A biochip, a detection method and a device for automatic solid sample adding

Abstract

The application discloses a kind of solid sample automatic sample adding biochip, detection method and detection device, biochip includes biochip substrate and detection area, further include container and cavity, the container is fragile material, the cavity is the hollow cavity with top opening, the lower portion of the cavity is communicated with sample introduction microtube line, another end of the sample introduction microtube line is communicated with the detection area, the side wall of the cavity is provided with breakage opening, the bottle mouth of the container is provided with plunger with interference fit, the maximum outer diameter of the plunger is greater than the maximum outer diameter of the container, the plunger is interference fit with the inner wall of the cavity, the container and the breakage opening are located at the same horizontal position.By the design of the application, after the container is broken, the container fragments and freeze-dried powder will all fall to the bottom of the cavity under the weight, automatic addition is realized.Freeze-dried reagent storage requires high, after being stored separately with container, the air-tightness requirement of transfer cavity and substrate is reduced.

Description

Technical Field

[0001] This invention relates to biochips, and in particular to a biochip, detection method, and detection device for automatic solid sample dispensing. Background Technology

[0002] Currently, nucleic acid testing methods use reagents in liquid form, stored at -20°C. During testing, the reagents are manually or automatically added to the instrument or biochip. Using liquid reagents requires storage at -20°C, placing stringent requirements on storage and transportation, and making it difficult to individually aliquot into biochips. Furthermore, using liquid reagents necessitates manual addition by healthcare professionals when using the biochip, hindering automated operation.

[0003] If liquid reagents are freeze-dried, they only need to be stored in a conventional refrigerated environment at 2-8°C. While ensuring the freeze-dried reagents are preserved with an inert gas, it is necessary to solve the problem of easily assembling these reagents into a biochip, using simple equipment, requiring no manual intervention from medical personnel, and enabling automated sample addition of freeze-dried powder. Based on this technical background, this application designs a biochip for automated solid sample addition. The freeze-dried reagents are pre-freeze-dried and stored in glass vials, which are stored separately. After the biochip is assembled, the glass vial containing the freeze-dried reagent is directly assembled into the biochip. When the biochip is used, the glass vial is broken, and the freeze-dried powder falls into the designed cavity, achieving automated sample addition.

[0004] Patent CN106461535A discloses a reagent kit and method for cell detection, including a housing and an actuator. The housing includes a puncture member defining a transfer path in fluid communication with a reagent volume. When the housing is coupled to a reaction chamber, a delivery portion of the housing defines a delivery path between the transfer path and the reaction chamber. The actuator has a plunger portion disposed within the reagent volume. An engaging portion of the actuator can be manipulated to move the plunger portion within the reagent volume, thereby deforming the reagent container. The puncture member can puncture a fragile portion of the reagent container to transfer reagent from the container via the transfer path and / or delivery path into the reaction chamber. This patent delivers reagents through tubing and is suitable for adding liquid reagents. Due to limitations of the delivery tubing and the fact that the puncture member can puncture a fragile portion of the reagent container, powder residue would remain in the reagent container if used for adding solid powder; therefore, this patent is not applicable to the quantitative addition of solid powder. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide a biochip, detection method and detection device for automatic addition of solid samples, which addresses the shortcomings of the existing technology and realizes automatic quantitative addition of solid powder reagents that need to be sealed and preserved.

[0006] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is: a biochip for automatic solid sample dispensing, comprising a biochip substrate and a detection area, and further comprising a container and a cavity. The container includes a fragile part, and the cavity is a hollow cavity with a top opening. The lower part of the cavity is connected to a sample inlet microchannel, the other end of which is connected to the detection area. A breakage port is provided through the side wall of the cavity. A plunger is provided with an interference fit at the mouth of the container. The maximum outer diameter of the plunger is larger than the maximum outer diameter of the container. The container is assembled into the cavity through the top opening of the cavity. The plunger is interference fitted with the inner wall of the cavity. The fragile part and the breakage port are located at the same horizontal position.

[0007] The freeze-dried powdered reagent is placed in a container and sealed with a plunger for secure storage. The container is then placed inside the chamber; when testing is required, the thin-walled portion of the container is broken, allowing for automatic sample dispensing. Fragile materials include glass, ceramics, and porcelain. The plunger is made of butyl-based rubber, known for its excellent sealing properties.

[0008] When the container and plunger are loaded into the sample dispensing device, the maximum outer diameter of the plunger is greater than the maximum outer diameter of the container, which can achieve an interference fit between the plunger and the sample dispensing device, making it easier for the container to move in the sample dispensing device.

[0009] Furthermore, the cavity includes a cylindrical cavity and a constricted opening at the bottom of the cylindrical cavity. The cylindrical cavity and the constricted opening are connected by an inclined surface. The sample injection microchannel is horizontally positioned and communicates with the lower sidewall of the constricted opening. Large container fragments remain above the constricted opening, while smaller fragments are generated in smaller quantities and enter the bottom of the constricted opening, preventing them from entering the detection area through the sample injection microchannel.

[0010] Furthermore, the container is a glass bottle, comprising a thick-walled portion near the container opening and a thin-walled portion near the bottom of the container. The wall thickness of the thick-walled portion is 1-2 mm, and the wall thickness of the thin-walled portion is 0.3-1 mm. The thin-walled portion is at the same horizontal level as the breakage opening. To directly load the reagent into the container for freeze-drying, glass is chosen as the container material, as glass bottles are well-suited for freeze-drying. To achieve a balance between glass bottle breakage and freeze-drying efficiency, glass bottles of different thicknesses, ranging from 0.3 to 1 mm, are selected.

[0011] Furthermore, when the wall thickness of the thin-walled portion is 0.5~1mm, a horizontal annular pre-cut line is formed along the outer wall of the thin-walled portion. For glass bottles with a thickness of 0.5mm or more, the glass bottle will be pre-cut as needed to make the glass bottle easier to break and to facilitate maintaining the consistency of the broken glass fragments.

[0012] Furthermore, the distance between the sample injection microchannel and the bottom of the constriction is 1-2 mm. This results in minimal glass fragments, which settle at the bottom of the constriction and do not enter the detection area with the sample injection microchannel.

[0013] Furthermore, the plunger includes an insertion portion and an upper plunger portion, the diameter of which is larger than the maximum outer diameter of the container. The insertion portion achieves an interference fit with the container opening, and the upper plunger portion achieves an interference fit with the inner wall of the cavity.

[0014] The insert portion has multiple notches on its circumference. These notches facilitate air permeability during freeze-drying, allowing the solution to be directly contained in the container for freeze-drying. During freeze-drying, the plunger is not sealed inside the container, facilitating the freeze-drying of the solution. After freeze-drying, the plunger is pressed down to ensure an interference fit between the insert portion and the inner wall of the container.

[0015] Furthermore, the biochip also includes a reactor with a liquid outlet at the top, and an inlet is provided on the side wall of the cavity. The liquid outlet of the reactor is located adjacent to the inlet. After adding the freeze-dried reagent in the container, other reagents are added through the inlet. By tilting the biochip, the liquid in the reactor can be poured into the cavity through the inlet to mix and dissolve with the freeze-dried powder in the cavity. After adding the reagents, the plunger is pressed down to seal the inlet, keeping the lower part of the cavity containing the reagents sealed.

[0016] Furthermore, the height of the plunger is greater than the diameter of the break-in port, and the height of the plunger is greater than the diameter of the injection port. After the container is broken, the plunger is pressed down to seal the break-in port. After the liquid sample is added, the plunger is pressed down to seal the break-in port.

[0017] This invention also discloses a detection method for a biochip, comprising the following steps:

[0018] S1. Insert a plunger into the container containing the freeze-drying reagent, and put the container and plunger into the cavity through the upper part of the cavity, with the container and the breakout port at the same level.

[0019] S2. Use a flat-headed rod to strike the container, causing it to break. The broken container fragments and freeze-drying reagent fall to the bottom of the chamber under their own weight. The diameter of the flat-headed rod is 1-3 mm smaller than the diameter of the container.

[0020] S3. Press down the plunger to seal the breakout port;

[0021] S4. After completing the liquid injection, dissolve and freeze-dry the reagent to obtain a mixed liquid. Extract the mixed liquid through the injection microtube for detection.

[0022] Using a flat-headed rod to impact the container, with the rod's diameter being 1-3 mm smaller than the container's diameter, can ensure that the container breaks evenly and produces less debris.

[0023] The present invention also discloses a detection device for a biochip, wherein a flat-headed rod is horizontally mounted on the detection device, and the flat-headed rod and the breakout port are located at the same horizontal position.

[0024] To facilitate later use of lyophilized reagents, the smaller the fragment size of the broken container, the better. Small fragment size can be obtained through the following settings:

[0025] 1. Container installation height: The bottom of the container should be flush with the bottom of the rupture hole.

[0026] 2. Depth of the flat-head rod into the chip: The flat-head rod continues to advance 1-3mm after contacting the freeze-drying container until it breaks.

[0027] 3. The diameter of the pressure rod is set according to the glass freeze-drying container.

[0028] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0029] 1. Through the design of this invention, after the container is broken, the container fragments and freeze-dried powder will fall to the bottom of the cavity under their own weight, thus achieving automatic replenishment;

[0030] 2. Because freeze-dried reagents have high preservation requirements, storing them separately in containers simplifies the biochip design and reduces the airtightness requirements between the transfer chamber and the biochip substrate.

[0031] 3. In this invention, the cap plunger portion of the container is thicker to prevent the plunger from cracking, while the lower portion is thinner, which is beneficial for crushing and adding samples.

[0032] 4. Freeze-dried powder adhering to the side wall of the cavity can also be pushed to a specific position by the plunger.

[0033] 5. After adding the freeze-dried powder, add liquid reagent to dissolve the freeze-dried powder, thus achieving quantitative addition of the freeze-dried powder. Attached Figure Description

[0034] Figure 1 This is a front view of a biochip in one embodiment of the present invention;

[0035] Figure 2 for Figure 1 AA section view;

[0036] Figure 3 for Figure 1 Top view;

[0037] Figure 4This is a front view of an automatic sampling device according to an embodiment of the present invention;

[0038] Figure 5 for Figure 4 CC section view;

[0039] Figure 6 for Figure 4 Top view;

[0040] Figure 7 This is a front view of a container in one embodiment of the present invention;

[0041] Figure 8 for Figure 7 BB section view;

[0042] Figure 9 This is a schematic diagram of the three-dimensional structure of the plunger.

[0043] The components are as follows: 1. Biochip substrate; 2. Breakout port; 3. Cavity; 31. Columnar cavity; 32. Narrowing; 33. Sloping surface; 4. Detection area; 5. Reactor; 6. Sample inlet; 7. Mixing area; 8. Assembly port; 9. Plunger; 91. Insertion part; 92. Upper part of plunger; 93. Notch; 10. Container; 101. Thick-walled part; 102. Thin-walled part; 11. Breakout device; 12. Freeze-dried reagent; 13. Normally closed laser valve; 14. Sample inlet microchannel; 15. Top opening; 16. Pre-filled reagent area one; 17. Pre-filled reagent area two; 18. Waste liquid storage area. Detailed Implementation

[0044] like Figures 1-3 As shown, the biochip includes a biochip substrate 1, a detection zone 4, a reactor 5, an automatic sample dispenser, a pre-filled reagent zone 16, a pre-filled reagent zone 17, and a waste liquid storage zone 18. The biochip substrate 1 has an assembly port 8. The automatic sample dispenser is installed below the biochip substrate 1, and the assembly port 8 matches the top opening 15 of the cavity 3. The cavity 3 is assembled onto the biochip substrate 1 via a normally closed laser valve 13, which is integrally injection molded with the biochip substrate 1. The normally closed laser valve 13 is connected to one end of an injection microchannel 14, and the other end of the injection microchannel 14 is connected to the detection zone 4. The reactor 5 has an outlet at the top, and the cavity 3 has an inlet 6 on its side wall, with the outlet of the reactor 5 located adjacent to the inlet 6. The detection zone 4 is equipped with a vacuum drive device, which is connected to the injection microchannel 14.

[0045] The best way to open a normally closed microvalve is to use laser to heat the valve core of the normally closed laser valve at a specific point. This melts the valve core of the normally closed laser valve, thereby creating a passage between the sample injection microchannel and the cavity.

[0046] like Figures 4-6As shown, the automatic sample dispensing device includes a container 10 and a cavity 3. The cavity 3 is a hollow cavity with an open top. The bottom of the cavity 3 is connected to a sample inlet microtube 14. A break-out port 2 is provided through the side wall of the cavity 3. The container 10 is assembled into the cavity 3 through the top opening.

[0047] like Figures 7-8 As shown, in this embodiment, container 10 is a glass bottle, which is a cylindrical bottle with an open top. Container 10 is used to hold freeze-dried reagent 12 and mainly consists of container 10 and plunger 9. The glass bottle has a thin-walled lower section and a thick-walled plunger section. The thin-walled section has a wall thickness of 0.5 mm, and the thick-walled section has a wall thickness of 1 mm. The outer diameter of the glass bottle is 5 mm, and the volume is 100 μL. When the wall thickness of the thin-walled section 102 is 0.5~1 mm, a horizontal annular pre-cut line is formed along the outer wall of the thin-walled section 102. The diameter of the container is between 5~10 mm, and the volume is between 100~1000 μL. The maximum outer diameter of the plunger is slightly larger than the maximum outer diameter of the container. The plunger 9 includes an insertion part 91 and an upper plunger part 92. The diameter of the upper plunger part 92 is larger than the maximum outer diameter of the container 10. The insertion part 91 has multiple notches formed around its circumference. The container 10 and the plunger 9 are fitted with an interference fit. The plunger material is a butyl rubber material with good sealing properties. The reagent is freeze-dried and stored separately.

[0048] During freeze-drying, first, load the freeze-drying reagent 12 into container 10, then place the glass vial containing the reagent into a vacuum freeze dryer for freeze-drying. Next, introduce a protective gas, such as nitrogen, into the glass vial. Finally, cap it with plunger 9 and store it separately.

[0049] The maximum outer diameter of plunger 9 is greater than the maximum outer diameter of container 10. Plunger 9 is interference-fitted with cavity 3. Plunger 9 can move vertically within cavity 3 under external force. Thin-walled portion 102 is at the same horizontal level as breakout port 2. The length of plunger 9 is greater than the diameter of breakout port 2. A sample inlet 6 is provided through the side wall of cavity 3, located below breakout port 2. The height of plunger 9 is greater than the diameter of sample inlet 6. The diameter of sample inlet microchannel 14 is 0.5 mm. The lower part of the cavity is a mixing zone 7, which is equipped with a mixing device. The mixing device can be an ultrasonic mixing mechanism, a mechanical vibration mechanism, or a rotation mechanism.

[0050] The cavity 3 includes a cylindrical cavity 31 and a constricted opening 32 at the bottom of the cylindrical cavity 31. The cylindrical cavity 31 and the constricted opening 32 are connected by an inclined surface 33. The sample injection microchannel 14 is horizontally arranged and communicates with the lower side wall of the constricted opening 32. The constricted opening 32 is a flat cuboid. The distance between the sample injection microchannel 14 and the bottom of the constricted opening 32 is 1-2 mm.

[0051] When the biochip is assembled into a finished product, the bottle containing the freeze-dried reagent is then automatically assembled through assembly port 8. The cavity 3 is made of plastic and is integrally injection molded, relying on the interference fit between the plunger 9 and the cavity 3 for assembly and positioning. A certain gap exists between the container 10 and the cavity 3. The maximum outer diameter of the plunger is slightly larger than the maximum outer diameter of the glass bottle, and it is in an interference fit with the cavity 3. The crushing device 11 uses a motor with a flat-head pressure rod, and the force of the pressure rod is 8-20 kg. The diameter of the flat-head pressure rod of the crushing device 11 is slightly smaller than the diameter of the glass bottle, by 2 mm. Crushing with a flat-head rod of similar diameter results in relatively uniform glass fragments after crushing, without any large individual pieces.

[0052] When using the biochip, the container 10 is first broken by the breaking device 11. The broken glass fragments and freeze-dried reagent 12 fall to the bottom of the chamber 3 under their own weight. Then, the plunger 9 is pressed down to seal the breaking port 2. Next, the pre-reacted reagent in the reactor 5 is poured into the chamber 3 through the sample inlet 6 by rotating the plunger. After pouring, the biochip is rotated back to its original position. During this process, the reagent falls back to the bottom of the chamber 3. The plunger 9 is then pressed down again until the sample inlet 6 is sealed. The mixed liquid is mixed in the mixing zone 7 by vibration / ultrasound. After the mixed liquid is fully mixed, the normally closed laser valve 13 is opened, and the mixed liquid enters the detection zone 4 through the sample injection microchannel 14 under vacuum to realize the detection of the target substance of the biochip.

Claims

1. A biochip for automatic solid sample dispensing, comprising a biochip substrate (1) and a detection area (4), characterized in that, It also includes a container (10) and a cavity (3). The container (10) includes a fragile part. The cavity (3) is a hollow cavity with an open top. The lower part of the cavity (3) is connected to a sample inlet microtube (14). The other end of the sample inlet microtube (14) is connected to the detection area (4). A breakage port (2) is provided through the side wall of the cavity (3). The bottle mouth of the container (10) is fitted with a plunger (9). The maximum outer diameter of the plunger (9) is greater than the maximum outer diameter of the container (10). The container (10) is assembled into the cavity (3) through the top opening of the cavity (3). The plunger (9) is fitted with the inner wall of the cavity (3). The fragile part and the breakage port (2) are located at the same horizontal position. The container (10) is a glass bottle. The container (10) includes a thick-walled part (101) near the opening of the container and a thin-walled part (102) near the bottom of the container. The wall thickness of the thick-walled part (101) is 1~2mm, and the wall thickness of the thin-walled part (102) is 0.3~1mm. The thin-walled part (102) is at the same horizontal position as the break opening (2).

2. The biochip according to claim 1, characterized in that, The cavity (3) includes a columnar cavity (31) and a constriction (32) at the bottom of the columnar cavity (31). The columnar cavity (31) and the bottom constriction (32) are connected by an inclined surface (33). The sample injection microchannel (14) is horizontally arranged and communicates with the lower side wall of the constriction (32).

3. The biochip according to claim 1, characterized in that, When the wall thickness of the thin-walled portion (102) is 0.5~1mm, a horizontal annular pre-cut line is provided along the outer wall of the thin-walled portion (102).

4. The biochip according to claim 2, characterized in that, The distance between the sample injection microchannel (14) and the bottom of the constriction (32) is 1-2 mm.

5. The biochip according to any one of claims 1-4, characterized in that, The plunger (9) includes an insertion portion (91) and an upper plunger portion (92), the diameter of which is greater than the maximum outer diameter of the container (10).

6. The biochip according to any one of claims 1-4, characterized in that, The biochip also includes a reactor (5) with an outlet at the top, and an inlet (6) is provided on the side wall of the cavity (3). The outlet of the reactor (5) is located near the inlet (6).

7. The biochip according to claim 5, characterized in that, The height of the plunger (9) is greater than the diameter of the breakout port (2), and the height of the plunger (9) is greater than the diameter of the injection port (6).

8. A detection method for a biochip according to any one of claims 1-7, characterized in that, Includes the following steps: S1. Insert a plunger into the container containing the freeze-drying reagent, and put the container and plunger into the cavity through the upper part of the cavity, with the container and the breakout port at the same level. S2. Use a flat-headed rod to strike the container, causing it to break. The broken container fragments and freeze-drying reagent fall to the bottom of the chamber under their own weight. The diameter of the flat-headed rod is 1-3 mm smaller than the diameter of the container. S3. Press down the plunger to seal the breakout port; S4. After completing the liquid injection, dissolve and freeze-dry the reagent to obtain a mixed liquid. Extract the mixed liquid through the injection microtube for detection.

9. A detection device for a biochip according to any one of claims 1-7, characterized in that, A flat-headed rod is horizontally installed on the detection device, and the flat-headed rod and the crushing port (2) are located at the same horizontal position.

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