig. 5 (A)))

After the sample is metered and remained in the mixing chamber 24, the sample filled in the first volume section 53 of the metering channel 51 is discharged through the metering channel 51. The first metering valve 56 blocking the exit of the first volume section 53 is opened by closing the discharge valve 52 on the outgoing side of the mixing chamber 24 to shut off the discharge of the fluid, ). Thus, the sample in the first volume section 53 is transferred along the metering channel 51 and is introduced into the first reservoir chamber 62 connected to the discharge channel 61 and stored. At this time, the first shutoff valve 71 provided in the discharge flow path 61 is closed to shut off the discharge flow path 61 and the first storage valve 67 is opened so that the inlet side of the first storage chamber 62 is opened Respectively. The sample transferred to the discharge passage 61 flows into the first storage chamber 62 through the inlet of the first storage chamber 62 opened in the discharge passage 61 and is stored.

The buffer solution is transferred to the mixing chamber 24 to dilute the sample in a state in which a predetermined amount of the sample remains in the mixing chamber 24. The buffer solution may be metered before the buffer solution is transferred to the mixing chamber 24 to dilute the sample with the set dilution ratio and the metered buffer solution may be diluted by transferring it to the mixing chamber 24 according to the dilution ratio.

When the open / close valve 34 provided on the outflow side of the buffer solution supply part 22 is opened, the buffer solution flows into each buffer solution metering chamber through the common flow path. After the buffer liquid metering chambers are filled, excess buffer liquid flows out to the waste chamber 41 along the waste flow path 40 connected to the common flow path, and is removed (see 2 in Fig. 5 (A)).

Thus, each buffer solution metering chamber is metered and housed in a buffer solution having a capacity corresponding to the designed volume. In this embodiment, 5 mu l of the buffer solution is accommodated in the first metering chamber 31, 3 mu l of the buffer solution is accommodated in the second metering chamber 32, A buffer solution is received.

When the buffer solution is received in each buffer solution metering chamber, the open / close valve 34 provided on the outflow side of the buffer solution supply part 22 is closed to interrupt the supply of the buffer solution, and the waste valve installed in the waste flow path 40 is also closed Thereby shutting off the waste flow path 40.

Then, the buffer solution metered in the buffer solution metering chamber accommodating the buffer solution of the capacity in accordance with the dilution ratio is supplied to the mixing chamber 24, and the sample is diluted with the desired dilution ratio.

In this embodiment, 5 占 퐇 of the buffer solution must be mixed with 5 占 퐇 of the sample remaining in the mixing chamber 24 for 2-fold dilution. Therefore, 5 占 퐇 of the buffer solution contained in the first metering chamber 31 To the mixing chamber (24). The first metering valve 56 is opened in a state in which the second metering valve 57 is closed. Thus, only 5 mu l of the buffer solution contained in the first metering chamber 31 flows into the mixing chamber 24 through the supply flow path 35. [ Thus, in the mixing chamber 24, 5 mu l of the sample is mixed with 5 mu l of the buffer solution to produce a diluted solution twice as much as the original sample concentration (see 3 in Fig. 5A)

After generating the sample dilution diluted twice in the mixing chamber 24, the dilution diluted twice in succession can be discharged from the mixing chamber 24 and stored in the storage chamber by repeating the procedure described above.

That is, in the mixing chamber 24, the diluting liquid is metered and discharged by a set amount, and only the metered sample remains in the mixing chamber 24. The diluting liquid discharged from the mixing chamber 24 into the first volume section 53 of the metering channel 51 is transferred along the discharge channel 61 and stored in the second reservoir chamber 63. The first storage valve 67 is closed and the second storage valve 68 is opened so that the diluting liquid is supplied to the second reservoir chamber 63 in the state in which the second shutoff valve 72 of the discharge passage 61 is closed, Lt; / RTI >

Then, the buffer solution is metered again in the first metering chamber 31, and then the metered buffer solution is supplied to the diluting solution remaining in the mixing chamber 24 and is re-diluted. The diluted diluted liquid is again metered through the metering channel 51 and stored in the third reservoir chamber 64 of the discharge channel 61. The diluting liquid is repeatedly metered and remained in the mixing chamber 24, and the buffer liquid is metered and supplied and mixed, whereby the diluting liquid can be continuously generated and stored at a constant magnification. In this embodiment, it is possible to successively produce a dilution liquid having a dilution ratio of 1, 2, 4, 8, and 16 times through the repetitive process. Diluent diluted at a certain ratio is sequentially stored in each storage chamber formed along the discharge passage 61.

FIG. 5 (B) shows a process of continuously diluting the sample at a dilution ratio of 10 times magnification.

By varying the operation of the valves, the samples can be serially diluted with dilution ratios of different magnifications through the same device. When the sample and the buffer solution are prepared in the body 10, the body 10 is rotated to apply centrifugal force to the fluid, and the valve is controlled and driven.

For the 10-fold dilution, the sample is weighed in the mixing chamber 24, and 9 μl of the sample is discharged, leaving only 1 μl of the remaining sample. The discharge valve 52 provided on the exit side of the mixing chamber 24 and the first metering valve 56 and the second metering valve 57 provided in the metering channel 51 are opened and the third metering valve 58 is closed do. Thus, the fluid in the mixing chamber 24 is filled in the first volume section 53, the second volume section and the third volume section 55.

Since the volume of the first volume section 53, the volume volume of the second volume section 54 and the volume volume of the third volume section 55 are preset to 5 μl, 3 μl and 1 μl respectively, the volume of the sample contained in the mixing chamber 24 9 mu l of the fluid is metered and discharged to the first volume section 53 to the third volume section 55 and only 1 mu l of the fluid remains in the mixing chamber 24. [

In this way, the fluid is transferred to the first volume section 53 to the third volume section 55 of the metering channel 51, so that only a fixed amount of fluid measured in the mixing chamber 24 can be left. (See 1 in Fig. 5 (B)).

After the sample is metered and remained in the mixing chamber 24, the sample discharged into the metering channel 51 is transferred to the first storage chamber 62 and stored.

The buffer solution is metered in a state in which a predetermined amount of the sample remains in the mixing chamber 24, and is transferred to the mixing chamber 24 to dilute the sample. When the open / close valve 34 provided on the outflow side of the buffer solution supply part 22 is opened, the buffer solution flows into each buffer solution metering chamber through the common flow path. After the buffer liquid metering chambers are filled, excess buffer liquid flows out to the waste chamber 41 along the waste flow path 40 connected to the common flow path, and is removed (refer to 2 in FIG. 5 (B)).

The buffer solution metered in the buffer solution metering chamber accommodating the buffer solution of the corresponding capacity in accordance with the dilution ratio is supplied to the mixing chamber 24 to dilute the sample with the desired dilution ratio.

In this embodiment, for the 10-fold dilution, 9 [mu] l of the buffer solution should be mixed with 1 [mu] l of the sample remaining in the mixing chamber 24. [ 3 μl of the buffer solution contained in the second metering chamber 32 as well as 5 μl of the buffer solution contained in the first metering chamber 31 and 1 μl of the buffer solution contained in the third metering chamber 33 The buffer solution is transferred to the mixing chamber 24. The first metering valve 56, the second metering valve 57 and the third metering valve 58 are all opened and the buffer solution contained in each metering chamber is supplied to the mixing chamber 24 ). Therefore, in the mixing chamber 24, 9 占 퐇 of buffer solution is mixed with 1 占 퐇 of the sample to produce a diluted solution which is diluted 10 times with respect to the initial sample concentration (see 3 in Fig. 5 (A)).

In this manner, the sample can be diluted with a desired dilution ratio by changing the operation of the valve provided in the flow path.

After the sample dilution diluted by 10 times in the mixing chamber 24 is produced, the dilution diluted by 10 times in succession can be discharged from the mixing chamber 24 and stored in the storage chamber by repeating the procedure described above.

That is, in the mixing chamber 24, the diluting liquid is metered and discharged by a set amount, and only the metered sample remains in the mixing chamber 24. The diluting liquid discharged from the mixing chamber 24 into the metering channel 51 is transported along the discharge channel 61 and is stored in the second reservoir chamber 63.

Then, the buffer solution is metered again in the first metering chamber 31, and then the metered buffer solution is supplied to the diluting solution remaining in the mixing chamber 24 and is re-diluted. The diluted diluted liquid is again metered through the metering channel 51 and stored in the third reservoir chamber 64 of the discharge channel 61. The diluting solution is repeatedly metered and remained in the mixing chamber 24, and the buffer solution is metered and supplied and mixed so that the diluting solution can be continuously generated and stored at a predetermined ratio. In this embodiment, a dilution liquid having a dilution ratio of 1x, 10x, 100x, 1000x, and 10000x can be continuously produced through the repetitive process. Diluent diluted at a certain ratio is sequentially stored in each storage chamber formed along the discharge passage 61.

By varying the valve operation in the same procedure described above, the sample can be diluted with dilution ratios of various magnifications in addition to dilution of 2 times or 10 times. For example, 2 占 퐇 of the sample may be left in the mixing chamber 24 and mixed with 8 占 퐇 of the metered buffer solution, or 5 占 퐇 of the buffer solution may be mixed with 1 占 퐇 of the sample, Samples can be serially diluted with a dilution ratio.

Further, a dilution liquid can be produced by varying the dilution ratio at an arbitrary magnification in addition to the dilution ratio of the fixed magnification through valve operation.

For example, a sample is continuously diluted at a dilution ratio of 5 times, 10 times, 2 times, and 5 times to continuously prepare a dilution liquid at a dilution ratio of 1, 5, 50, 100, and 500 times, Lt; / RTI >

The process of continuously diluting the diluent at the arbitrary dilution factor will be described with reference to FIG.

When the sample and the buffer solution are prepared in the body 10, the body 10 is rotated to apply centrifugal force to the fluid, and the valve is controlled and driven.

First, in order to prepare a 5-fold dilution, 8 쨉 l of sample is discharged from the sample in the mixing chamber 24, and only 2 쨉 l of the sample is left in the mixing chamber 24.

The discharge valve 52 provided on the outgoing side of the mixing chamber 24 and the first metering valve 56 provided on the metering channel 51 are opened and the second metering valve 57 is closed. The fluid in the mixing chamber 24 is discharged to the outlet side of the mixing chamber 24 by centrifugal force and flows along the metering channel 51. The fluid discharged from the mixing chamber 24 is supplied to the second metering valve 57 of the metering channel 51 since the second metering valve 57 of the metering channel 51 is closed and the metering channel 51 is closed, So that the first volume section 53 and the second volume section 54 are filled.

Since the volume of the first volume section 53 and the volume volume of the second volume section 54 are preset to be 5 μl and 3 μl, 8 μl of the fluid contained in the mixing chamber 24 is metered in the first volume section 53 and the second volume section 54 and only 2 l of fluid can be left in the mixing chamber 24.

As described above, the fluid is transferred to the first volume section 53 and the second volume section 54 of the metering channel 51, so that only a fixed amount of fluid metered into the mixing chamber 24 can be left.

In addition to the sample weighing, the buffer solution supply unit 22 transfers the buffer solution to each buffer solution weighing chamber and weighs it. 5 mu l of buffer solution is accommodated in the first metering chamber 31 and 3 mu l of buffer solution is accommodated in the second metering chamber 32 and 1 mu l of buffer solution is accommodated in the third metering chamber 33 .

The sample filled in the metering channel 51 is discharged from the metering channel 51 and flows into the first storage chamber 62 of the discharge channel 61 to be stored do. In the first storage chamber 62, a sample of the initial concentration is stored.

With the 2 의 sample left in the mixing chamber 24, the buffer solution metered to the dilution ratio is supplied to the mixing chamber 24 to dilute the sample. For the 5-fold dilution, 8 [mu] l of the buffer solution should be mixed with 2 [mu] l of the sample remaining in the mixing chamber 24, so that 5 [mu] l of the buffer solution contained in the first metering chamber 31, 32) is supplied to the mixing chamber (24). The first metering valve 56 and the second metering valve are opened in a state in which the third metering valve 58 is closed. Thus, only the buffer solution contained in the first metering chamber 31 and the second metering chamber 32 flows into the mixing chamber 24 through the supply flow path 35. Thus, in the mixing chamber 24, 8 占 퐇 of the buffer solution is mixed with 2 占 퐇 of the sample, and a diluted solution which is 5 times diluted from the initial sample concentration is produced. (See (C) in Fig. 6)

In order to continuously dilute the dilution liquid 10 times, 9 l of the dilution liquid is metered out from the dilution liquid in the mixing chamber 24, and only 1 l of the dilution liquid is left in the mixing chamber 24. The 10-fold dilution procedure is the same as that described with reference to FIG. 5 (B), and a detailed description thereof will be omitted. 9 L of the 5-fold diluted solution metered and discharged in the mixing chamber 24 is discharged to the second storage chamber 63 through the discharge passage 61 and stored. Thus, the dilution liquid diluted five times with respect to the initial sample concentration is stored in the second reservoir chamber 63. (Refer to (D) to (D) in FIG. 6)

In order to continuously dilute the diluting liquid twice, 5 l of the diluting liquid is metered out from the diluting liquid in the mixing chamber 24, and only 5 l of the diluting liquid is left in the mixing chamber 24. The double dilution process is the same as that described with reference to FIG. 5 (A), and therefore, detailed description thereof will be omitted. 5 mu l of the 10-fold diluted solution metered and discharged in the mixing chamber 24 is discharged to the third storage chamber 64 through the discharge flow path 61 and stored. Thus, the dilution liquid diluted 50 times with respect to the initial sample concentration is stored in the third storage chamber 64 (refer to (E) through (F) of FIG. 6).

In order to continuously dilute the dilution liquid 5 times, 8 占 퐇 of the diluting liquid is metered out from the diluting liquid in the mixing chamber 24, and only 2 占 퐇 of the diluting liquid is left in the mixing chamber 24. The 5-fold dilution procedure is the same as that described above, and therefore, detailed description thereof will be omitted. 8 mu l of the double diluted solution metered and discharged in the mixing chamber 24 is discharged to the fourth storage chamber 65 through the discharge flow path 61 and stored. Thus, the dilution liquid diluted 100 times with respect to the initial sample concentration is stored in the fourth storage chamber 65. (See (S) through (A) in FIG. 6)

Finally, the diluent contained in the mixing chamber 24 is discharged and stored in the fifth storage chamber 66. In the mixing chamber 24, 8 [mu] l of buffer solution is mixed with 2 [mu] l of dilution solution for 5 times dilution, and the resultant solution is transferred along the metering channel 51 and the discharge channel 61 to the fifth storage chamber 66 . Thus, the dilution liquid diluted to 500 times the initial sample concentration is stored in the fourth storage chamber 65 (see FIG. 6)

The dilution liquid can be continuously prepared by changing it to a randomly set magnification such as 1, 5, 50, 100, or 500 times rather than a fixed magnification through the above-described repetitive process.

Thus, the present apparatus can continuously produce a dilution liquid at an arbitrary ratio through a single apparatus by simple valve operation, sequentially store the prepared dilution liquid in each of the storage chambers according to the dilution ratio, It can be easily performed.

[Experimental Example]

The graph of FIG. 7 shows the results of microfluid dilution using the diluting device according to the present embodiment.

The experiment was carried out by measuring the result of four successive dilutions of the fluorescence solution at a magnification of 2 with the dilution apparatus of this embodiment and the results of three consecutive dilutions at a magnification of 10.

7 (a) shows the result of continuous dilution at 2-fold magnification using the log value, and (b) shows the result of continuous dilution at 10-fold magnification using the log value.

As shown in the graph of FIG. 7, it can be confirmed that the logarithm of each magnification is used to show excellent linearity in the analysis. Thus, it can be seen that continuous dilution is properly performed through the apparatus. In addition, it was confirmed that the continuous dilution process for four consecutive dilutions can be performed mechanically in a short time with a time of about 10 minutes.

While the present invention has been described in connection with what is presently considered to be practical exemplary embodiments, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, Of course.

10: Body 20:
22: buffer solution supply unit 24: mixing chamber
30: buffer solution metering section 31: first metering chamber
32: second metering chamber 33: third metering chamber
34: opening / closing valve 35:
36: first supply valve 37: second supply valve
38: third supply valve 39: common flow path
40: waste channel 41: waste chamber
42: disposal valve 43: air passage
44: air hole 50: sample metering section
51: metering channel 52: discharge valve
53: first volume section 54: second volume section
55: third volume section 56: first metering valve
57: Second metering valve 58: Third metering valve
60: storage part 61: discharge channel
62, 63, 64, 65,
67, 68, 69, 70: Storage valves 71, 72, 73, 74:

Claims (19)

A body having a flow path through which the fluid is delivered and a valve for selectively opening and closing the flow path to control the flow of the fluid, a dilution part formed along the flow path in the body for diluting the sample and a dilution part connected to the dilution part, And a storage unit,
The dilution unit may include a buffer solution supply unit for supplying the buffer solution, a mixing chamber connected to the buffer solution supply unit to dilute the buffer solution mixed with the sample, and a mixing chamber connected to the mixing chamber out side, And a sample metering section for allowing the metered solution to remain,
The sample metering section includes a metering channel connected to the outflow side of the mixing chamber for discharging the solution, a discharge valve for opening and closing the outflow side of the mixing chamber, and a metering channel provided at intervals along the metering channel to open and close the metering channel, And at least one metering valve for partitioning into at least one volume section. The method according to claim 1,
And a sample supply unit connected to the mixing chamber and configured to transfer the sample to the mixing chamber. The method according to claim 1,
And a buffer liquid metering unit connected between the buffer solution supply unit and the mixing chamber for metering and supplying the buffer solution. The method of claim 3,
Wherein the buffer solution metering section comprises at least one or more buffer solution metering chambers disposed on the outward side of the buffer solution supply section for metering and storing a buffer solution, an on / off valve provided on the out side of the buffer solution supply section, And at least one supply valve provided in the supply passage formed between the supply passage and the supply passage and opening / closing the supply passage. 5. The method of claim 4,
The buffer solution metering unit further includes a waste flow channel connected to the buffer solution metering chamber to discharge excess solution exceeding the volume of the metering chamber and a waste chamber connected to the waste flow channel to receive excess solution discarded through the waste flow channel Lt; / RTI > 5. The method of claim 4,
Wherein said buffer solution metering section has the same volume as each of said buffer solution metering chambers or at least one of said metering chambers has a different volume. delete The method according to claim 1,
Wherein the metering channels have the same volume of each volume section, or at least one volume section of the metering channel has a different volume. The method according to claim 1,
Wherein the body has a rotation center to apply a centrifugal force to the fluid to transfer the fluid along the flow path. The method according to claim 1,
And an air hole formed in the body and communicating with the flow path and the outside. The method according to claim 1,
Wherein the reservoir includes at least one storage chamber connected to the diluter outlet for receiving the diluent transferred through the diluter, and a storage valve for opening and closing each storage chamber inlet. 12. The method of claim 11,
Wherein the storage unit is sequentially connected to a plurality of storage chambers along a discharge channel connected to the sample metering unit and the shut-off valve is installed in the discharge channel between the storage chambers. 12. The method of claim 11,
Wherein the buffer solution supply unit, the buffer solution metering unit, the mixing chamber, the sample metering unit, and the storage unit are sequentially arranged outward from the center of the body and connected in series. A sample and a buffer solution are supplied to the main body,
The sample accommodated in the mixing chamber of the main body is discharged in a fixed amount, and the metered sample is left in the mixing chamber,
The buffer solution is transferred to the mixing chamber and mixed with the residual sample to dilute,
Discharging and storing the diluent in the mixing chamber,
Transferring the buffer solution to a plurality of different volume metering chambers prior to transferring the buffer solution to the mixing chamber,
Further comprising transferring a buffer solution of at least one metering chamber to the mixing chamber according to a dilution ratio. delete 15. The method of claim 14,
And in the buffer liquid metering step, removing excess quantities of solution in each metering chamber. 17. The method according to claim 14 or 16,
Upon dispensing the diluent in the mixing chamber,
The diluted solution in the mixing chamber is discharged in a fixed amount, the diluted solution is left in the mixing chamber,
The buffer solution is transferred to the mixing chamber, mixed with a diluting solution and re-diluted,
Diluting the diluted solution in the mixing chamber, storing the diluted solution in the mixing chamber, and continuously diluting the diluted solution by repeating the process of re-diluting, discharging, and storing the remaining diluted solution. 18. The method of claim 17,
Wherein the continuous dilution is performed by repeating dilution with a sample of the same volume and a buffer solution of the same volume to continuously dilute the same in the same ratio. 18. The method of claim 17,
Wherein the continuous dilution is performed by repeating dilution with different volumes of sample and different volumes of buffer solution to continuously dilute the sample in an arbitrary ratio.

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