CN117160548A

CN117160548A - Microfluidic chip and related device and method

Info

Publication number: CN117160548A
Application number: CN202210593913.3A
Authority: CN
Inventors: 王健; 李宁波; 杜文超; 孟庆亮; 陈婵平
Original assignee: Qingdao Huada Zhizao Jichuang Technology Co ltd
Current assignee: Qingdao Huada Zhizao Jichuang Technology Co ltd
Priority date: 2022-05-27
Filing date: 2022-05-27
Publication date: 2023-12-05

Abstract

The invention relates to the technical field of display, and provides a microfluidic chip and a detection device comprising the microfluidic chip, wherein the microfluidic chip at least comprises a substrate and a sensing device which are oppositely arranged, a driving electrode is arranged on the substrate, and a channel for accommodating liquid drops to be detected is formed between the substrate and the sensing device; the sensing device comprises a control part and a sensing part which are connected with each other, wherein the control part can control at least one of the sensing part and the substrate to move relatively towards a direction approaching to or away from each other so as to enable the sensing part to form an electric connection path with the driving electrode through the liquid drop to be detected. The detection device provided by the invention can accurately control the distance between the sensing component and the liquid drops to be detected with different sizes, thereby improving the precision of the test data; and when the microfluidic chip adopts a sealing structure, the volatilization rate of liquid drops can be effectively slowed down.

Description

Microfluidic chip and related device and method

Technical Field

The invention relates to the technical field of display, in particular to a microfluidic chip, a detection device comprising the microfluidic chip and a detection method.

Background

Gene sequencing is a hotspot in the current biomedical field, and in sequencing engineering, it is particularly important to study single-cell or biochemical droplet systems. The existing water drop angle detection device can detect water drop angles or dynamic water drop angles on different base materials aiming at water drops or liquid drops, so that the hydrophilic and hydrophobic properties are quantitatively evaluated, and the device is widely applied to testing and research in the fields of silicon crystals, liquid crystals, glass, fibers, synthetic materials and the like. Compared with the traditional microarray and microfluidic system, the digital microfluidic device has stronger operability, does not need an external gas circuit device, can realize automation and micromation, and is widely applied to the water drop angle detection device.

The existing digital microfluidic devices are mostly open monopolar plates and bipolar plates under single conditions. The monopole type is driven by a surface-treated metal wire as an electrode, water drops or liquid drops on a chip electrode move, and video is recorded by a camera, so that the movement performance of the water drops or the liquid drops is evaluated; the bipolar plate type is monitored by a camera to evaluate the movement performance of the water drops or the liquid drops and obtain the water drop angle size of the liquid drops. However, the upper electrode is fixedly arranged and the position of the upper electrode cannot be adjusted no matter the upper electrode is of a monopolar plate type or a bipolar plate type digital microfluidic device, so that the distance between the upper electrode and liquid drops with different sizes is difficult to ensure, and the accuracy of test data is not improved.

In view of this, the present invention has been made.

Disclosure of Invention

The invention provides a continuous dynamic detection device for digital liquid drops for a sequencer, which aims to solve the problems that in the prior art, the distance between an upper electrode and liquid drops with different sizes is difficult to ensure and the improvement of the precision of test data is not facilitated due to the fixed position of the upper electrode.

In order to solve the above problems, the present invention provides in a first aspect a microfluidic chip, which at least includes a substrate and a sensing device disposed opposite to each other, wherein a driving electrode is disposed on the substrate; a channel for accommodating the liquid drop to be detected is formed between the substrate and the sensing device; the sensing device comprises a control part and a sensing part which are connected with each other, wherein the control part can control at least one of the sensing part and the substrate to move relatively towards a direction approaching to or away from each other so as to enable the sensing part to form an electric connection path with the driving electrode through the liquid drop to be detected.

The microfluidic chip at least comprises a substrate and a sensing device, wherein a driving electrode is arranged on the substrate, a channel for accommodating liquid drops to be detected is formed between the substrate and the sensing device, and the liquid drops to be detected with different shapes and components can be dripped into the channel above the driving electrode; the sensing device comprises a control component and a sensing component which are connected with each other, the control component can control at least one of the sensing component and the substrate to move relatively towards the direction which is close to or far away from each other, namely, the sensing component can be controlled to move or the substrate can be controlled to move, so that the sensing component can form an electric connection path with the driving electrode through the liquid drop to be tested, the sensing component can sense the shape and the size of the liquid drop to be tested, the sensed information (such as the interval between the liquid drop to be tested and the sensing component) is fed back to the control component, the control component can adjust the position of the sensing component (or the substrate) up and down according to the information, so that the interval between the sensing component (namely, the upper electrode) and the liquid drop to be tested with different sizes can be controlled accurately, and the accuracy of test data is improved.

In other preferred embodiments, the microfluidic chip further includes a sidewall connected to the top plate and extending toward the substrate; the sensing component comprises a top plate and at least one sensing electrode arranged on the top plate; the side wall, the top plate and the base plate form a sealed cavity, and when the control part controls the top plate to drive the sensing electrode to move, the top plate changes in position relative to the base plate in the height direction of the side wall; the drive electrode is at least partially located within the sealed cavity.

When the liquid drop to be measured is in a sealed environment, the volatilization problem of the liquid drop can be effectively solved, particularly in the process of voltage transformation and detection of rapid liquid drop volatilization, the volatilization reducing effect is more obvious, at least one sensing electrode is arranged on the top plate, the number of the sensing electrodes which can work is matched with the number and the positions of the liquid drops to be measured in a one-to-one correspondence mode, when the control part controls the top plate to drive the sensing electrodes to move, the top plate changes in position relative to the substrate in the height direction of the side wall, the vertical distance between the sensing electrodes and the liquid drops to be measured can be adjusted, the distance between the upper electrode (namely the sensing electrodes) and the liquid drops to be measured with different sizes can be accurately controlled, the driving electrode is at least partially positioned in the sealed cavity, the liquid drops to be measured can be placed above the part of the driving electrode positioned in the sealed cavity, and the sensing electrodes can form an electric connection path with the driving electrode through the liquid drops to be measured.

In other preferred embodiments, at least one driving electrode is disposed on the upper surface of the substrate, at least one detection area is disposed on the upper surface of the driving electrode, at least one droplet to be detected can be dropped in the detection area, and the sensing electrode corresponds to the droplet to be detected in the detection area below the sensing electrode one by one; the sensing electrodes are arranged in one-to-one correspondence with the detection areas, the sensing electrodes are generally positioned above the detection areas corresponding to the sensing electrodes, and the microfluidic chip further comprises a positioning device for positioning one side, close to the detection areas, of the sensing electrodes, the distance between the sensing electrodes and the liquid drops can be monitored through the positioning device, and the distance between the sensing electrodes and the liquid drops to be detected with different sizes can be conveniently adjusted.

In other preferred embodiments, all sensing electrodes are movably disposed on the top plate and are all connected to the control member; the control part further controls the sensing electrode to move towards or away from the liquid drop to be detected relative to the top plate on the basis of the movement of the top plate relative to the base plate.

When the detection is carried out, after the liquid drop to be detected is dripped in a certain detection area, the control part can control the sensing electrode at the position corresponding to the liquid drop to be detected to stretch out, then the control part can move on the basis of the movement of the top plate relative to the substrate (namely, only the top plate can move), after the top plate reaches the initial determined position (coarse adjustment), the sensing electrode is further controlled to move towards the direction close to or far away from the liquid drop to be detected (fine adjustment) relative to the top plate, and the tip of the sensing electrode can be contacted with the liquid drop to be detected through the cooperation of the coarse adjustment and the fine adjustment, so that the detection of the contact angle of the liquid drop is facilitated.

In other preferred embodiments, the side wall is provided with a light-transmitting portion through which light passes, and the light transmittance of the light-transmitting portion is greater than or equal to 0.8, so as to ensure a better light-transmitting effect, the external light source makes light incident into the sealed cavity through the light-transmitting portion, and the light-transmitting portion is highly transparent, so that light penetration and image acquisition are facilitated.

In other preferred embodiments, the side wall includes an inner wall of transparent material and an outer wall of opaque material, and the light-transmitting portion is disposed on the outer wall.

The inner wall and the outer wall in this scheme can directly laminate and set up, also can leave the space, are provided with printing opacity portion on opaque outer wall, and external light source is through printing opacity portion and transparent inner wall with light incidence to sealed intracavity, the follow-up image acquisition work's of being convenient for normal clear.

In other preferred embodiments, the sensing electrode comprises a wire electrode and a shaft sleeve axially sleeved on the periphery of the wire electrode, the shaft sleeve drives the wire electrode to be movably connected with the top plate, and the shaft sleeve can effectively protect the wire electrode embedded therein, so that the sensing electrode can normally and stably exert a sensing effect.

In other preferred embodiments, the driving electrode includes a patterned electrode sheet, and a first protective layer disposed on a side of the electrode sheet near the top plate; the first protective layer comprises at least a first dielectric layer and a first hydrophobic layer, and the driving electrode is used for controlling the movement of the liquid drops. The upper surface of the electrode plate needs to be provided with a dielectric layer and a hydrophobic layer, or the two-in-one dielectric hydrophobic layer can be realized by means of film pasting, spraying, deposition and the like, and the dielectric layer is used for maintaining the insulativity between the circuit and each layer.

In another preferred scheme, a second protective layer is arranged on one side of the top plate, which is close to the substrate; the second protective layer at least comprises a conductive layer and a second hydrophobic layer positioned on one side of the conductive layer near the substrate. The lower surface of the top plate may be structured with a conductive layer, such as by spraying, depositing, sputtering, or the like, or with a conductive material, and the conductive layer may be structured with a hydrophobic layer, or with a conductive and hydrophobic material.

In other preferred embodiments, the device further comprises a temperature control component connected to the substrate, wherein the temperature control component is used for adjusting the temperature of the substrate to control the temperature in the sealing cavity to be 4-97 ℃, so that the device not only can solve the volatilization problem of the existing drop angle testing device, but also can evaluate and detect the heating state and the PCR process of drops.

The second aspect of the invention provides a detection device comprising the microfluidic chip, the detection device further comprising a light source; and

an output module including a first output terminal connected to the driving electrode and a second output terminal connected to the sensing part; the output module can output different voltage driving signals to the electric connection path;

The image processing module is used for collecting the morphology of the liquid drops in the channels under different voltage driving signals and detecting the contact angle of the liquid drops.

When the detection device is specifically applied to detect the contact angle of the liquid drop to be detected, the light source is aligned to the light transmitting part, so that light enters the cavity and is aligned to the liquid drop to be detected, the contact angle is conveniently detected, the position of the sensing electrode is regulated to be in contact with the liquid drop to be detected, the output module is opened, the driving electrode and the sensing electrode are subjected to pressure supply, different voltage driving signals are provided for the electric connecting passage, finally, the morphology of the liquid drop in the passage under the different voltage driving signals is collected through the image processing module, and the contact angle of the liquid drop to be detected is detected.

In other preferred embodiments, the driving electrodes are patterned and arranged on the substrate at intervals, a detection area capable of accommodating the liquid drop to be detected is correspondingly arranged above the driving electrode of each interval part, the control part further controls the output module to output a corresponding voltage signal to the driving electrode to control the movement of the liquid drop, and when the liquid drop detection device is specifically applied, the movement, the separation and the combination of the liquid drop can be realized by electrifying different electrodes in the patterned electrodes based on a dielectric wetting principle.

In other preferred embodiments, the image processing module comprises a camera for acquiring the morphology of the droplets; and a main control device connected with the image pickup device and used for analyzing and processing the image data.

When the method is specifically applied, the image pickup device is used for acquiring the morphology of the liquid drops, and the main control device is used for analyzing and processing image data to detect the contact angle of the liquid drops with different sizes and shapes in real time.

A third aspect of the present invention provides a detection method using the detection device, the sensing member includes a top plate movably disposed with the substrate, and at least one sensing electrode disposed on the top plate, the top plate and the substrate being spaced apart to form the channel; the detection method at least comprises the following steps:

a. providing a drop to be measured into the channel;

b. after the top plate is driven to move to an initial position relative to the base plate by using a control device, detecting the position information of the sensing electrode by using a positioning device and feeding back the position information to the control part, wherein the control part further adjusts the position of the sensing electrode based on the position information to enable the end part of the sensing electrode to be in contact with the liquid drop to be detected, so that an electric connection path is formed between the sensing electrode and the driving electrode through the liquid drop to be detected;

c. And applying voltage to the driving electrode and the sensing component by using an output module, and acquiring a drop angle and a morphology change chart of the drop to be detected under corresponding conditions by using an image processing module.

The scheme provides a method for detecting drop angle and morphology change under sealed state, can avoid drop volatilization, can also be through the equal contact of the tip of controlling means automatic control sensing electrode and the not measuring drop of equidimension to make the sensing electrode form the electric connection passageway through measuring drop and driving electrode, be convenient for follow-up circular telegram detection.

In other preferred embodiments, at least two droplets to be measured having different sizes in step a are located in the channel, and step b further comprises: and respectively adjusting the distances between the sensing electrodes corresponding to different liquid drops to be detected one by one and the corresponding liquid drops to be detected.

In this embodiment, at least two droplets to be detected with different sizes can be detected simultaneously, and it is to be understood that the detection device and the detection method provided by the invention can be completely suitable for detecting more droplets with different sizes simultaneously and acquiring the morphology and the droplet angle of each droplet in real time under the condition that the control device is in one-to-one correspondence control.

The fourth aspect of the invention provides a sequencing device comprising the microfluidic chip, which adopts a sealed chip device, not only can prevent volatilization, but also can accurately control the upper electrode (the end part of the sensing electrode) to be contacted with liquid drops with different sizes, and the whole flow is configured automatically, so that the manual operation time is greatly reduced, and the accuracy of test data can be effectively improved.

The fifth aspect of the present invention provides a method for manufacturing a droplet control chip for a sequencer, the method comprising determining a process parameter of the droplet control chip by using the detection device, wherein the process parameter at least comprises position information and a voltage control parameter; the sensing component comprises a top plate which is movably arranged with the base plate and at least one sensing electrode arranged on the top plate, and the channel is formed between the top plate and the base plate at intervals; determining the process parameters of the liquid drop control chip comprises:

selecting a droplet to be detected with a specified size, and applying the droplet to be detected into the channel;

adjusting the relative distance between the top plate and the substrate, and further adjusting the position between the sensing electrode and the liquid drop to be detected, so as to obtain the position information corresponding to the top plate, the substrate, the sensing electrode and the driving electrode when the end part of the sensing electrode is in contact with the liquid drop to be detected;

The output module is regulated to output different voltage driving signals to the electric connection channel, and voltage control information corresponding to the liquid drop to be detected is obtained;

and determining the optimal voltage control parameters based on the morphology and the contact angle of the liquid drop to be detected under different voltage driving signals detected by the image processing module.

Compared with the prior art, the invention has the following beneficial effects:

according to the microfluidic chip provided by the first aspect of the invention, the shape and the size of the liquid drop to be detected can be sensed by using the sensing component, sensed information (such as the distance between the liquid drop to be detected and the sensing component) is fed back to the control component, and the control component adjusts the position of the sensing component up and down according to the sensed information, so that the distance between the sensing component (namely the upper electrode) and the liquid drop to be detected with different sizes is accurately controlled, and the accuracy of test data is improved; when the microfluidic chip adopts a sealing structure, the volatilization rate of liquid drops can be effectively slowed down;

the detection device comprising the microfluidic chip provided by the second aspect of the invention aims at the liquid drop to be detected, adjusts the position of the sensing electrode to enable the sensing electrode to be in contact with the liquid drop to be detected, provides different voltage driving signals for the electric connection path, and finally acquires the morphology of the liquid drop in the channel under the different voltage driving signals through the image processing module, and detects the contact angle of the liquid drop to be detected;

The third aspect of the invention provides a method for detecting the angle and morphology change of liquid drops in a sealing state, which can avoid volatilization of the liquid drops, and can automatically control the end part of a sensing electrode to be contacted with liquid drops to be detected with different sizes through a control device so that the sensing electrode forms an electric connection path with a driving electrode through the liquid drops to be detected, and the method is automatic in control, convenient and quick and high in accuracy;

the fourth aspect of the present invention provides a sequencing device comprising the microfluidic chip, wherein the sequencing device is a sealed chip device, which can prevent volatilization and accurately control the contact of an upper electrode (the end part of a sensing electrode) with droplets of different sizes;

the fifth aspect of the present invention provides a method for manufacturing a droplet control chip for a sequencer, which uses the microfluidic chip with controllable distance between a substrate and a top plate, can quickly and conveniently obtain multiple process parameters adapted to a target droplet, accurately prepares a corresponding droplet control chip by combining the obtained process parameters, is suitable for quick detection of a multi-channel and complex system, quickly locates and optimizes the process parameters and conditions, and saves process fumbling time and cost.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.

Fig. 1 is a schematic structural diagram of a microfluidic chip according to one embodiment of the present invention;

FIG. 2 is a schematic structural diagram of a detection device according to one embodiment of the present invention;

FIG. 3 is a schematic structural diagram of a detecting device according to another embodiment of the present invention;

FIG. 4 is a schematic diagram of a droplet to be measured on a driving electrode according to the present invention;

FIG. 5 is a second diagram of a droplet to be measured on a driving electrode according to the present invention;

FIG. 6 is a schematic diagram of the state of the droplet in a detection process of the prior art device;

FIG. 7 is a schematic diagram of the sequencing device according to the present invention showing the state of the droplet at the completion of detection;

FIG. 8 is a graph showing the trend of the change of the water drop angle of EB liquid with voltage detected by the prior device;

FIG. 9 is a graph showing the trend of the detected water drop angle of ultrapure water with voltage in the prior art;

FIG. 10 is a graph showing the trend of the change of the drop angle of the ultra-pure water and EB liquid with voltage detected by the sequencing device;

FIG. 11 is a real-time feedback diagram of voltage parameters according to an embodiment of the invention.

Reference numerals

101. A substrate; 102. a driving motor; 103. a temperature control component;

200. a sensing member; 201. a top plate; 202. a sensing electrode;

300. a sidewall; 301. an inner wall; 302. an outer wall;

400. A driving electrode;

500. a drop to be measured;

600. a light source;

700. an output module;

800. an image processing module; 801. an image pickup device; 802. and a master control device.

Detailed Description

To further clarify the above and other features and advantages of the present invention, a further description of the invention will be rendered by reference to the appended drawings. It should be understood that the specific embodiments presented herein are for purposes of explanation to those skilled in the art and are intended to be illustrative only and not limiting.

In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential", etc. indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings are merely for convenience in describing the present invention and simplifying the description, and do not indicate or imply that the device or element being referred to must have a specific orientation, be configured and operated in a specific orientation, and therefore should not be construed as limiting the present invention.

Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include at least one such feature. In the description of the present invention, the meaning of "plurality" means at least two, for example, two, three, etc., unless specifically defined otherwise.

In the present invention, unless explicitly specified and limited otherwise, the terms "mounted," "connected," "secured," and the like are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; either directly or indirectly, through intermediaries, or both, may be in communication with each other or in interaction with each other, unless expressly defined otherwise. The specific meaning of the above terms in the present invention can be understood by those of ordinary skill in the art according to the specific circumstances.

In the present invention, unless expressly stated or limited otherwise, a first feature "up" or "down" a second feature may be the first and second features in direct contact, or the first and second features in indirect contact via an intervening medium. Moreover, a first feature being "above," "over" and "on" a second feature may be a first feature being directly above or obliquely above the second feature, or simply indicating that the first feature is level higher than the second feature. The first feature being "under", "below" and "beneath" the second feature may be the first feature being directly under or obliquely below the second feature, or simply indicating that the first feature is less level than the second feature.

In the description of the present specification, a description referring to terms "one embodiment," "some embodiments," "examples," "specific examples," or "some examples," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, the different embodiments or examples described in this specification and the features of the different embodiments or examples may be combined and combined by those skilled in the art without contradiction.

As shown in fig. 1, the present invention provides a microfluidic chip in a first aspect, where the microfluidic chip at least includes a substrate 101 and a sensing device that are disposed opposite to each other, a driving electrode 400 is disposed on the substrate 101, the driving electrode 400 is disposed on a side of the substrate 101 opposite to the sensing device, and a droplet can be dropped above the driving electrode 400; channels for accommodating the droplets 500 to be measured are formed between the substrate 101 and the sensing device, the number and the shape of the channels are not limited, and a plurality of droplets 500 to be measured with different shapes and sizes can be accommodated; the sensing device comprises a control component and a sensing component 200 which are connected with each other, wherein the control component can control at least one of the sensing component 200 and the substrate 101 to move relatively towards a direction approaching or separating from each other, so that the sensing component 200 forms an electric connection path with the driving electrode 400 through the liquid drop 500 to be detected, and further detection of the drop angle of the liquid drop 500 to be detected is realized.

In the specific implementation process, the to-be-detected liquid drops 500 with different shapes and components can be dripped into the channel above the driving electrode 400, and the drop angles of a plurality of liquid drops with different shapes can be detected simultaneously; the control part can control at least one of the sensing part 200 and the substrate 101 to move relatively in a direction approaching or separating from each other, namely, can control the sensing part 200 to move or control the substrate 101 to move, and can also control both to move, so that the tail end of the sensing part 200 can be contacted with liquid drops with different sizes and shapes, the water drop angle can be accurately detected conveniently, the sensing part 200 can sense the shape and the size of the liquid drop 500 to be tested, the sensed distance between the liquid drop 500 to be tested and the sensing part 200 is fed back to the control part, and the control part adjusts the position of the sensing part 200 (or the position of the substrate 101) up and down according to the distance between the sensing part 200 and the liquid drop 500 to be tested with different sizes, so that the accuracy of test data is improved.

In this embodiment, the sensing component 200 can sense a plurality of droplets 500 to be detected with different shapes and sizes at the same time, and only the different portions of the sensing component 200 are required to be controlled to be in one-to-one correspondence with the droplets 500 to be detected for detection, so that the application range is wide.

Further, the microfluidic chip further includes a sidewall 300 connected to the top plate 201 and extending toward the substrate 101; the sensing part 200 includes a top plate 201 and at least one sensing electrode 202 (only one sensing electrode 202 protruding therefrom is shown in fig. 1) provided on the top plate 201; the side wall 300, the top plate 201 and the substrate 101 enclose a sealed cavity, the drop 500 to be tested is positioned in the sealed cavity, so that drop volatilization can be effectively avoided, and when the control component controls the top plate 201 to drive the sensing electrode 202 to move, the top plate 201 changes in position relative to the substrate 101 in the height direction of the side wall 300, namely the top plate 201 moves up and down; the drive electrode 400 is at least partially located within the sealed cavity.

In specific use, the side wall 300, the top plate 201 and the substrate 101 enclose a sealed cavity, the shape of the side wall 300 is not limited herein, the side wall 300 may be a circular side wall 300, or may be a side wall 300 formed by a plurality of planes, the droplet 500 to be tested is in a sealed environment, the problem of volatilization of the droplet can be effectively solved, at least one sensing electrode 202, such as a lead screw electrode, is disposed on the top plate 201, the tip of the electrode can be in contact with the top of the droplet 500 to be tested, and can be used for detecting signals of the droplet in the process of controlling the droplet by the microfluidic chip, the number of the sensing electrodes 202 is in one-to-one correspondence with the number and positions of the droplet 500 to be tested, when the control component controls the top plate 201 to drive the sensing electrode 202 to move, the top plate 201 changes up and down in the height direction of the side wall 300 relative to the substrate 101, the vertical distance between the sensing electrode 202 and the droplet 500 to be tested can be adjusted, the driving electrode 400 is at least partially located in the sealed cavity, the upper surface area of the substrate 101 is larger than the bottom area of the sealed cavity, so that the droplet 500 to be completely sealed, the droplet 500 to be placed above the portion of the driving electrode 400 located in the sealed cavity, the sensing electrode 202 can be electrically connected with the droplet 500 to be electrically detected by the driving electrode 400, and the droplet 500 to be detected, thereby realizing the detection of the passage 500.

Optionally, a liquid adding hole is reserved on the top plate 201, and the liquid drop 500 to be measured can be dripped into the channel through the liquid adding hole, so that the top plate 201 does not need to be taken out, and the operation is convenient.

Further, at least one driving electrode 400 is disposed on the upper surface of the substrate 101, at least one detection area is disposed on the upper surface of the driving electrode 400, at least one droplet 500 to be detected can be dropped in the detection area, and the sensing electrode 202 corresponds to the droplets 500 to be detected in the detection area below the detection area one by one; the sensing electrodes 202 are arranged in one-to-one correspondence with the detection areas, the sensing electrodes 202 are generally located above the detection areas corresponding to the sensing electrodes, the distance between the sensing electrodes 202 and the liquid drops 500 to be detected can be adjusted through up-down movement of the sensing electrodes 202, and the microfluidic chip further comprises a positioning device for positioning one side, close to the detection areas, of the sensing electrodes 202, the distance between the sensing electrodes 202 and the liquid drops can be monitored through the positioning device, and the distance between the sensing electrodes 202 and the liquid drops 500 to be detected with different sizes can be adjusted conveniently.

In other embodiments, PDMS, glass, PC, PMMA, etc. may be used as the material of the substrate 101, and the upper surface of the substrate 101 is provided with a plurality of driving electrodes 400, and the driving electrodes 400 may be designed into different shapes, such as different patterns of square, rectangle, crescent, etc., or may be designed into different pitches, such as equal pitches, unequal pitches, gradient pitches, etc., and the driving electrodes 400 are led out to the outside through wirings inside the substrate 101 and connected to external control electrodes.

Further, all the sensing electrodes 202 are movably disposed on the top plate 201, and are connected to the control part; the control part further controls the sensing electrode 202 to move toward or away from the droplet 500 to be measured relative to the top plate 201 on the basis of the movement of the top plate 201 relative to the substrate 101.

In a specific embodiment, in an initial state, all the sensing electrodes 202 are located inside the top plate 201, after a drop 500 to be detected is dripped into a certain detection area, the control component can control the sensing electrodes 202 at positions corresponding to the drop 500 to be detected to extend, and then the control component further controls the sensing electrodes 202 to move towards a direction approaching or separating from the drop 500 to be detected relative to the top plate 201 after the top plate 201 reaches an initial determined position on the basis of the movement of the top plate 201 relative to the substrate 101, so that the tip of the sensing electrode 202 contacts with the drop 500 to be detected, and signal detection is facilitated for the drop 500 to be detected.

In other embodiments, the movement of the substrate 101 can be controlled, or the substrate 101 and the top plate 201 can be controlled to move simultaneously, to adjust the spacing between the sensing electrode 202 and the droplet 500 to be measured, so that the top of the droplet 500 to be measured is in contact with the tip of the sensing electrode 202.

Further, the side wall 300 is provided with a light transmitting portion for light to pass through, and the light transmittance of the light transmitting portion is greater than or equal to 0.8, so as to ensure a better light transmitting effect, the external light source 600 is used for making light incident into the sealed cavity through the light transmitting portion, and the light transmitting portion is highly transparent, so that light penetration and image acquisition are facilitated.

In this embodiment, the material of the transparent portion is not specifically limited, but the transmittance of the transparent portion is required to be greater than or equal to 0.8 to ensure a better transparent effect, and the number, shape and size of the transparent portion are not specifically limited, for example, one or more circular transparent sheets may be used, or one side of the entire sidewall 300 may be used as the transparent portion.

Further, the side wall 300 includes an inner wall 301 made of a transparent material and an outer wall 302 made of an opaque material, and the light-transmitting portion is disposed on the outer wall 302.

The inner wall 301 and the outer wall 302 in this scheme can be directly attached to each other as shown in fig. 1, and a gap can be left between them, a transparent part is disposed on the opaque outer wall 302, the transparent part is still in a sealed state, the external light source 600 is aligned to the transparent part, the light source 600 irradiates the light into the sealed cavity through the transparent part and the transparent inner wall 301, and irradiates the liquid drop to be measured, so that the subsequent image acquisition work can be performed normally. In other embodiments. The outer wall 302 may be integrally provided as a light-transmitting portion on the side opposite to the light source 600.

Further, the sensing electrode 202 includes a wire electrode and a shaft sleeve axially sleeved on the periphery of the wire electrode, the shaft sleeve drives the wire electrode to be movably connected with the top plate 201, and the shaft sleeve can effectively protect the wire electrode embedded therein, so that the sensing electrode can normally and stably perform a sensing function.

The electrode wire may be made of copper, gold, carbon fiber, etc. and has a diameter of 10-500 microns.

In another embodiment, the electrode wire is a metal wire with a diameter of 10-500 μm, a diameter of 1-10mm of the shaft sleeve, the metal wire is fixed by a fixing glue and embedded in the middle of the shaft sleeve, the fixing glue can be epoxy resin, acrylic ester or polyurethane, the molecular weight is 2 ten thousand-20 ten thousand, the position of the metal wire relative to the droplet 500 to be detected on the substrate 101 is adjusted by moving the shaft sleeve up and down, and the position accuracy of the metal wire can be adjusted by optical CMOS.

In other preferred embodiments, the driving electrode 400 is a patterned electrode sheet, and a first protective layer disposed on a side of the electrode sheet adjacent to the top plate 201; the first protective layer includes at least a first dielectric layer and a first hydrophobic layer, and the driving electrode 400 is used to control movement of the droplet. The upper surface of the electrode plate is required to be provided with a dielectric layer and a hydrophobic layer, or the two-in-one dielectric hydrophobic layer can be realized by means of film pasting, spraying, deposition and the like, the dielectric layer is used for maintaining the insulativity between the circuit and each layer, and different dielectric layer thicknesses can be designed according to different design requirements.

The liquid drop in the microfluidic chip in the scheme can be carried out in a water-in-oil or oil-in-water mode, and mainly depends on the property of the test liquid drop, so that the problem that the existing water drop angle test technology can only detect the contact angle of water or water dispersion systems (including solutions) and organic solvent systems in air can be solved.

In another preferred embodiment, the top plate 201 is provided with a second protective layer on a side close to the substrate 101; the second protective layer comprises at least a conductive layer and a second hydrophobic layer located on a side of the conductive layer near the substrate 101. The material of the top plate 201 may be PDMS, glass, PC, PMMA, etc., and the lower surface of the top plate 201 may be required to be formed with a conductive layer by spraying, depositing, sputtering, etc., or a conductive material may be used, and the conductive layer may be required to be formed with a hydrophobic layer, or a conductive and hydrophobic material may be used.

In other preferred embodiments, the microfluidic chip further includes a temperature control component 103 connected to the substrate 101, where the temperature control component 103 is configured to adjust the temperature of the substrate 101 to control the temperature in the sealed cavity to 4-97 ℃, so that the evaporation problem existing in the existing droplet corner testing device can be solved, and the evaluation and detection of the droplet in the heating state and the PCR process can be performed.

In another embodiment, the temperature control part 103 is composed of metal electrodes and sensors, and mainly controls the temperature inside the chip, so as to provide different temperature time curve control and different temperature rise and fall rate control.

As shown in fig. 1, in other embodiments, an outer wall 302 and an inner wall 301 of the microfluidic chip are attached, the outer wall 302 is made of epoxy resin, the inner wall 301 is made of elastic materials such as silicone rubber, polydimethylsiloxane, or the like, the top plate 201 is made of one or more of PDMS, PMMA, quartz glass, or the like, a patterned electrode is disposed on the substrate 101, the area of the patterned electrode is smaller than the upper surface area of the substrate 101, a droplet 500 to be detected is dropped into a channel above the patterned electrode, the droplet 500 to be detected contains components such as surfactant, biochemical enzyme reagent, or the like, the substrate 101 is made of one or more of PC, glass, PCB board, or the like, fine electrode wires are vertically disposed on the lower surface of the top plate 201, and a CMOS sensor capable of precisely positioning the electrode wire is mounted on the outer wall 302.

In other embodiments, the material of the microfluidic chip is glass or plastic, and layout design can be realized based on the digital microfluidic principle to complete merging and splitting of droplets, as shown in fig. 4, the driving electrodes 400 are arranged in an array, the single driving electrode 400 may be square and distributed on the substrate in a grid shape as a whole, two droplets 500 to be detected are respectively located in two detection areas with numbers of 6 and 7, and the two droplets 500 to be detected can be guided to move, split and merge between different areas by adjusting the potentials of different detection areas.

As shown in fig. 5, the droplets 500 to be measured with different sizes are respectively located in the detection areas of numbers 6-7 and 14-15, and the movement, splitting and merging of the droplets 500 to be measured with different sizes can be realized by respectively adjusting the potentials of the detection sites.

Further, the second aspect of the present invention provides a detection device comprising a microfluidic chip, the detection device further comprising a light source 600; and

an output module 700 including a first output terminal connected to the driving electrode 400 and a second output terminal connected to the sensing part 200; the output module 700 is capable of outputting different voltage driving signals to the electrical connection path; the voltage driving signals output by the voltage output module 700 may have different waveforms and frequencies;

the image processing module 800 is configured to collect the morphology of the droplet in the channel under different voltage driving signals, and detect the contact angle of the droplet.

As shown in fig. 3, when the detection device is specifically applied to detect the contact angle of the droplet 500 to be detected, the LED light source 600 is first aligned to the light-transmitting portion, so that light enters the cavity and is aligned to the droplet 500 to be detected, the position of the sensing electrode 202 is adjusted to make contact with the droplet 500 to be detected, the output module 700 is opened, the driving electrode 400 and the sensing electrode 202 are pressurized, different voltage driving signals are provided to the electrical connection path, finally the image processing module 800 is used to collect the morphology of the droplet in the path under the different voltage driving signals, and detect the contact angle of the droplet 500 to be detected, and the whole process is automatically performed under the sealed environment, so as to reduce the volatilization of the droplet.

It should be added that the main structure of the microfluidic chip in this embodiment is made of epoxy resin, rubber, glass, etc.

Further, the driving electrodes 400 are patterned and arranged on the substrate 101 at intervals, and can be designed into different shapes, such as different patterns of square, rectangle, crescent and the like, and different pitches, such as equal pitch, unequal pitch, gradient pitch and the like, a detection area capable of containing the droplet 500 to be detected is correspondingly arranged above each driving electrode 400 of each interval part, and the control part further controls the output module 700 to output a corresponding voltage signal to the driving electrodes 400 to control the movement of the droplet.

In a specific operation, when a liquid exists on the driving electrode 400 and an electric potential is applied to the driving electrode 400 by using the electrowetting principle, the wettability of the solid-liquid interface at the corresponding position of the driving electrode 400 can be changed, the contact angle between the liquid drop and the interface of the driving electrode 400 is changed, and if the electric potential difference exists between the electrodes in the liquid drop area, a transverse driving force is generated when the contact angle is different, so that the liquid drop transversely moves on the electrode substrate 101, and different electrodes in the patterned electrode are electrified to realize movement, separation and combination of the liquid drop.

In other preferred embodiments, the image processing module 800 includes a camera 801 for acquiring the morphology of the droplets; and a main control device 802 connected to the image pickup device 801 and configured to analyze and process image data.

In a specific application, the image capturing device 801 (such as a CMOS camera) is used to obtain the morphology of the liquid drops, and the image data is analyzed and processed by the main control device 802 to detect the contact angle of the liquid drops with different sizes and shapes in real time.

The image processing module 800 can photograph the morphology of the liquid drop, then automatically or manually take points to measure the contact angle, output contact angle data, record and save video, record the change processes of movement, separation, mixing and the like of the liquid drop, and the like.

In a specific embodiment, as shown in fig. 2, the detection device is used to detect the drop angle and the shape change of the drop in the open environment, the light source 600 is LED light, the bottom substrate 101 is one or more of glass, PC or PCB, the PI film is coated on the light source, the PI film is divided into patterned electrodes, the output module 700 is a programmable voltmeter, which can generate an ac/dc voltage with adjustable frequency of-500V to +500V, the CMOS camera and the main control computer are arranged on the same side of the drop, the LED light source 600 is arranged on the other side of the drop, the first output end of the programmable voltmeter is connected with an electrode positive electrode, which applies a voltage to a corresponding electrode through a bonding pad on the biological slide, the second output end is connected with an electrode negative electrode, which is a metal wire material conducting, the diameter is 10 um-500 um, and the detection device can complete functions of power supply, test program as required, automatic recording and feedback of the programmed program as shown in the figure.

In a specific embodiment, as shown in fig. 3, the detection device is used to detect the angles and morphology changes of a plurality of droplets in a closed environment, in which the driving motor 102 (control component) is only schematically represented, the structure and the connection relation with other structures are not represented, at least two control interfaces are provided on the driving motor 102 (control component), one control interface is used to control the electrode wire corresponding to the position of the droplet 500 to be detected to extend out of the top plate 201, the other control interface is used to drive the top plate 201 to make the electrode wire connected thereto contact with the droplet or leave the droplet, the CMOS camera is disposed obliquely above one side of the microfluidic chip, and the LED light source 600 is respectively located at two sides of the droplet 500 to be detected, the positions of the electrode wire are calibrated and adjusted by using CMOS, the material used for the top plate 201 is one or more of PDMS, PMMA, quartz glass, etc., the electrode negative electrode wire (i.e., the electrode wire) is a conductive metal wire, the diameter is 10 um-500 um, the substrate 101 is one or more of glass, PC or PCB, the upper is coated with PI film, and is divided into patterns, the CMOS camera is disposed above the electrode wire, and the electrode wire is heated in the range of the cooling chamber(s) is heated by the cooling semiconductor 101) and the cooling chamber(s) is set in the cooling chamber(s) 4-97).

Further, a third aspect of the present invention provides a detection method using the above detection device, where the sensing component 200 includes a top plate 201 movably disposed with the substrate 101, and at least one sensing electrode 202 disposed on the top plate 201, and a channel is formed between the top plate 201 and the substrate 101 at intervals, and the channel can accommodate a droplet 500 to be detected; the detection method at least comprises the following steps:

a. providing a drop 500 to be tested in the channel, and dripping the drop 500 to be tested at a liquid adding hole on the top plate 201;

b. after the control device is used for driving the top plate 201 to move to an initial position relative to the substrate 101, the positioning device is used for detecting the position information of the sensing electrode 202 and feeding back the position information to the control component, the control component further adjusts the position of the sensing electrode 202 based on the position information, so that the end part of the sensing electrode 202 is in contact with the liquid drop 500 to be detected, the sensing electrode 202 forms an electric connection path with the driving electrode 400 through the liquid drop 500 to be detected, and in the initial position, the distance between the top 201 and the liquid drop 500 to be detected is not more than 500 mu m, so that the micro-fluidic chip is convenient for controlling the movement of the liquid drop 500 to be detected, meanwhile, the liquid drop 500 to be detected can contact with the tail end of the sensing electrode 202, and meanwhile, the influence of roughness on the surface of the top plate 201 on the liquid drop 500 is avoided;

c. The output module 700 is used for applying voltage to the driving electrode 400 and the sensing component 200, and the image processing module 800 is used for acquiring a drop angle and a morphology change chart of the drop 500 to be detected under corresponding conditions, so that the contact angle of the drop can be detected, videos can be recorded and stored, and the change processes of movement, separation, uniform mixing and the like of the drop can be recorded.

The present solution provides a method for detecting drop angle and morphology change in a sealed state, which can avoid drop volatilization, and can automatically control the end of the sensing electrode 202 to contact with the drop 500 to be detected with different sizes through the control device, so that the sensing electrode 202 forms an electrical connection path with the driving electrode 400 through the drop 500 to be detected, so that subsequent power-on detection is facilitated, and the voltage generated by the voltage generator (i.e. the output module 700) in the embodiment is time-synchronized with the monitoring picture, so that the image and the voltage waveform can be corresponding.

It should be noted that, the "initial position" in this embodiment refers to an operation of adjusting the distance between the top plate 201 and the droplet to be measured, after the distance between the sensing electrode 202 and the driving electrode 400 is within 2000 μm after coarse adjustment, detecting the position information of the sensing electrode 202 by the positioning device and feeding back to the control unit, and the control unit further adjusts the position of the sensing electrode 202 based on the position information, so that the end of the sensing electrode 202 is in contact with the droplet 500 to be measured, i.e., "fine adjustment", which is similar to the operation when the microscope is used to observe the specimen.

Further, in step a, at least two droplets 500 to be measured having different sizes are located in the channel, and step b further includes: the distances between the sensing electrodes 202 corresponding to different droplets 500 to be measured and the corresponding droplets 500 to be measured are respectively adjusted.

In this embodiment, at least two droplets 500 of different sizes can be detected simultaneously, and it is to be understood that the detection device and the detection method provided by the invention can be completely suitable for detecting more droplets of different sizes simultaneously and acquiring the morphology and the droplet angle of each droplet in real time under the condition that the control device is in one-to-one correspondence control.

The droplet 500 to be measured in the present invention may be an aqueous system (pure water, surfactant solution, aqueous solution of enzyme preparation with organic solvent), oil-water system (droplet on silicone oil), etc., and contains surfactant, biochemical enzyme reagent, etc.

Further, the fourth aspect of the present invention provides a sequencing device including the microfluidic chip, and the sealing chip device is adopted, so that not only can volatilization be prevented, but also the upper electrode (the end of the sensing electrode 202) can be accurately controlled to be contacted with liquid drops with different sizes, the whole flow is automatically configured, the manual operation time is greatly reduced, and the accuracy of test data can be effectively improved.

Further, a fifth aspect of the present invention provides a method for manufacturing a droplet control chip for a sequencer, the method comprising determining a process parameter of the droplet control chip by using the detection device, the process parameter at least comprising position information and a voltage control parameter; the sensing part 200 includes a top plate 201 movably disposed with the substrate 101 and at least one sensing electrode 202 disposed on the top plate 201, and a channel is formed between the top plate 201 and the substrate 101 at an interval; determining the process parameters of the droplet control chip includes:

selecting a droplet 500 to be measured with a specified size, and applying the droplet 500 to be measured into the channel;

adjusting the relative distance between the top plate 201 and the substrate 101, and further adjusting the position between the sensing electrode 202 and the droplet 500 to be detected, so as to obtain position information corresponding to the top plate 201, the substrate 101, the sensing electrode 202 and the driving electrode 400 when the end part of the sensing electrode 202 is in contact with the droplet 500 to be detected;

the adjusting output module 700 outputs different voltage driving signals to an electric connection path comprising the sensing electrode 202, the liquid drop 500 to be detected and the driving electrode 400, and acquires voltage control information corresponding to the liquid drop 500 to be detected;

the optimum voltage control parameters are determined based on the morphology and the contact angle of the droplet 500 to be measured under different voltage driving signals detected by the image processing module 800, and the signals acquired by the droplet 500 to be measured are detected by the sensing electrode 202.

It should be noted that, in this embodiment, the substrate used in the microfluidic chip is at least one of PDMS, glass, PC and PMMA, the thickness of the first dielectric layer is 0-2000 nm, the types of the first dielectric layer are SiN, alumina, PTFE, parylene and PDMS, the distance between the sensing electrode 202 and the driving electrode 400 can be adjusted to be within 2000 μm by adjusting the distance between the top plate 201 and the substrate 101, the distance between the sensing electrode 202 and the droplet 500 to be measured can be further adjusted in combination with the position of the sensing electrode 202, the distance between the sensing electrode 202 and the driving electrode 400 can be finely adjusted to be within 500 μm, and the optimal process conditions and parameters can be determined by one process measurement, wherein the respective parameter variables can be continuous or random.

In the preparation process of the microfluidic chip, the voltage measurement module, the morphology image measurement module and the like are integrated, the sensing electrode 202 can detect the threshold voltage, the saturation voltage, the breakdown voltage and the like under the unipolar plate or the bipolar plate, the image processing module 800 can detect the variation quantity and the variation rate of the contact angle of the liquid drop, the photoelectric probe can also be used for detecting the movement rate of the liquid drop and the like, the optimal power supply voltage condition and the parameters can be determined through one flow, the chip with various specifications does not need to be customized for carrying out small-batch verification, the chip design volume is small, the loss and the later recovery processing pollution to the biochemical reagent are small, the process time and the production cost are saved, and the microfluidic chip is worth popularizing.

The following describes the detection of the change in drop angle versus topography of drops under different conditions in detail in connection with the examples.

In one embodiment, as shown in fig. 1, a spotter or a sample injection needle is used to drop 2 microliters of liquid drop on the driving electrode 400, the top plate 201 is immediately covered, the position of the top plate 201 is adjusted, the electrode wire end on the lower surface of the top plate 201 reaches the instrument cursor indication area, and the positioning device is a CMOS sensor mounted on the side wall of the chip, and the CMOS sensor senses the position of the electrode wire. The output module 700 is an oscilloscope and a power supply generating device, the output module 700 applies positive and negative 10V, 20V, 50V, 100V, 150V, 200V, 300V and 400V to an electrical connection path where the driving electrode 400 is located, the pulse waveform is one of square wave, triangular wave and sine wave, the pulse time is 0.5 seconds, 1 second, 1.5 seconds and 2 seconds, and the duty ratio is 1: 1.

The water drop angle and morphology change diagrams under different conditions are tested, the voltage range of the power supply generating device is plus or minus 800V, the contact angle range is 0-130 degrees, and the movement rate is converted by adopting a plurality of photoelectric detection probes through software, so that the description is omitted.

In another embodiment, as shown in fig. 1, a spotter or a sample injection needle is used to drop 5 microliters of liquid drop on the driving electrode 400, the top plate 201 is immediately covered, and the position of the top plate 201 is adjusted so that the end of the electrode wire on the lower surface of the top plate 201 reaches the instrument cursor indication region. The temperature control device is adjusted to enable the temperature in the sealing cavity to be changed within 95 degrees, 60 degrees and 72 degrees, and the switching time is 15 seconds;

Using an oscilloscope and a power supply generating device, applying positive and negative 10V, 20V, 50V, 100V, 150V, 200V, 300V and 400V, wherein the pulse waveform is one of square wave, triangular wave and sine wave, the pulse time is 0.5 seconds, 1 second, 1.5 seconds and 2 seconds, and the duty ratio is 1: 1.

The change patterns of the water drop angle and the morphology under different conditions are tested, the voltage range of the power supply generating device is plus or minus 800V, the contact angle range is 0-130 degrees, and the movement rate is converted by adopting a plurality of photoelectric detection probes through software.

In another embodiment, as shown in fig. 1, 5 droplets of 2 microliters are dripped on the driving electrode 400 by using a spotter or a sample injection needle, the top plate 201 is immediately covered, and the position of the top plate 201 is adjusted so that the electrode wire end on the lower surface of the top plate 201 reaches the instrument cursor indication region. Using an oscilloscope and a power supply generating device, applying positive and negative 10V, 20V, 50V, 100V, 150V, 200V, 300V and 400V, wherein the pulse waveform is one of square wave, triangular wave and sine wave, the pulse time is 0.5 seconds, 1 second, 1.5 seconds and 2 seconds, and the duty ratio is 1: 1.

Meanwhile, a plurality of water drop angles and morphology change diagrams under different conditions are tested, the voltage range of a power supply device is positive and negative 800V, the contact angle range is 0-130 degrees, and the movement rate is converted by adopting a plurality of photoelectric detection probes through software.

In another embodiment, as shown in fig. 1, 5 droplets of 5 microliters are dripped on the driving electrode 400 by using a spotter or a sample injection needle, the top plate 201 is immediately covered, and the position of the top plate 201 is adjusted so that the electrode wire end on the lower surface of the top plate 201 reaches the instrument cursor indication region. The temperature control device is regulated to change the temperature in the sealing cavity within 95 degrees, 60 degrees and 72 degrees, and the switching time is 15 seconds;

As shown in the following table 1, a plurality of different conventional test conditions and water drop angles under specific test conditions were detected by using the detection device in the prior art and the sequencing device provided by the present application, respectively, and test performance statistics under different test conditions are shown in table 1.

As can be seen from Table 1, the sequencing device provided by the application is superior to the detection device in the prior art in terms of at least the precision of electrode adjustment, time consumption in the detection process, simultaneous detection of a plurality of liquid drops and the like, can also effectively avoid volatilization of liquid drops, can detect the drop angle of the liquid drops in a heating state, and has the functions of automatic detection, automatic data arrangement and automatic equipment protection, and is downward compatible with a plurality of database builder chips and sequencing chips.

Table 1 presents a list of comparison of the performance of the apparatus and the drop angle test of the sequencing device of the present application

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In addition, as shown in fig. 6, when the detection device in the prior art is used to detect the drop angle, the shape and size of the drop will change due to volatilization during the detection process; as shown in fig. 7, when the sequencing device of the present application is used to detect the drop angle, the drop remains in its original state until the detection is completed, and the form and size of the drop do not change during the whole detection process.

When the existing device is used for detecting the water drop angle of the EB liquid, as the EB liquid contains components such as a surfactant, an additive and the like, the size of the liquid drop changes rapidly (for example, from 3-5 microliters to 2-3 microliters) along with the volatilization of water, the concentration of each component changes greatly, the influence on the performance such as the water drop angle, rheological property and the like is great, and as shown in figure 8, the fluctuation of data is great during the process test. As shown in fig. 9, when the conventional apparatus is used to detect the drop angle of ultrapure water (four sets of data are tested), the drop volatilization causes large data fluctuation during the test.

As shown in FIG. 10, when the sequencing device provided by the invention is used for detecting the ultra-pure water drop angle and the EB liquid drop angle, the drop angle and the voltage are almost in a linear relation, and the fluctuation of data is small. The voltage in fig. 11 is a real-time voltage value at the droplet monitored by an oscilloscope when ±300V voltage is applied to the electrical connection path where the driving electrode 400 of the present invention is located.

The microfluidic chip provided by the first aspect of the present invention can precisely control the distance between the sensing component 200 and the droplets 500 to be tested with different sizes, thereby improving the accuracy of test data; when the microfluidic chip adopts a sealing structure, the volatilization rate of liquid drops can be effectively slowed down;

the detection device comprising the microfluidic chip provided by the second aspect of the present invention can automatically adjust the position of the sensing electrode 202 to make contact with the droplet 500 to be detected, then provide different voltage driving signals to the electrical connection path, and finally collect the morphology of the droplet in the channel under the different voltage driving signals through the image processing module 800, and detect the contact angle of the droplet 500 to be detected, so that the full-flow automatic configuration greatly reduces the operation time and improves the accuracy of the test data;

The third aspect of the present invention provides a method for detecting a drop angle and a morphology change of a drop in a sealed state, which can not only avoid drop volatilization, but also automatically control the end of a sensing electrode 202 to contact with drops 500 to be detected with different sizes through a control device, so that the sensing electrode 202 forms an electrical connection path with a driving electrode 400 through the drops 500 to be detected, and the method is convenient and fast to control automatically, and has high accuracy;

the fourth aspect of the present invention provides a sequencing device comprising a microfluidic chip, wherein the sequencing device is a sealed chip device, which can prevent volatilization and accurately control the contact of an upper electrode (the end part of a sensing electrode 202) with droplets with different sizes;

the fifth aspect of the invention provides a method for manufacturing a droplet control chip for a sequencer, which can automatically integrate various process parameters into the droplet control chip, is suitable for rapid detection of a multichannel and complex system, rapidly locates and optimizes the process parameters and conditions, and saves process fumbling time and cost.

While embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and not to be construed as limiting the invention, and that variations, modifications, alternatives and variations may be made to the above embodiments by one of ordinary skill in the art within the scope of the invention.

Claims

1. The microfluidic chip is characterized by at least comprising a substrate (101) and a sensing device which are oppositely arranged, wherein a driving electrode (400) is arranged on the substrate (101), and a channel for accommodating liquid drops (500) to be detected is formed between the substrate (101) and the sensing device;

the sensing device comprises a control component and a sensing component (200) which are connected with each other, wherein the control component can control at least one of the sensing component (200) and the substrate (101) to move relatively towards a direction approaching or separating from each other, so that the sensing component forms an electric connection path with the driving electrode (400) through the liquid drop (500) to be detected.

2. The microfluidic chip according to claim 1, wherein,

the sensing component (200) comprises a top plate (201) and at least one sensing electrode (202) arranged on the top plate (201);

the microfluidic chip further comprises a side wall (300) connected with the top plate (201) and extending towards the substrate (101); the side wall (300), the top plate (201) and the base plate (101) form a sealed cavity, and the control part controls the top plate (201) to change the position of the top plate (201) relative to the base plate (101) in the height direction of the side wall (300) when the top plate (201) drives the sensing electrode (202) to move; the drive electrode (400) is at least partially located within the sealed cavity.

3. The microfluidic chip according to claim 2, wherein the upper surface of the substrate (101) is provided with at least one of the drive electrodes (400), and the upper surface of the drive electrode (400) is provided with at least one detection region;

the sensing electrodes (202) are arranged in one-to-one correspondence with the detection areas, and the microfluidic chip further comprises a positioning device for positioning one side, close to the detection areas, of the sensing electrodes (202).

4. A microfluidic chip according to claim 3, wherein all the sensing electrodes (202) are movably arranged on the top plate (201) and are connected to the control means; the control part further controls the sensing electrode (202) to move towards or away from the liquid drop (500) to be detected relative to the top plate (201) on the basis of the movement of the top plate (201) relative to the base plate (101).

5. A microfluidic chip according to claim 3, wherein a light-transmitting portion through which light passes is provided on the side wall (300), and the light transmittance of the light-transmitting portion is greater than or equal to 0.8, and an external light source enters the sealed cavity through the light-transmitting portion.

6. The microfluidic chip according to claim 5, wherein the side wall (300) comprises an inner wall (301) of transparent material and an outer wall (302) of opaque material, the light-transmitting portion being provided on the outer wall (302).

7. The microfluidic chip according to any one of claims 3 to 6, wherein the sensing electrode (202) comprises a wire electrode and a shaft sleeve axially sleeved on the periphery of the wire electrode, and the shaft sleeve drives the wire electrode to be movably connected with the top plate (201).

8. The microfluidic chip according to claim 6, wherein the driving electrode (400) comprises a patterned electrode sheet and a first protective layer disposed on a side of the electrode sheet close to the top plate (201); the first protective layer comprises at least a first dielectric layer and a first hydrophobic layer, the drive electrode (400) being for controlling the movement of the droplet (500);

and/or;

a second protection layer is arranged on one side of the top plate (201) close to the substrate (101); the second protective layer comprises at least a conductive layer and a second hydrophobic layer located on a side of the conductive layer near the substrate (101).

9. The microfluidic chip according to claim 2, further comprising a temperature control member (103) connected to the substrate (101), the temperature control member (103) being configured to adjust the temperature of the substrate (101) to control the temperature within the sealed cavity to 4-97 ℃.

10. A detection device comprising the microfluidic chip of any one of claims 1-9, further comprising a light source (600); and

an output module (700) comprising a first output connected to the drive electrode (400) and a second output connected to the sensing component (200); the output module (700) is capable of outputting different voltage drive signals to the electrical connection path;

the image processing module (800) is used for collecting the morphology of the liquid drop (500) to be detected in the channel under different voltage driving signals and detecting the contact angle of the liquid drop (500) to be detected.

11. The detection device according to claim 10, wherein the driving electrodes (400) are patterned and arranged on the substrate (101) at intervals, and the control unit further controls the output module (700) to output a corresponding voltage signal to the driving electrodes (400) to control the movement of the droplet (500) to be detected.

12. The detection device according to claim 11, characterized in that the image processing module (800) comprises an imaging device (801) for acquiring the morphology of the drop (500) to be detected; and a main control device (802) connected to the image pickup device (801) and configured to analyze and process the image data.

13. A detection method using the detection device according to any one of claims 10-12, characterized in that the sensing means (200) comprises a top plate (201) movably arranged with the base plate (101) and at least one sensing electrode (202) arranged on the top plate (201), the top plate (201) and the base plate (101) being spaced apart to form the channel; the detection method at least comprises the following steps:

a. providing a drop (500) to be measured into the channel;

b. after the top plate (201) is driven to move to an initial position relative to the base plate (101) by using a control device, detecting the position information of the sensing electrode (202) by using a positioning device and feeding back the position information to the control part, wherein the control part further adjusts the position of the sensing electrode (202) based on the position information, so that the end part of the sensing electrode (202) is contacted with a liquid drop (500) to be detected, and the sensing electrode (202) and the driving electrode (400) form an electric connection path through the liquid drop (500) to be detected;

c. and applying voltages to the driving electrode (400) and the sensing component (200) by using an output module (700), and acquiring a drop angle and a morphology change chart of the drop (500) to be detected under corresponding conditions by using an image processing module (800).

14. The detection method of a detection device according to claim 13, wherein in step a at least two of said droplets (500) to be detected having different sizes are located in said channel, step b further comprises: and respectively adjusting the distances between the sensing electrodes (202) corresponding to different liquid drops (500) to be detected and the corresponding liquid drops (500) to be detected.

15. A sequencing device comprising a microfluidic chip according to any one of claims 1 to 9.

16. A method for manufacturing a droplet control chip for a sequencer, comprising determining process parameters of the droplet control chip by using the detection device according to claim 13, wherein the process parameters at least comprise position information and voltage control parameters; the sensing component (200) comprises a base plate (101), a top plate (201) which is movably arranged and at least one sensing electrode (202) arranged on the top plate (201), wherein the channel is formed between the top plate (201) and the base plate (101) at intervals; determining the process parameters of the liquid drop control chip comprises:

selecting a droplet (500) to be measured with a specified size, and applying the droplet (500) to be measured into the channel;

Adjusting the relative distance between the top plate (201) and the substrate (101), and further adjusting the position between the sensing electrode (202) and the droplet (500) to be detected, and acquiring the position information corresponding to the top plate, the substrate (101), the sensing electrode (202) and the driving electrode (400) when the end part of the sensing electrode (202) is in contact with the droplet (500) to be detected;

adjusting the output module (700) to output different voltage driving signals to the electric connection path, and acquiring voltage control information corresponding to the liquid drop (500) to be detected;

and determining the optimal voltage control parameters based on the morphology and the contact angle of the liquid drop (500) to be detected under different voltage driving signals detected by the image processing module.