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CN121347804A - System for forming a microarray of analyte determination areas on a solid support - Google Patents

System for forming a microarray of analyte determination areas on a solid support

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Publication number
CN121347804A
CN121347804A CN202511772809.0A CN202511772809A CN121347804A CN 121347804 A CN121347804 A CN 121347804A CN 202511772809 A CN202511772809 A CN 202511772809A CN 121347804 A CN121347804 A CN 121347804A
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China
Prior art keywords
liquid
gas
pressure
valve
liquid suction
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CN202511772809.0A
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Chinese (zh)
Inventor
仝南翔
李官宁
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Shandong Haina Data Technology Co ltd
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Shandong Haina Data Technology Co ltd
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Abstract

A system for forming a microarray of analyte measurement areas on a solid support includes a liquid suction gas path, a liquid spray gas path, and a valve assembly configured to allow gas to flow from a liquid suction head to a liquid suction tube in a liquid suction state and to allow gas to form a pulsed high pressure gas in the liquid suction tube in a liquid spray state. According to the technical scheme provided by the invention, the detection object solution can be printed into the reagent tray hole in the form of nano-scale and other liquid drops, and micro detection points with the diameter of micron level are formed at the bottom of the tray body, so that the use amount of detection antibodies/antigens is reduced in geometric magnitude, and the cost is reduced by tens of times of that of other detection products. The miniature monitoring points printed by the technical scheme of the invention have clear boundaries, are nearly circular, have uniform content distribution and have no halation staining blocks around.

Description

System for forming a microarray of analyte determination areas on a solid support
Technical Field
The present invention is in the field of simultaneous analysis of a large number of very small biological samples, and in particular relates to a system for printing on a substrate a micro-dot array for use in the biotechnology industry.
Background
Multiplex immunoassay technology is currently one leading edge method for simultaneous detection and analysis of multiple substances. The method is based on the biochemical reaction principle of the traditional enzyme-linked immunosorbent assay (ELISA) method, can detect multiple biomarkers of a certain disease or specific biomarkers of a plurality of diseases at the same time with high flux, and overcomes the clinical sensitivity and specificity limitation of a single marker. Compared with a plurality of single analysis methods, the multiplex immunoassay technology reduces reagent consumption, reduces manual operation, accelerates analysis speed, and greatly improves the efficiency of detecting and analyzing various cytokines.
The solid-phase planar array method is to fix molecules or microsphere carriers capable of binding to target substances on a solid-phase plane by deposition, external force or a modification method, so that each array point on the plane can be used as a probe of a specific target molecule. Such as immobilization of antibody protein modifications on a solid phase plane to form planar antibody arrays, parallel analysis of multiple targets in a given sample, these types of antibody arrays have become tools for the exploration and study of protein abundance, function, pathways, and potential drug targets. Solid-phase planar arrays can be designed to carry multiple characteristic planes and thus can produce high-dimensional data, which can exhibit many advantages over conventional, single-detection methods, such as higher throughput, higher sensitivity, lower sample volumes, etc.
There are a number of methods currently used to fabricate arrays of biological macromolecules. Such as by heating the membrane or exposing it to ultraviolet radiation, the detection object is immobilized on the porous membrane, however, this is a manual process which is only suitable for manufacturing one array at a time, with a limited number of samples per array, typically 96 samples, which is insufficient for applications requiring analysis of thousands of samples. A more efficient technique is to use a set of steel needles immersed in the wells of a 96-well microplate, and transfer the samples through the needles onto the substrate, thereby producing an ordered array. Khrapko et al spot the test object manually onto the polyacrylamide film using a micropipette. The limitation of the above approach is that the volume of each pixel in the array is large and the difference is significant. The ink jet printing and other "drop on demand" devices for the manufacture of biochemical arrays as disclosed in chinese patent CN118067987a require large amounts of sample and reagents for analysis, it is difficult to perform mass screening of samples and the quality of the resulting microarray cannot be controlled, and in addition, there are disadvantages of inaccuracy and poor durability in the sample sucking and transferring process.
All of the above methods and apparatus are not suitable for mass production of microarrays having (i) a large number of tiny analysis areas with final detection spots of micron-scale or less in diameter, (ii) a specific amount of analyte in each array area, the detection solution being printed in nanograded droplets into the wells of the reagent disk to geometrically reduce the amount of detection antibody/antigen used, and (iii) no pathological imaging of the spots, such as satellite or orthodontic.
However, the performance of multiplex immunoassays is closely related to the above-described features of microarrays, particularly in terms of dot imaging results and the amount of analyte used, and there is an increasing demand in the medical field for an analyte detection system that is cost-effective and accurate. Thus, it would be particularly advantageous to provide a system for forming a microarray of analyte determination areas on a solid support that enhances the reliability and quantification of the results of the analysis, in terms of rapid identification of disease specificity.
Disclosure of Invention
The present invention aims to solve, at least to some extent, one of the technical problems existing in the prior art, and to this end, it provides a system for forming a microarray of analyte determination areas on a solid support.
According to one aspect of the present invention, there is provided a system for forming a microarray of an analyte measurement area on a solid support, comprising a pipetting channel on which a pipetting device is provided, the pipetting device comprising a pipetting head and a pipette, an outlet of the pipetting head being in communication with an inlet of the pipette, the pipetting channel being configured to temporarily store a liquid sample collected by the pipetting head in the pipette, a spraying channel, both ends of the spraying channel being in communication with a high pressure gas outlet and the pipette, respectively, the spraying channel being configured to introduce the high pressure gas into the pipette and to spray the sample temporarily stored in the pipette onto the solid support, and a valve assembly configured to allow gas to flow from the pipetting head to the pipette in a pipetting state and to allow gas to form a pulsed high pressure gas in the pipette in a spraying state.
Preferably, the valve assembly comprises a first solenoid valve having an air inlet in communication with the high pressure gas outlet, an air outlet in communication with a first port of a first three-way valve having an air outlet in communication with the wicking means, a second port of the first three-way valve in communication with a first one-way valve, and a third port of the first three-way valve in communication with the negative pressure pump, wherein the first one-way valve is configured to allow only gas to be directed out of the spray gas path.
Preferably, the first electromagnetic valve is communicated with the liquid suction device through a second electromagnetic valve, the air outlet of the first electromagnetic valve is communicated with the air inlet of the second electromagnetic valve, the air outlet of the second electromagnetic valve is communicated with the atmosphere through an air leakage pipe, and the air outlet of the second electromagnetic valve is communicated with the liquid suction device.
Preferably, the second electromagnetic valve is communicated with the liquid suction device through a second three-way valve, an air outlet of the second electromagnetic valve is communicated with a first port of the second three-way valve, a second port of the second three-way valve is communicated with a second one-way valve, and a third port of the second three-way valve is communicated with the liquid suction device, wherein the second one-way valve is configured to only allow gas to be led out from the liquid spraying gas path.
Preferably, the high-pressure gas is provided through a gas source subsystem and a pressure stabilizing subsystem, the gas source subsystem comprises a high-pressure gas source for creating a high-pressure environment, the pressure stabilizing subsystem comprises a pressure stabilizing cabin for creating a stable and controllable working target pressure, and a gas inlet of the first electromagnetic valve is communicated with a high-pressure gas outlet of the pressure stabilizing cabin.
Preferably, the liquid injection device further comprises a capacity expansion cavity which is connected in series on the liquid injection gas path and is positioned between the second electromagnetic valve and the second three-way valve.
Preferably, a first flow control valve is arranged at the outlet of the high-pressure air source and used for controlling the air flow in the liquid spraying air path.
Preferably, a second flow control valve is arranged at the outlet of the negative pressure pump and is used for controlling the gas flow of the liquid suction gas channel.
Preferably, the gas source subsystem further comprises a gas filter.
Preferably, the liquid suction head and the liquid suction pipe are used for holding the sucked sample, so that the liquid level of the sample can be ensured to be in stable contact with the high-pressure air flow pulse, and meanwhile, the form of liquid drops is ensured to meet the requirements of the micro-point array and have repeatability through the micron-sized opening of the tip.
Compared with the prior art, the invention has the advantages that the technical scheme can print the detection object solution into the holes of the reagent tray in the form of nano-scale modified liquid drops, and micro detection points with the diameter of micron level are formed at the bottom of the tray body, so that the use amount of detection antibodies/antigens is reduced in geometric magnitude, and the cost is reduced by tens of times of that of other detection products. The miniature monitoring points printed by the technical scheme of the invention have clear boundaries, are nearly circular, have uniform content distribution and have no halation staining blocks around.
Drawings
The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this specification, illustrate embodiments of the application and together with the description serve to explain the application and do not constitute a limitation on the application. In the drawings:
FIG. 1 illustrates a system for forming a microarray of analyte determination areas on a solid support according to an embodiment of the present invention;
FIG. 2 illustrates a Kong Nadian morphology of the components provided in accordance with an embodiment of the present invention when performing operations using the system shown in FIG. 1;
FIG. 3 illustrates a system for forming a microarray of analyte determination areas on a solid support according to an embodiment of the present invention;
FIG. 4 illustrates a Kong Nadian morphology of the components provided in accordance with an embodiment of the present invention when performing operations using the system shown in FIG. 3;
FIG. 5 illustrates a system for forming a microarray of analyte determination areas on a solid support according to an embodiment of the present invention;
FIG. 6 is a diagram showing a Kong Nadian morphology of components of an operation performed using the system shown in FIG. 5, according to an example embodiment of the present invention;
FIG. 7 illustrates a system for forming a microarray of analyte determination areas on a solid support according to an embodiment of the present invention;
FIG. 8 illustrates a Kong Nadian morphology of the components provided in accordance with an embodiment of the present invention when performing operations using the system illustrated in FIG. 7.
Fig. 9 shows a structure of a pipette and a pipette head provided according to an embodiment of the present invention.
Detailed Description
The examples set forth below are presented to provide those skilled in the art with a more clear understanding of the present invention. The following examples are not intended to limit the scope of the invention, but are merely illustrative examples. The starting materials, reagents or apparatus mentioned in the examples below are all available commercially, or are obtained by known existing means, unless otherwise specified.
In this specification, the terms "first," "second," and the like are used not to order or denote importance or a primary or secondary relationship, but rather to distinguish one element from another.
According to the general inventive concept of the present disclosure, there is provided a system for forming an analyte measurement area microarray on a solid support, including a pipetting channel on which a pipetting device is provided, the pipetting device including a pipetting head and a pipette, an outlet of the pipetting head being in communication with an inlet of the pipette, the pipetting channel being configured to temporarily store a liquid sample collected by the pipetting head in the pipette, a spraying channel having both ends respectively in communication with a high-pressure gas outlet and the pipette, the spraying channel being configured to introduce the high-pressure gas into the pipette and spray the sample temporarily stored in the pipette onto the solid support, and a valve assembly configured to allow gas to flow from the pipetting head to the pipette in a pipetting state and to allow gas to form a pulsed high-pressure gas in the pipette in a spraying state. The length of the liquid suction pipe is 13.23mm, the outer diameter is 2.60mm, the inner diameter is 1.60 mu m, the length of the liquid suction head is 1.73mm, the diameter of a hole is 40 mu m, the convergence angle of the inner side wall of the liquid suction head is 54 degrees, and the dead liquid amount can be ensured to be smaller than 20 nanoliters through verification. The inner side of the hole is treated by a silicon-based super-hydrophobic nano coating, so that the surface tension of a printing solution and the pipe wall is reduced, and the adhesion of residual liquid is reduced. The whole liquid suction pipe and the liquid suction head adopt integral 3D printing so as to avoid cross contamination caused by residual liquid remaining at the meshing part of the components.
Example 1
FIG. 1 is a system for forming a microarray of analyte determination areas on a solid support according to one embodiment of the present disclosure. In this embodiment, the system comprises a gas source subsystem, a voltage stabilizing subsystem and a solenoid valve 101, wherein the gas source subsystem comprises a high-pressure gas source and a gas filter and is used for creating a high-pressure environment, the gas source pressure is lifted until the gas source pressure is higher than a target working pressure, the target working pressure varies from 50KPa to 200KPa according to the executed task, the voltage stabilizing subsystem comprises a voltage stabilizing and electric control high-precision voltage stabilizing valve and a voltage stabilizing cabin and is used for creating a stable and controllable working target pressure, a gas inlet A of the solenoid valve 101 is communicated with a high-pressure gas outlet (a voltage stabilizing cabin gas outlet), a gas outlet C of the solenoid valve 101 is communicated with a liquid suction device through an expansion cavity, the liquid suction device comprises a liquid suction head and a liquid suction pipe, the outlet of the liquid suction head is communicated with the inlet of the liquid suction pipe, and a gas outlet B of the solenoid valve 101 is communicated with a negative pressure liquid suction pump. After the electromagnetic valve 101A is closed, the passages 101B-C are opened, and the liquid sample collected by the liquid suction head is temporarily stored in the liquid suction pipe under the action of the negative pressure liquid suction pump. After the electromagnetic valve 101A is opened, the 101A-C passage is opened, high-pressure gas at the left end of the A side is flushed into a pipeline and an expansion cavity in front of the spray head through the C end in the opening time, the high-pressure gas is guided into the liquid suction pipe by the liquid spraying gas passage mechanism, and the sample temporarily stored in the liquid suction pipe is sprayed onto the solid carrier.
The sample composition comprises three types, namely a component one with lower viscosity of 0.9 mPa.s (temperature of 25 ℃ C., water+0.1% v/v Tween 20+pigment), a component two with higher viscosity of 1.3 mPa.s (temperature of 25 ℃ C., 15% v/v glycerol aqueous solution+pigment) and a component three with higher viscosity of 2.0 mPa.s (temperature of 25 ℃ C., 30% v/v glycerol aqueous solution+pigment). The liquid volume of the single spray in the pipette and the pipette head can be stabilized to a certain set value (CV < 7%) in the range of 2-50 nanoliters, as an example, the pipette and the pipette head can take the structure shown in FIG. 9 but are not limited to the example, (the tip opening of the pipette head is 40 micrometers, and the pipette head is printed by using polyethylene glycol diacrylate material through 3D light curing, and has high biocompatibility, and the port printing resolution is 2 micrometers, as an example, the solid carrier adopts transparent polystyrene material subjected to surface treatment, the surface treatment mode comprises plasma spraying, chemical solution soaking and the like, as an example, the target working pressure can be varied from 50KPa to 200KPa, the target working pressure is 100KPa, as shown in FIG. 2, and FIG. 2 shows the shape of Kong Nadian when the needle head is operated under the second gas path component, and pigment is used for coloring, as a result of the figure, the satellite points in each hole are more due to accumulated liquid, and the accumulated pressure in the gas path is gradually accumulated, the point shape is deformed under the design, the accumulated pressure is released, and the needle head is rapidly accumulated, and the needle head is almost completely deformed when the needle head is operated under the design.
Example two
Fig. 3 is a system for forming a microarray of analyte determination areas on a solid support according to one embodiment of the present disclosure. The system comprises a gas source subsystem, a pressure stabilizing subsystem and a solenoid valve 101, wherein the gas source subsystem comprises a high-pressure gas source and a gas filter and is used for creating a high-pressure environment and lifting gas source pressure until the gas source pressure is higher than target working pressure, the pressure stabilizing subsystem comprises a stable electric control high-precision pressure stabilizing valve and a pressure stabilizing cabin and is used for establishing stable and controllable working target pressure, a gas inlet A of the solenoid valve 101 is communicated with a high-pressure gas outlet (a pressure stabilizing cabin gas outlet), a gas outlet C of the solenoid valve 101 is communicated with a liquid suction device through a capacity expansion cavity, the liquid suction device comprises a liquid suction head and a liquid suction pipe, the outlet of the liquid suction head is communicated with the inlet of the liquid suction pipe, a gas outlet B of the solenoid valve 101 is communicated with a first end of a tee joint 201, a second end of the tee joint 201 is communicated with a one-way air release valve 1, and a third end of the tee joint 201 is communicated with a negative pressure liquid suction pump. After the electromagnetic valve 101A is closed, the passages 101B-C are opened, and the liquid sample collected by the liquid suction head is temporarily stored in the liquid suction pipe under the action of the negative pressure liquid suction pump. After the electromagnetic valve 101A is opened, the 101A-C passage is opened, high-pressure gas at the left end of the A side is flushed into a pipeline and an expansion cavity in front of the spray head through the C end in the opening time, the high-pressure gas is guided into the liquid suction pipe by the liquid spraying gas passage mechanism, and the sample temporarily stored in the liquid suction pipe is sprayed onto the solid carrier. After the electromagnetic valve 101A is closed, the oscillation is generated in the pipeline under the combined action of the one-way air release valve 1 and the temporary interface of the liquid suction head.
Printing was performed with a sample having a viscosity of 0.9 mPas (temperature 25 ℃ C., water+0.1% v/v Tween20+ pigment). The liquid drop volume in the pipette and pipette head was 10 nanoliters. The target operating pressure is 100Kpa. The ejection results are shown in fig. 4A. Printing was performed with a sample having a viscosity of 1.3 (temperature 25 ℃,15% v/v glycerol in water + pigment). The liquid drop volume in the pipette and pipette head was 10 nanoliters. The target operating pressure is 100Kpa and the injection result is shown in fig. 4B. The ejected microdroplet may experience a succession of distortions.
In the embodiment, a one-way air release valve 1 is added in the pipeline. Under the design of fixed hydrojet head, the shower nozzle can't change fast, and the liquid in the shower nozzle can only be changed through "positive pressure extrusion-negative pressure suction" mode. The exhaust port B of the electromagnetic valve 101 can meet the liquid feeding/changing requirement of the spray head by being connected with a negative pressure liquid suction pump. As shown in the air path of the present embodiment, when the air inlet a end of the solenoid valve 101 is closed, the BC end of the solenoid valve 101 is communicated. At this time, the high-pressure gas in the pipeline is discharged to the outside atmosphere through the unidirectional air release valve when impacting the liquid level, so that the air pressure in the pipeline is quickly reduced. The gas circuit of the embodiment effectively relieves the phenomenon of effusion of the spray head caused by the accumulation of high-pressure gas in the pipe in the first embodiment. Meanwhile, due to the existence of the one-way air release valve, the air pressure and the pulse duration of the pulse gas are obviously increased, the hardware precision required by maintaining the continuous operation of the system is reduced, and the robustness of the system is improved. However, it was also observed in this example that the ejected microdroplet would exhibit a number of distortions in succession.
Example III
Fig. 5 is a system for forming a microarray of analyte determination areas on a solid support according to one embodiment of the present disclosure. The system comprises a gas source subsystem, a pressure stabilizing subsystem and a solenoid valve 101, wherein the gas source subsystem comprises a high-pressure gas source and a gas filter and is used for creating a high-pressure environment and lifting gas source pressure until the gas source pressure is higher than target working pressure, the pressure stabilizing subsystem comprises a stable electric control high-precision pressure stabilizing valve and a pressure stabilizing cabin and is used for establishing stable and controllable working target pressure, a gas inlet A of the solenoid valve 101 is communicated with a high-pressure gas outlet (a pressure stabilizing cabin gas outlet), a gas outlet C of the solenoid valve 101 is communicated with a liquid suction device through a capacity expansion cavity, the liquid suction device comprises a liquid suction head and a liquid suction pipe, the outlet of the liquid suction head is communicated with the inlet of the liquid suction pipe, a gas outlet B of the solenoid valve 101 is communicated with a first end of a tee joint 201, a second end of the tee joint 201 is communicated with a one-way air release valve 1, and a third end of the tee joint 201 is communicated with a negative pressure liquid suction pump. After the electromagnetic valve 101A is closed, the passages 101B-C are opened, and the liquid sample collected by the liquid suction head is temporarily stored in the liquid suction pipe under the action of the negative pressure liquid suction pump. After the electromagnetic valve 101A is opened, the opening time is in the ms level, the 101A-C passage is opened, high-pressure gas at the left end of the 101A side is flushed into the pipeline and the expansion cavity before the nozzle through the 101C end in the opening time, the liquid spraying gas passage mechanism guides the high-pressure gas into the liquid suction pipe, the sample temporarily stored in the liquid suction pipe is sprayed onto the solid carrier, and after the electromagnetic valve 101A is closed, the one-way air release valve 1, the temporary air interface of the liquid suction head and the one-way air release valve 2 jointly act to enable oscillation to be generated in the pipeline.
Printing was performed with a sample having a viscosity of 1.3 (temperature 25 ℃,15% v/v glycerol in water + pigment). The liquid drop volume in the pipette and pipette head was 10 nanoliters. The target operating pressure is 100Kpa. The ejection results are shown in fig. 6, in which the ejected microdroplets have random single-dot morphological instability.
Example IV
Fig. 7 is a system for forming a microarray of analyte determination areas on a solid support according to one embodiment of the present disclosure. The system in this embodiment includes a gas supply subsystem including a high pressure gas supply and a gas filter for creating a high pressure environment, raising the gas supply pressure above a target operating pressure (the gas supply is a gas pump, the maximum output pressure is about 0.8mpa. The pressure entering the system is 100Kpa in this embodiment regulated by a pressure regulating subsystem); the system also comprises a voltage stabilizing subsystem, wherein the voltage stabilizing subsystem comprises a voltage stabilizing control high-precision voltage stabilizing valve and a voltage stabilizing cabin, and is used for establishing stable and controllable working target pressure, an air inlet A of an electromagnetic valve 101 (the model is Feston brand MHE3-MS1H-3/2G-QS-6-K, parameters are shown in the following table) is communicated with a high-pressure air outlet (a voltage stabilizing cabin air outlet), an air outlet C of the electromagnetic valve 101 is communicated with a liquid suction device through the electromagnetic valve 105, an expansion cavity and a tee joint 202 (the expansion cavity can enlarge the amplitude of Helmholtz resonance, and meanwhile, the air pressure of the high-pressure air can be correspondingly reduced along with the volume increase in the expansion cavity, so that the higher input pressure can be used, for example, a certain liquid can be stably printed by adopting a parameter combination of 20kpa+1MS, but uniform air pulse under the parameter cannot be accurately realized due to the fact that the pipeline precision is adopted, for example, a volume cabin can be used, for example, 50 ml of the required parameter can be printed by a large-volume expansion cavity can not be printed by a so-called a high-precision printing system, and a large-volume expansion cavity can be printed by a large-volume printing system is a well-free container, and a large-free expansion cavity is formed by a container is a container, and a large-free expansion system is required to be printed by a large-quality amplification system, and is a container is made of a material is a container, because is not has a stable container, because is required to be printed, all can finish tasks), the liquid suction device comprises a liquid suction head and a liquid suction pipe, an outlet of the liquid suction head is communicated with an inlet of the liquid suction pipe, an exhaust port B of the electromagnetic valve 101 is communicated with a first end of the tee joint 201, a second end of the tee joint 201 is communicated with the one-way air release valve 1, and a third end of the tee joint 201 is communicated with the negative pressure liquid suction pump. After solenoid valve 101A and solenoid valve 102A are closed, 101B-C and 102B-C are opened, and the liquid sample collected by the pipette head is temporarily stored in the pipette under the action of the negative pressure pipette. After the electromagnetic valve 101A and the electromagnetic valve 102A are opened, the opening time is in the ms level, the 101A-C passage and the 102A-C passage are opened, high-pressure gas at the left end of the 101A side is flushed into a pipeline and an expansion cavity before the nozzle through the 101C end and the 102A-C passage in the opening time, the high-pressure gas is introduced into the liquid suction pipe by the liquid spraying gas passage mechanism, a sample temporarily stored in the liquid suction pipe is sprayed onto a solid carrier, and after the electromagnetic valve 101A and the electromagnetic valve 102A are closed, the combined action of the one-way air release valve 1, the temporary air interface of the liquid suction head, the air release pipe and the one-way air release valve 2 enables the sectional control air pressure pulse to reduce the accumulated liquid.
TABLE 1 MHE3-MS1H-3/2G-QS-6-K parameters
The sample composition comprises three types, namely a component one with lower viscosity of 0.9 mPa.s (temperature of 25 ℃ C., water+0.1% v/v Tween 20+pigment), a component two with higher viscosity of 1.3 mPa.s (temperature of 25 ℃ C., 15% v/v glycerol aqueous solution+pigment) and a component three with higher viscosity of 2.0 mPa.s (temperature of 25 ℃ C., 30% v/v glycerol aqueous solution+pigment). The volume of liquid in a single shot in the pipette and pipette head is 2-50 nanoliters. The target operating pressure varies from 50KPa to 200KPa depending on the task being performed. The results of the ejection using component 2 as an example are shown in fig. 8 (the morphology of the ejection results of component 1 and component 3 is identical to that of component 2, omitted here), and pigment coloring is used. The picture shows that the points in each hole have clear boundaries, are nearly circular, have uniform content distribution and have no halation staining blocks around. Experiments prove that the design can effectively complete the micro-droplet printing task, and the selectable droplet capacity is 2-15 nanoliters. The printed liquid drops are full and round in shape on the working surface, extremely high in uniformity and repeatability and strong in robustness, and can be used for mass industrial production.
The following data is a statistical plot of the dot morphology state under each pipeline system, at 22 degrees ambient, using 25% v/v glycerol aqueous solution and pigment, due to the influence of the gas path injection system. There may be a manual count error. In each gas path system, 1000 dots are printed consecutively. Specifically, a 25 dot per well (5×5) matrix was printed with 40 wells. The printing pulse pressure and pulse duration have been optimized separately for each system. Printing is carried out on a standard transparent PS 96-well ELISA plate. The experimental results of the air circuit of the above embodiment are summarized as follows.
The times of needle head effusion which is necessary to be manually intervened are that obvious needle head effusion is observed by naked eyes, the point form on the working surface is obviously deformed or obvious satellite points exist, and continuous spraying of liquid is not recovered for more than 10 times. The number of times that the automatically recoverable needle head dropsy is observed is that the obvious needle head dropsy is observed by naked eyes, the shape on the working surface is obviously deformed or obvious satellite points exist, but the normal working state is recovered within 10 liquid spraying times (sucking back or large liquid drop spraying).
Satellite splash refers to a splash point with a distinct unexpected printed dot around the target point. Too large and too small means that the size of the target point observed by naked eyes deviates significantly from the expectation. By significantly non-circular is meant that the morphology of a point is significantly non-circular, possibly elliptical, spindle, semi-circular, or other shaped. The offset position means that the position of a certain point is significantly offset from the expected matrix print position and it can be judged that it is not caused by the moving platform (non-systematic offset).
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1.一种在固体载体上形成分析物测定区域微阵列的系统,其特征在于,包括:1. A system for forming a microarray of analyte measurement regions on a solid support, characterized in that it comprises: 吸液气路,所述吸液气路上设置有吸液装置,所述吸液装置包括吸液头和吸液管,所述吸液头的出口与所述吸液管的入口连通,所述吸液气路构造成将所述吸液头所采集的液体样品暂存在所述吸液管内;A liquid aspiration gas path is provided, wherein a liquid aspiration device is provided on the liquid aspiration gas path, the liquid aspiration device includes a liquid aspiration head and a liquid aspiration tube, the outlet of the liquid aspiration head is connected to the inlet of the liquid aspiration tube, and the liquid aspiration gas path is configured to temporarily store the liquid sample collected by the liquid aspiration head in the liquid aspiration tube; 喷液气路,所述喷液气路的两端分别与高压气体出口及所述吸液管连通,所述喷液气路构造成将所述高压气体导入所述吸液管内,并将暂存在所述吸液管内的样品喷射至所述固体载体上;The liquid spraying gas path has two ends connected to a high-pressure gas outlet and the liquid suction tube, respectively. The liquid spraying gas path is configured to introduce the high-pressure gas into the liquid suction tube and spray the sample temporarily stored in the liquid suction tube onto the solid carrier. 阀组件,所述阀组件构造成在吸液状态下允许气体从所述吸液头流向所述吸液管,在喷液状态下允许气体在所述吸液管中形成脉冲高压气体。A valve assembly configured to allow gas to flow from the suction head to the suction tube in a suction state, and to allow gas to form a pulsed high-pressure gas in the suction tube in a spray state. 2.根据权利要求1所述的系统,其特征在于,所述阀组件包括第一电磁阀,所述第一电磁阀的进气口与高压气体出口连通,所述第一电磁阀的排气口与第一三通阀的第一端口连通,所述第一电磁阀的出气口与所述吸液装置连通,所述第一三通阀的第二端口与第一单向阀连通,所述第一三通阀的第三端口与负压泵连通,其中,所述第一单向阀构造成仅允许从所述喷液气路中导出气体。2. The system according to claim 1, wherein the valve assembly includes a first solenoid valve, the inlet of the first solenoid valve being connected to a high-pressure gas outlet, the outlet of the first solenoid valve being connected to a first port of a first three-way valve, the outlet of the first solenoid valve being connected to the liquid suction device, the second port of the first three-way valve being connected to a first check valve, and the third port of the first three-way valve being connected to a negative pressure pump, wherein the first check valve is configured to allow gas to be discharged only from the liquid injection gas path. 3.根据权利要求2所述的系统,其特征在于,所述第一电磁阀通过第二电磁阀与所述吸液装置连通,所述第一电磁阀的出气口与所述第二电磁阀的进气口连通,所述第二电磁阀的排气口通过泄气管与大气连通,所述第二电磁阀的出气口与所述吸液装置连通。3. The system according to claim 2, wherein the first solenoid valve is connected to the liquid suction device through the second solenoid valve, the air outlet of the first solenoid valve is connected to the air inlet of the second solenoid valve, the exhaust port of the second solenoid valve is connected to the atmosphere through a vent pipe, and the air outlet of the second solenoid valve is connected to the liquid suction device. 4.根据权利要求2所述的系统,其特征在于,所述第二电磁阀通过第二三通阀与所述吸液装置连通,所述第二电磁阀的出气口与所述第二三通阀的第一端口连通,所述第二三通阀的第二端口与第二单向阀连通,所述第二三通阀的第三端口与所述吸液装置连通,其中,所述第二单向阀构造成仅允许从所述喷液气路中导出气体。4. The system according to claim 2, wherein the second solenoid valve is connected to the liquid suction device via a second three-way valve, the outlet of the second solenoid valve is connected to the first port of the second three-way valve, the second port of the second three-way valve is connected to a second one-way valve, and the third port of the second three-way valve is connected to the liquid suction device, wherein the second one-way valve is configured to allow gas to be discharged only from the liquid spraying gas path. 5.根据权利要求1所述的系统,其特征在于,通过气源子系统和稳压子系统提供所述高压气体;5. The system according to claim 1, characterized in that the high-pressure gas is provided through a gas source subsystem and a pressure stabilizing subsystem; 所述气源子系统包括高压气源,用于创建高压环境;The gas source subsystem includes a high-pressure gas source for creating a high-pressure environment; 所述稳压子系统包括稳压舱,用于建立稳定可控的工作目标压力;The pressure stabilization subsystem includes a pressure stabilization chamber for establishing a stable and controllable target working pressure; 所述第一电磁阀的进气口与所述稳压舱的高压气体出口连通。The air inlet of the first solenoid valve is connected to the high-pressure gas outlet of the pressure stabilizing chamber. 6.根据权利要求1所述的系统,其特征在于,还包括串联在所述喷液气路上且位于所述第二电磁阀和所述第二三通阀之间的扩容腔。6. The system according to claim 1, characterized in that it further includes an expansion cavity connected in series in the liquid injection gas path and located between the second solenoid valve and the second three-way valve. 7.根据权利要求1所述的系统,其特征在于,所述高压气源出口处设置有第一流量控制阀,用于控制所述喷液气路中的气体流量。7. The system according to claim 1, wherein a first flow control valve is provided at the outlet of the high-pressure gas source for controlling the gas flow rate in the liquid injection gas path. 8.根据权利要求1所述的系统,其特征在于,所述负压泵出口处设置有第二流量控制阀,用于控制所述吸液气路的气体流量。8. The system according to claim 1, wherein a second flow control valve is provided at the outlet of the negative pressure pump for controlling the gas flow rate of the liquid suction gas path. 9.根据权利要求1所述的系统,其特征在于,所述气源子系统还包括气体过滤器。9. The system according to claim 1, wherein the gas source subsystem further comprises a gas filter. 10.根据权利要求1所述的系统,其特征在于,所述吸液头和吸液管为一体式3D打印。10. The system according to claim 1, wherein the suction head and the suction tube are integrally 3D printed.
CN202511772809.0A 2025-09-01 2025-11-28 System for forming a microarray of analyte determination areas on a solid support Pending CN121347804A (en)

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