阅读说明：本技术 生物化学用盒和生物化学分析装置 (Cartridge for biochemistry and biochemical analyzer ) 是由 藤冈满 足立作一郎 于 2018-10-19 设计创作，主要内容包括：为了能够在距EWOD电极具有规定距离的位置抽取生物体试样,本发明的生物化学用盒的特征在于,具备：液滴流路,其中,输送包含生物体试样的液滴即试样液滴的多个EWOD电极沿着输送所述试样液滴的方向排列；以及试样抽取部,其位于距所述液滴流路的末端的EWOD电极具有规定的距离、且可抽取所述试样液滴中的所述生物体试样的位置；在抽取所述生物体试样时,所述试样抽取部位于比具有所述末端的EWOD电极的所述液滴流路低的位置,所述液滴流路与所述试样抽取部之间平滑地相连。(In order to enable the extraction of a biological sample at a position spaced apart from an EWOD electrode by a predetermined distance, a biochemical cartridge according to the present invention includes: a droplet channel in which a plurality of EWOD electrodes that transport sample droplets, which are droplets containing biological samples, are arranged along a direction in which the sample droplets are transported; and a sample extraction unit that is located at a position that is a predetermined distance from the EWOD electrode at the end of the droplet channel and from which the biological sample in the sample droplet can be extracted; when the biological sample is extracted, the sample extraction unit is located at a position lower than the droplet channel of the EWOD electrode having the tip, and the droplet channel and the sample extraction unit are smoothly connected to each other.)

1. A biochemical cartridge is provided with:

a droplet channel in which a plurality of EWOD electrodes that transport sample droplets, which are droplets containing biological samples, are arranged along a direction in which the sample droplets are transported; and

a sample extraction unit that is provided at a position that is a predetermined distance from the EWOD electrode at the end of the droplet channel and from which the biological sample in the sample droplet can be extracted;

when the biological sample is extracted, the sample extraction unit is located at a position lower than the droplet channel of the EWOD electrode having the tip, and the droplet channel and the sample extraction unit are smoothly connected to each other.

2. The biochemical cartridge according to claim 1, wherein the droplet channel and the sample collection unit are included in a transport device, and the transport device is kept horizontal when not subjected to a load and is inclined when subjected to a load when collecting the biological sample.

3. The biochemical cartridge according to claim 2, wherein the transport device is inclined by a load caused by a capillary array for extracting the biological sample.

4. The biochemical cartridge according to claim 1, wherein a track is provided between the droplet channel and the sample collection unit.

5. The biochemical cartridge according to claim 1, wherein the sample drawing section has a droplet retaining section for retaining the sample droplet.

6. The biochemical cartridge according to claim 5, wherein the droplet holding part slides when the sample droplet is extracted.

7. The cartridge for biochemistry of claim 1, wherein the droplet channel and the sample drawing section are included in a transport device, and the transport device is inclined with respect to a horizontal plane.

8. The cartridge for biochemistry of claim 1, wherein the droplet channel is kept horizontal, and the droplet channel and the sample collection unit are inclined with respect to a horizontal plane.

9. The biochemical cartridge according to claim 1, wherein a distance from the distal EWOD electrode to the sample extraction portion is a distance that maintains electrical insulation between the sample extraction portion and the distal EWOD electrode with respect to an applied voltage for extracting the biological sample.

10. A biochemical analyzer for extracting and analyzing a biological sample, comprising the biochemical cartridge according to claim 1.

Technical Field

The present invention relates to a biochemical cartridge for synthesizing and analyzing a biological sample extracted by a biochemical reaction as necessary, and a biochemical analyzer including the biochemical cartridge.

Background

Genomic analysis such as nucleotide sequence analysis and polymorphism analysis is very important in the fields of biological research, medical fields such as gene therapy, diagnosis and development of molecular targeted drugs, and forensic fields such as DNA identification. In the genome analysis, 1) a step of extracting nucleic acid from a sample, 2) a step of amplifying the extracted nucleic acid and labeling the amplified nucleic acid, and 3) a step of electrophoresis for reading the nucleotide sequence of the nucleic acid are performed. 2) In the step (2), the nucleic acid mixed with the reagent is maintained at a predetermined temperature, whereby the primer anneals to the target nucleic acid to amplify the nucleic acid.

Patent document 1 discloses a method of using electrowetting on dielectric (EWOD; electro Wetting On direct). That is, patent document 1 discloses that droplets of nucleic acids and reagents are transported in a droplet microactuator using EWOD, nucleic acids are amplified, and then analyzed downstream by electrophoresis.

Documents of the prior art

Patent document

Patent document 1: japanese Kokai publication Hei-2009-534653

Disclosure of Invention

Problems to be solved by the invention

However, patent document 1 does not disclose a specific method of supplying amplified nucleic acids to an electrophoresis apparatus such as a capillary sequencer. While EWOD transports droplets with an applied voltage of several tens of V, it requires an applied voltage of several kV in order to extract a biological sample in the droplets, for example, amplified nucleic acid, into a capillary array of a capillary sequencer or the like, and thus EWOD electrodes are broken and cannot be reused for transporting droplets.

Accordingly, an object of the present invention is to provide a biochemical cartridge capable of extracting a biological sample at a position spaced apart from an EWOD electrode by a predetermined distance, and a biochemical analyzer including the biochemical cartridge.

Means for solving the problems

In order to achieve the above object, a biochemical cartridge according to the present invention includes: a droplet channel in which a plurality of EWOD electrodes that transport sample droplets, which are droplets containing biological samples, are arranged along a direction in which the sample droplets are transported; and a sample extraction unit that is located at a position that is a predetermined distance from the EWOD electrode at the end of the droplet channel and from which the biological sample in the sample droplet can be extracted; when the biological sample is extracted, the sample extraction unit is located at a position lower than the droplet channel of the EWOD electrode having the tip, and the droplet channel and the sample extraction unit are smoothly connected to each other.

The present invention is a biochemical analyzer for extracting and analyzing a biological sample, including the biochemical cartridge.

Effects of the invention

According to the present invention, it is possible to provide a biochemical cartridge capable of extracting a biological sample at a position spaced apart from an EWOD electrode by a predetermined distance, and a biochemical analyzer including the biochemical cartridge.

Drawings

FIG. 1 is a diagram showing the overall configuration of a biochemical analyzer according to example 1.

FIG. 2 is a perspective view illustrating the biochemical cartridge according to example 1.

FIG. 3 is a plan view showing the sample flow path, the reagent flow path and the sample collection unit in the biochemical cassette according to example 1.

FIG. 4 is a cross-sectional view illustrating an EWOD (electro Wetting On direct) device of example 1.

FIG. 5 is a sectional view illustrating a sample collection unit according to example 1.

FIG. 6 is a flowchart for explaining the operation of example 1.

FIG. 7 is a sectional view for explaining the operation of example 1.

FIG. 8 is a sectional view for explaining the operation of example 1.

FIG. 9 is a sectional view for explaining the operation of example 1.

FIG. 10 is a sectional view showing a sample collection portion according to example 2.

FIG. 11 is a flowchart for explaining the operation of example 2.

FIG. 12 is a sectional view showing a sample collection portion according to example 3.

FIG. 13 is a flowchart for explaining the operation of example 3.

FIG. 14 is a sectional view showing a sample collection portion according to example 4.

Detailed Description

Hereinafter, an embodiment of the biochemical analyzer according to the present invention will be described with reference to the drawings. Note that XYZ coordinate systems are attached to the respective drawings to show the orientation of the respective drawings.

Example 1

Fig. 1 shows the overall structure of a biochemical analysis apparatus. In this example, a biochemical analyzer is exemplified in which nucleic acids extracted from a sample are amplified and labeled, and then the base sequence of the nucleic acids is read. The nucleic acid mixed with the reagent is maintained at a predetermined temperature for amplifying the nucleic acid, or the amplified nucleic acid is supplied to a capillary called a capillary for electrophoresis.

The apparatus main body 101 and the control computer 125 are connected by a communication cable. The control computer 125 receives an input from an operator, controls each function of the biochemical analyzer, transmits and receives data detected by the apparatus main body 101, and displays the transmitted and received data. The apparatus main body 101 includes a capillary array 114, a pump mechanism 103, a thermostatic bath 115, a carrier 122, a high-voltage power supply 104, a light source 111, and an optical detector 112. Hereinafter, each part will be explained.

The capillary array 114 is a replacement member including one or more (for example, 2 to 96) capillaries 102, and includes a loading head 124, a detection section 113, and a capillary head 129. A loading head 124 for supplying a sample into the capillary 102 is provided at one end of the capillary array 114, and a cathode end 126 for applying a negative voltage is formed. The other end of the capillary array 114 is connected to the gel block 106 by bundling a plurality of capillaries 102 into one piece with a capillary head 129 and using a pressure-resistant airtight structure. A detection unit 113 for irradiating laser light is provided between the loading head 124 and the capillary head 129.

The capillary 102 is a glass tube having an inner diameter of several tens to several hundreds of μm and an outer diameter of several hundreds of μm. In order to increase the strength of the capillary 102, the surface thereof is covered with a polyimide film. However, the polyimide film is removed at the detection section 113 irradiated with the laser beam and its vicinity. The capillary 102 is filled with a separation medium for separating DNA molecules in a sample. The separation medium is, for example, a polyacrylamide-based separation gel.

The pump mechanism 103 is constituted by a syringe 105 and a mechanism system for pressurizing the syringe 105. The gel block 106 is a connection part for connecting the syringe 105, the capillary array 114, the anode buffer container 108, and the separation medium container 107. By closing the electric valve 110 and pressing the syringe 105, the separation medium in the syringe 105 is injected into the capillary 102.

The thermostatic bath 115 has a heater 117 and a fan 116 for controlling the temperature of the capillary array 114, and is covered with an insulating material in order to keep the temperature inside the thermostatic bath 115 constant. By controlling the temperature within the thermostatic bath 115, the temperature of most of the capillary array 114 is maintained at a constant temperature, for example, 60 ℃.

The conveyor 122 includes 3 electric motors and linear actuators, and is movable in 3 axial directions of up and down, right and left, and front and back. At least 1 or more containers are mounted on the movable table 123 of the conveyor 122. The conveyor 122 conveys the buffer container 118, the cleaning container 119, the waste liquid container 120, and the biochemical cartridge 121 on the moving stage 123 to the cathode end 126 of the capillary 102. The running buffer enters the buffer container 118. The cleaning vessel 119 is used to clean the capillary 102. The separation medium in capillary 102 is discharged to waste reservoir 120. A biological sample, for example, a nucleic acid and a reagent are put into the biochemical cartridge 121, and the nucleic acid amplified in the biochemical cartridge 121 is extracted from the cathode end 126 of the capillary 102 into the capillary array 114. The biochemical cartridge 121 will be described later using fig. 2 to 5.

The high voltage power supply 104 is connected to the anode electrode 109 and the loading head 124 in the anode buffer container 108, and applies a high voltage to the separation medium in the capillary 102.

The light source 111 irradiates the detection unit 113 with laser light as coherent light as excitation light. The optical detector 112 optically detects fluorescence emitted from the sample in the detection section 113. The detected optical data 128 is transmitted to the control computer 125 via the control board 127.

The biochemical cartridge 121 will be described with reference to fig. 2. Fig. 2 is a perspective view of the biochemical cartridge 121. The biochemical cartridge 121 is provided with 1 or more, for example, 4 channels for amplified nucleic acids, and the cathode end 126 of the capillary 102 is inserted into each channel. In fig. 2, the longitudinal direction of the flow channels is indicated by the X direction, the direction in which the flow channels are arranged is indicated by the Y direction, and the direction in which the cathode end 126 is inserted is indicated by the Z direction.

The structure in the biochemical cartridge 121 will be described with reference to fig. 3. Fig. 3 is a plan view of the inside of the biochemical cartridge 121. The biochemical cartridge 121 of fig. 3 is provided with a sample well 301, a reagent well 302, a sample channel 303, and a reagent channel 304. A plurality of, for example, 4 sample wells 301 are provided, and 1. mu.L of nucleic acid to be amplified is placed in each sample well 301. Alternatively, 10. mu.L of nucleic acid may be put into the sample well 301 and 1. mu.L of nucleic acid may be separated from 10. mu.L of nucleic acid and used. The reagent wells 302 are provided with 1 or more, for example, 5, and each reagent well 302 is filled with a reagent for nucleic acid amplification, for example, a primer, dNTP, buffer, water, enzyme, modifier, molecular weight standard DNA, or the like.

The sample channel 303 is connected to each sample cell 301, and transports a droplet containing nucleic acid. In this example, the direction in which the droplets containing the nucleic acid were transported was the X direction. When the EWOD (electro Wetting On direct) technique is used for transporting droplets, the sample channel 303 serves as a transport device having the EWOD electrode 300 for transporting droplets. EWOD refers to the following technique: a voltage is applied between the droplets disposed on the water-repellent film as the water-repellent film and an EWOD electrode as an electrode disposed below the water-repellent film, and the surface tension of the droplets is controlled, whereby the droplets are transported on the water-repellent film.

An example of a transport apparatus using EWOD will be described with reference to fig. 4. Fig. 4 is an XZ sectional view of the conveying apparatus. The transport apparatus has an upper plate 401, an upper electrode 402, an upper water repellent film 403, a lower water repellent film 405, an insulating film 406, an EWOD electrode 300, and a lower plate 407. The upper plate 401 and the lower plate 407 are arranged in parallel, an upper electrode 402 and an upper water repellent film 403 are provided on the lower surface of the upper plate 401, and a plurality of EWOD electrodes 300, an insulating film 406, and a lower water repellent film 405 are provided on the upper surface of the lower plate 407. Further, if a plurality of EWOD electrodes 300 are disposed on at least one of the upper plate 401 and the lower plate 407, the droplet 400 can be transported.

A plurality of EWOD electrodes 300 are aligned along a direction in which the droplets 400 are transported. In addition, the EWOD electrodes 300 are covered with an insulating film 406 having a thickness of, for example, several hundreds of μm so that a voltage can be applied individually to each EWOD electrode 300. The space between the upper and lower water repellant films 403, 405 is preferably filled with fluid 404 that does not mix with the transported droplets 400. Further, even without the fluid 404, the droplet 400 can be transported.

In such a transport apparatus, when a voltage of several tens of V is applied to the EWOD electrode 300 located in the vicinity of the droplet 400, the surface tension of the droplet 400 on the EWOD electrode 300 side to which the voltage is applied changes, and an internal pressure is generated in the droplet 400. The generated internal pressure drives the droplet 400 in the direction of the arrow in fig. 4, and thus the droplet 400 is conveyed. That is, the droplet 400 is transported to the EWOD electrode 300 side to which the voltage is applied.

Returning to the description of fig. 3. The reagent flow path 304 is connected to each reagent tank 302 and transports a droplet of the reagent. In this embodiment, the direction in which the reagent droplets are transported is the Y direction. When EWOD is used for transporting reagent droplets, the reagent channel 304 has a plurality of EWOD electrodes 300, as in the case of the sample channel 303. The reagent channel 304 intersects the sample channel 303, and a droplet of the reagent and a droplet of the nucleic acid are mixed at the intersection of the two. The angle at which the reagent channel 304 and the sample channel 303 intersect is not limited to 90 degrees as shown in fig. 3.

Since a voltage can be applied to the EWOD electrodes 300 of the sample channel 303 and the reagent channel 304 individually, 2 or more droplets can be simultaneously transferred. The direction in which the droplets are transported is not limited to one direction, and the droplets may be reciprocated. For example, the droplet may be reciprocated between the intersection of the sample channel 303 and the reagent channel 304 and a point adjacent to the intersection to promote the mixing of the nucleic acid and the reagent.

A temperature control region 305 is provided in the middle of the sample channel 303. The temperature control region 305 is a region maintained at 1 or more predetermined temperatures, for example, a region maintained at 60 ℃ and a region maintained at 95 ℃. The droplets mixed with the nucleic acid and the reagent are transported to the temperature control region 305, and the nucleic acid is amplified by, for example, PCR (Polymerase Chain Reaction) or cycle sequencing Reaction. The droplets may be reciprocated between regions maintained at different temperatures, for example, between a region at 60 ℃ and a region at 95 ℃. The droplet after the nucleic acid amplification is labeled to become a sample droplet. The sample channel 303 for transporting a sample droplet is also referred to as a droplet channel.

A sample collection unit 306 is provided at the tip of the sample channel 303. In the sample extracting section 306, the nucleic acid in the sample droplet is extracted to the capillary array 114. That is, the cathode end 126 of the capillary 102 is inserted at the position of the sample collection portion 306. The sample extraction unit 306 is located at a predetermined distance from the EWOD electrode 300A at the end of the sample channel 303, and is located at a distance that can electrically insulate the EWOD electrode 300A at the end when a voltage of several kV is applied to the loading head 124 of the capillary array 114, for example.

The transport facility 500 in the biochemical cassette 121 will be described with reference to fig. 5. Fig. 5(a) is an XZ sectional view of the transport apparatus 500 in the biochemical cartridge 121. Fig. 5(b) is a view of the section a-a in fig. 5 (a). The transport apparatus 500 includes an upper plate 401, an upper electrode 402, an upper water repellent film 403, a lower water repellent film 405, an insulating film 406, an EWOD electrode 300, a lower plate 407, and an opening 501. The upper plate 401, the upper electrode 402, the upper water repellent film 403, the lower water repellent film 405, the insulating film 406, the EWOD electrode 300, and the lower plate 407 have the same structure as that shown in fig. 4, and the sample droplet 502 is transported by this structure. Opening 501 is provided at the position of sample collection unit 306, and cathode end 126 of capillary 102 is inserted. The sample extracting unit 306 is indicated by an ellipse of a broken line in the figure.

A fulcrum part 503 and a supporting part 504 are provided between the biochemical cartridge 121 and the transport apparatus 500. The fulcrum portion 503 supports the transport apparatus 500 rotatably about the Y axis at a position on the + X direction side of the center of gravity of the transport apparatus 500. The support portion 504 supports the conveyance device 500 so as to be horizontal together with the fulcrum portion 503, and is disposed on the-X direction side of the fulcrum portion 503. That is, when the conveyance device 500 is not subjected to a load, the conveyance device 500 is kept horizontal, and when a load in the + Z direction is applied to a position on the + X direction side of the fulcrum portion 503, the conveyance device 500 is inclined with respect to the horizontal plane. By tilting the transport apparatus 500, the sample extraction section 306 is at a position lower than the droplet flow path including the distal EWOD electrode 300A. The droplet channel and the sample collection unit 306 are smoothly connected to each other by the lower water repellent film 405.

Further, a rut 505 shown in fig. 5(b) may be provided on the lower water repellent film 405 between the droplet channel and the sample collection unit 306. The switch 505 is provided along the X direction, i.e., along the droplet flow path, from the position where the EWOD electrodes 300 are arranged. By providing the track 505 between the droplet channel and the sample extraction unit 306, the sample droplet 502 can be prevented from being deviated in the Y direction.

The fulcrum 503 and the supporting portion 504 may be provided between the movable stage 123 and the biochemical cartridge 121, instead of between the biochemical cartridge 121 and the transport apparatus 500. In this case, the biochemical cartridge 121 is inclined or kept horizontal with respect to the horizontal plane depending on the presence or absence of the load.

The operation of the present embodiment will be described with reference to fig. 6.

(S601)

By controlling the applied voltage to the EWOD electrode 300 of the delivery apparatus 500, the sample droplet 502 is delivered to the location of the end EWOD electrode 300A. Fig. 5(a) shows a case where the sample droplet 502 is transported to the position of the EWOD electrode 300A at the end.

(S602)

The capillary array 114 moves in the + Z direction, passes through the opening 501, and then comes into contact with the lower water repellent film 405. The transport device 500 receives a load in the + Z direction of the capillary array 114, and is inclined with respect to the horizontal plane with the contact portion with the fulcrum portion 503 as a rotation axis. Fig. 7 shows a state in which the conveying apparatus 500 is inclined. The transport device 500 is tilted to generate a driving force in the direction of the arrow in fig. 7 for the sample droplet 502, but the sample droplet 502 is held at the position of the EWOD electrode 300A at the end by applying a voltage to the EWOD electrode 300.

(S603)

The applied voltage to the EWOD electrode 300 is set to an open or grounded state. When the applied voltage is turned off or grounded, the force holding the sample droplet 502 at the position of the EWOD electrode 300A at the end disappears, and therefore the sample droplet 502 starts moving in the direction of the arrow in fig. 7. The sample droplet 502 moves along the upper surface of the lower water repellent film 405, and stops at the position of the cathode end 126 of the capillary 102 due to surface tension. Fig. 8 shows a case where sample droplet 502 stops at the position of cathode end 126 of capillary 102. Further, a voltage of about several V may be applied to the capillary 102 in order to make the sample droplet 502 more likely to remain at the position of the cathode end 126 of the capillary 102.

(S604)

A voltage of several kV is applied to the loading head 124. By applying a voltage to the loading head 124, a predetermined amount of nucleic acid in the sample droplet 502 stopped at the position of the cathode end 126 of the capillary 102 is drawn into the capillary array 114. When a predetermined amount of nucleic acid is extracted into the capillary array 114, the voltage applied to the loading head 124 is turned off.

(S605)

After the voltage application to the loading head 124 is turned off, the capillary array 114 moves in the-Z direction away from the lower water repellent film 405. The capillary array 114 is separated from the lower water repellent film 405, so that the load on the transport device 500 is eliminated, and the transport device 500 returns to the horizontal state. Fig. 9 shows a case where the capillary array 114 is separated from the lower water repellent film 405 and the transport facility 500 is returned to a horizontal state. The sample droplet 502 remaining after the nucleic acid is extracted into the capillary array 114 moves in the direction of the arrow in fig. 9 while the transport device 500 is tilted, and is discharged from the transport device 500.

According to the present embodiment described above, in the sample extracting section 306 provided at a position having a predetermined distance from the EWOD electrode 300A at the end of the transport apparatus 500, the nucleic acid in the sample droplet 502 can be extracted to the capillary array 114. Since the sample extraction unit 306 is located at a position where electrical insulation can be maintained even when a voltage necessary for extracting nucleic acid by the capillary array 114 is applied to the loading head 124, dielectric breakdown of the EWOD electrode can be prevented. That is, according to the present embodiment, the transport apparatus 500 including the EWOD electrode 300 can be used a plurality of times when extracting nucleic acid using the capillary array 114.

In this example, the case of treating nucleic acids, particularly DNA, has been described as an example of a biological sample. The biological sample to be treated in the present invention is not limited to DNA, and includes all biological substances such as RNA, protein, polysaccharide, and microorganism. In addition, a member other than the capillary 102 may be used for the extraction of the biological sample.

Example 2

In example 1, a case where the transport device 500 is tilted by the load of the capillary array 114 to move the sample droplet 502 to the sample extracting section 306 is described. Loads other than capillary array 114 may be used to tilt transport apparatus 500. Therefore, in the present embodiment, a case where the transport apparatus 500 is tilted by a load other than the capillary array 114 will be described.

This embodiment will be described with reference to fig. 10. Fig. 10 is an XZ sectional view of the conveyance device 500, as in fig. 5 (a). The transport apparatus 500 of the present embodiment has an upper plate 401, an upper electrode 402, an upper water repellent film 403, a lower water repellent film 405, an insulating film 406, an EWOD electrode 300, a lower plate 407, and an opening 501, and has a droplet retention section 1001, as in embodiment 1. Further, between the transport facility 500 and the biochemical cartridge 121, the supporting point part 503 and the elevating part 1000 are provided as in example 1. The supporting point 503 and the elevating unit 1000 may be provided between the biochemical cartridge 121 and the movable stage 123. The upper plate 401, the upper electrode 402, the upper water repellent film 403, the lower water repellent film 405, the insulating film 406, the EWOD electrode 300, the lower plate 407, the opening 501, and the fulcrum portion 503 are the same as those in embodiment 1, and therefore, description thereof is omitted. The droplet holding section 1001 is disposed in the vicinity of the opening 501, and is a member for holding the sample droplet 502 at the position of the sample extraction section 306. If necessary, the droplet holding portion 1001 may be slid in the Z direction or the Y direction.

The elevating unit 1000 is a driving device that extends and contracts in the-Z direction from the same height as the fulcrum unit 503, and is configured by, for example, a linear motor or the like, and is disposed on the-X direction side of the fulcrum unit 503. When the elevating unit 1000 and the fulcrum unit 503 are at the same height, the transport device 500 is kept horizontal, and when the elevating unit 1000 extends in the-Z direction, the transport device 500 is inclined with respect to the horizontal plane with the contact portion with the fulcrum unit 503 as a rotation axis.

The operation of the present embodiment will be described with reference to fig. 11.

(S1101)

By controlling the voltage applied to the EWOD electrode 300 of the transport apparatus 500, the sample droplet 502 is transported to the position of the end EWOD electrode 300A.

(S1102)

The lifting portion 1000 extends in the-Z direction. The conveyance device 500 is subjected to a load in the-Z direction of the lifting unit 1000, and is inclined with respect to the horizontal plane with the contact portion with the fulcrum portion 503 as a rotation axis. Fig. 10 shows a state in which the conveying apparatus 500 is inclined. The transport device 500 is tilted to generate a driving force in the direction of the arrow in fig. 10 on the sample droplet 502, but the EWOD electrode 300 is held at the position of the distal EWOD electrode 300A because a voltage is applied to the EWOD electrode 300.

(S1103)

The applied voltage to the EWOD electrode 300 is set to an open or grounded state. When the applied voltage is turned off or grounded, the force holding the sample droplet 502 at the position of the EWOD electrode 300A at the end disappears, and therefore the sample droplet 502 starts moving in the direction of the arrow in fig. 10. The sample droplet 502 moves along the upper surface of the lower water repellent film 405, and is retained at the position of the sample extraction unit 306 by the droplet retaining unit 1001.

(S1104)

The capillary array 114 moves in the + Z direction, and is inserted into the transport device 500 through the opening 501, and the cathode end 126 of the capillary 102 comes into contact with the sample droplet 502.

(S1105)

The nucleic acid in the sample droplet 502 is extracted to the capillary array 114 by applying a voltage of several kV to the loading head 124. After the nucleic acid in the sample droplet 502 is introduced into the capillary array 114 by a predetermined amount, the voltage applied to the loading head 124 is turned off.

(S1106)

The capillary array 114 moves in the-Z direction and the droplet holding portion 1001 slides in the Z direction or the Y direction, so that the sample droplet 502 remaining after the nucleic acid is extracted into the capillary array 114 moves on the inclined surface and is discharged from the transport device 500. By discharging the remaining sample droplet 502, mixing into the sample droplet 502 to be subsequently transported can be prevented. After the sample droplet 502 is discharged, the elevating portion 1000 contracts to the same height as the fulcrum portion 503, so that the transport apparatus 500 returns to the horizontal state.

According to the present embodiment described above, in the sample extracting section 306 provided at a position having a predetermined distance from the EWOD electrode 300A at the end of the transport apparatus 500, the nucleic acid in the sample droplet 502 can be extracted to the capillary array 114. Since the sample extraction unit 306 is located at a position where electrical insulation can be maintained even when a voltage necessary for extracting nucleic acid from the capillary array 114 is applied to the loading head 124, dielectric breakdown of the EWOD electrode can be prevented. That is, according to the present embodiment, the transport apparatus 500 including the EWOD electrode 300 can be used a plurality of times for extracting nucleic acid into the capillary array 114.

Example 3

In example 1 and example 2, the case where the transport device 500 held horizontally is tilted by a load and the sample droplet 502 is moved to the sample extracting unit 306 is described. In order to move the sample droplet 502 to the sample extraction unit 306, the sample extraction unit 306 may be located at a position lower than the droplet channel of the EWOD electrode 300A having the end, and the two may be smoothly connected to each other. Therefore, in the present embodiment, a configuration in which the conveying device 500 is tilted instead of tilting the conveying device 500 kept horizontal by a load will be described.

This embodiment will be described with reference to fig. 12. Fig. 12 is an XZ sectional view of the conveyance device 500, as in fig. 5 (a). The transport facility 500 of the present embodiment includes an upper plate 401, an upper electrode 402, an upper water repellent film 403, a lower water repellent film 405, an insulating film 406, an EWOD electrode 300, a lower plate 407, an opening 501, and a droplet retention section 1001 as in embodiment 2. Further, a trapezoidal support portion 1200 is provided between the transport facility 500 and the biochemical cartridge 121. The trapezoidal support portion 1200 may be provided between the biochemical cartridge 121 and the movable stage 123. The upper plate 401, the upper electrode 402, the upper water repellent film 403, the lower water repellent film 405, the insulating film 406, the EWOD electrode 300, the lower plate 407, the opening 501, and the droplet holding portion 1001 are the same as those in example 2, and therefore, the description thereof is omitted.

The trapezoidal support portion 1200 is provided at an end portion of the transport facility 500 in the-X direction, is a member that supports the transport facility 500 so as to be inclined, and has a trapezoidal shape in the XZ cross section. By inclining the transport device 500, the sample extraction unit 306 is located at a position lower than the droplet channel of the EWOD electrode 300A having the end, and the space between the droplet channel and the sample extraction unit 306 becomes an inclined surface.

The operation of the present embodiment will be described with reference to fig. 13.

(S1301)

By controlling the voltage applied to the EWOD electrode 300 of the transport apparatus 500, the sample droplet 502 is transported to the position of the end EWOD electrode 300A. The transport device 500 is tilted to generate a driving force in the direction of the arrow in fig. 12 on the sample droplet 502, but the EWOD electrode 300 is held at the position of the distal EWOD electrode 300A because a voltage is applied to the EWOD electrode 300.

(S1302)

The applied voltage to the EWOD electrode 300 is set to an off or ground state. When the applied voltage is turned off or grounded, the force holding the sample droplet 502 at the position of the EWOD electrode 300A at the end disappears, and therefore the sample droplet 502 starts moving in the direction of the arrow in fig. 12. The sample droplet 502 moves along the upper surface of the lower water repellent film 405, and is retained at the position of the sample extraction unit 306 by the droplet retaining unit 1001.

(S1303)

The capillary array 114 moves in the + Z direction, and is inserted into the transport device 500 through the opening 501, and the cathode end 126 of the capillary 102 comes into contact with the sample droplet 502.

(S1304)

The nucleic acid in the sample droplet 502 is extracted to the capillary array 114 by applying a voltage of several kV to the loading head 124. After the nucleic acid in the sample droplet 502 is extracted by a predetermined amount into the capillary array 114, the voltage application to the loading head 124 is turned off.

(S1305)

The capillary array 114 moves in the-Z direction and the droplet holding portion 1001 slides in the Z direction or the Y direction, whereby the sample droplet 502 remaining after the nucleic acid is extracted into the capillary array 114 moves on the inclined surface and is discharged from the transport device 500.

According to the present embodiment described above, in the sample extracting section 306 provided at a position having a predetermined distance from the EWOD electrode 300A at the end of the transport apparatus 500, the nucleic acid in the sample droplet 502 can be extracted to the capillary array 114. Since the sample extraction unit 306 is located at a position where electrical insulation can be maintained even when a voltage necessary for extracting nucleic acid by the capillary array 114 is applied to the loading head 124, dielectric breakdown of the EWOD electrode can be prevented. That is, according to the present embodiment, the transport apparatus 500 including the EWOD electrode 300 can be used a plurality of times when extracting nucleic acid using the capillary array 114.

In addition, according to the present embodiment, since the driving device such as the elevating unit 1000 is not provided, the configuration can be simplified as compared with embodiment 2.

Example 4

In embodiment 3, a structure in which the entire conveying apparatus 500 is inclined is described. In order to move the sample droplet 502 to the sample extraction unit 306, the sample extraction unit 306 may be located at a position lower than the droplet channel of the EWOD electrode 300A having the end, and may have a smoothly continuous surface therebetween. Therefore, in the present embodiment, the following configuration is explained: instead of inclining the entire transport device 500, a part of the transport device 500 is kept horizontal, and an inclined surface inclined with respect to the horizontal plane is provided between the droplet flow path and the sample extracting unit 306.

This embodiment will be described with reference to fig. 14. Fig. 14 is an XZ sectional view of the conveyance device 500, as in fig. 5 (a). The transport facility 500 of the present embodiment includes an upper plate 401, an upper electrode 402, an upper water repellent film 403, a lower water repellent film 405, an insulating film 406, an EWOD electrode 300, a lower plate 407, an opening 501, and a droplet retention section 1001 as in embodiment 2. In addition, a first supporting portion 1400 and a second supporting portion 1401 are provided between the transport apparatus 500 and the biochemical cartridge 121. The upper electrode 402, EWOD electrode 300, opening 501, and droplet holding portion 1001 are the same as those in example 2, and therefore, the description thereof is omitted.

The upper plate 401, the upper water repellent film 403, the lower water repellent film 405, the insulating film 406, and the lower plate 407 have a structure in which they are smoothly bent at the position of the EWOD electrode 300A at the end. That is, the upper plate 401, the upper water repellent film 403, the lower water repellent film 405, the insulating film 406, and the lower plate 407 are kept horizontal in a region where the upper electrode 402 and the EWOD electrode 300 are provided, and have inclined surfaces outside the region. By providing the transport device 500 with such an inclined surface, the sample collection unit 306 is disposed at a position lower than the droplet flow path of the EWOD electrode 300A having the end. The droplet channel and the sample collection unit 306 are smoothly connected to each other through the lower water repellent film 405. The space between the droplet flow path and the sample collection unit 306 is not limited to the plane shown in fig. 14, and may be a curved surface smoothly connected to each other.

The first support 1400 and the second support 1401 are members that support the conveying apparatus 500. The first support 1400 is provided at an end of the transport apparatus 500 in the-X direction, and the second support 1401 is provided at a position where the transport apparatus 500 is smoothly bent.

The operation of this embodiment is the same as that of embodiment 3, and follows the flowchart shown in fig. 13, and therefore, the description thereof is omitted.

According to the present embodiment described above, in the sample extracting section 306 provided at a position having a predetermined distance from the EWOD electrode 300A at the end of the transport apparatus 500, the nucleic acid in the sample droplet 502 can be extracted to the capillary array 114. Since the sample extraction unit 306 is located at a position where electrical insulation can be maintained even when a voltage necessary for extracting nucleic acid by the capillary array 114 is applied to the loading head 124, dielectric breakdown of the EWOD electrode can be prevented. That is, according to the present embodiment, the transport apparatus 500 including the EWOD electrode 300 can be used a plurality of times when extracting nucleic acid using the capillary array 114.

In addition, according to the present embodiment, since the portion having the EWOD electrode 300 is kept horizontal and the process until the sample droplet 502 is obtained can be free from the influence of gravity, the control of the EWOD electrode 300 is easier than that of embodiment 3.

The biochemical analyzer according to the present invention is not limited to the above-described embodiments, and constituent elements may be modified and embodied within a range not departing from the gist of the present invention. Further, a plurality of constituent elements disclosed in the above embodiments may be appropriately combined. Further, several components may be deleted from all the components shown in the above embodiments.

Description of the symbols

101: device main body, 102: capillary, 103: pump mechanism, 104: high-voltage power supply, 105: syringe, 106: gel block, 107: separation medium container, 108: anode buffer container, 109: anode electrode, 110: electric valve, 111: light source, 112: optical detector, 113: detection unit, 114: capillary array, 115: thermostatic bath, 116: fan, 117: heater, 118: buffer container, 119: cleaning container, 120: waste liquid container, 121: cartridge for biochemistry, 122: conveyor, 123: mobile station, 124: loading head, 125: control computer, 126: cathode terminal, 127: control substrate, 128: optical data, 129: capillary head, 300: EWOD electrode, 301: sample cell, 302: reagent tank, 303: sample flow path, 304: reagent flow path, 305: temperature control zone, 306: sample extraction unit, 400: droplet, 401: upper plate, 402: upper electrode, 403: upper water repellent film, 404: fluid, 405: lower water repellent film, 406: insulating film, 407: lower plate, 500: conveying equipment, 501: opening, 502: sample droplet, 503: fulcrum portion, 504: support portion, 505: rut, 1000: lifting unit, 1001: droplet retention portion, 1200: trapezoidal support, 1400: first support portion, 1401: a second support portion.