CN220568810U - Paper chip device - Google Patents

Abstract

The utility model relates to the field of real-time detection technology, specifically to a paper chip device. The paper chip device includes a connecting tube and a negative pressure generator. A paper chip is loaded in the cavity of the connecting tube. One end of the connecting tube There is a hollow microneedle connected to it, and the other end is connected to the negative pressure generator during detection. The hollow microneedle is provided with a hole connected to the connecting tube. Under the action of the negative pressure generator, the hollow microneedle can be directly connected from The tissue fluid is extracted subcutaneously. The tissue fluid flows from the hollow microneedle into the connecting tube. The tissue fluid fully contacts the paper chip in the connecting tube. The paper chip is then used to quickly analyze and detect the tissue fluid immediately. The test results can be obtained within 10 minutes.

Description

A paper chip device

Technical field

The utility model relates to the field of real-time detection technology, and specifically relates to a paper chip device.

Background technique

Point-of-care testing devices (POCT devices) can meet the needs of on-site testing, easy operation, rapid monitoring of health status, and even rapid diagnosis of diseases. They have attracted widespread attention and have a huge and rapidly growing market. Global Market. Blood is currently the most common biological fluid sample for POCT, but has many disadvantages associated with invasive sampling. For example, currently commonly used venous blood collection methods or fingertip blood collection methods will cause pain, wounds, and infection risks to patients. Additionally, blood sampling often requires a trained medical professional. Interstitial fluid contains most of the components present in blood, including inorganic salts, physiological small molecules, RNAs, circulating proteins, antibodies, hormones, exosomes, antibiotics, and some diet-related metabolites (such as alcohol, caffeine), etc., all of which are Can be used as a detection target. Moreover, the collection of tissue fluid can be relatively simple, easy to operate, painless and risk-free, making it a good biological sample for a new generation of point-of-care detection devices.

Microneedle has a hollow or solid needle-like structure, and its needle tip size is as small as tens of microns. Originally used for transdermal delivery of drugs and vaccines, it has recently been developed for the painless extraction of subcutaneous tissue fluid in a minimally invasive manner. To date, various materials including silicon, metals and polymers have been used to create microneedles. The tissue fluid extracted by microneedles can be used for health monitoring, detection and disease diagnosis. However, existing detection methods based on microneedle extraction of tissue fluid are time-consuming, inconvenient, and have many operating steps, which often make it difficult to meet the requirements of real-time detection. For example, hydrogel microneedles are currently the most widely used method for tissue fluid extraction. The detection method based on hydrogel microneedle extraction of interstitial fluid relies on the swelling of the hydrogel and the diffusion of interstitial fluid in it to extract interstitial fluid. However, the diffusion process of tissue fluid in hydrogels is slow, often taking tens of minutes, and the extraction volume is low (usually less than 10 microliters), resulting in the inability to achieve rapid detection (it usually takes tens of minutes to display results). In addition, some technical methods place the detection system inside the hydrogel microneedle, which speeds up the detection, but there is a biosafety risk of leakage of chemical reagents.

Therefore, there is an urgent need to provide a detection device or technology that is easy to operate, performs on-site detection, has rapid detection, and has no biosafety risks.

Utility model content

Therefore, the technical problem to be solved by the present invention is to overcome the shortcomings in the prior art of being unable to quickly and easily detect components in tissue fluid on-site to monitor health conditions, thereby providing a paper chip device.

To this end, the utility model provides a paper chip device, which includes a connecting tube and a negative pressure generator. A paper chip is loaded in the lumen of the connecting tube. One end of the connecting tube is connected with a hollow microneedle, and the other end of the connecting tube is connected with a hollow microneedle. During detection, it is connected to the negative pressure generator, and the hollow microneedle is provided with a hole connected to the connecting tube.

The paper chip device provided by the utility model can directly extract tissue fluid from the subcutaneous tissue through the hollow microneedle under the action of the negative pressure generator. The tissue fluid flows from the hollow microneedle into the connecting tube, and the tissue fluid fully contacts the paper chip in the connecting tube. Paper chips are used for rapid analysis and instant detection of tissue fluid, and the test results can be obtained within 10 minutes.

In some preferred embodiments, a cavity table is also provided between the connecting tube and the hollow microneedle. A cavity structure is formed inside the cavity table, and the cavity of the cavity table is in contact with the hollow microneedle. The channels are connected, and the channels of the hollow microneedle are connected with the connecting tube through the cavity table.

The paper chip device provided by the utility model has a continuous channel from the hollow microneedle channel to the cavity of the cavity table, then to the connecting tube cavity, and finally to the negative pressure generator. The cavity structure inside the cavity table is a gathering place for the tissue fluid extracted by the hollow microneedle. It is also a buffer space for transporting the tissue fluid to the outside. This place is connected to the external connecting tube. During use, the hollow microneedle is inserted into the skin. Driven by capillary action and negative pressure, tissue fluid can automatically enter and be stored in the hollow microneedle, cavity table, connecting tube and negative pressure generator. When using this device to extract tissue fluid, press the cavity table to penetrate the hollow microneedle into the skin, and then connect the connecting tube to the negative pressure generator to form a sequence from "hollow microneedle channel - cavity table cavity - connecting tube lumen - After the continuous channel of the "negative pressure generator", there is no need for any manual operation to collect tissue fluid. You only need to wait for the tissue fluid to automatically enter the cavity table, connecting tube and negative pressure generator through the hollow microneedle.

In some preferred embodiments, one side of the cavity table is provided with a connecting part that communicates with the connecting pipe, and the cavity of the cavity table is connected with the connecting pipe through the connecting part. The connection part is arranged to facilitate the connection between the cavity table and the connecting pipe, so that the cavity table and the connecting pipe can be made of different materials. For example, the connecting pipe can be made of plastic elastic material, while the cavity table can be made of hard material, which improves the connection between the two. The sealing property.

In certain preferred embodiments, the hollow microneedles are provided with multiple channels. The holes on the hollow microneedle are set on the surface of the hollow microneedle and penetrate the bottom surface of the cavity platform to communicate with the cavity of the cavity platform. The multiple holes on the hollow microneedle can increase the extraction speed of tissue fluid while avoiding the clogging of a single hole. This may lead to reduced tissue fluid extraction efficiency or even failure.

In the present invention, the cross section of the cavity table is square, oval, or circular, or a combination of square, oval, and circular. The top surface of the cavity table has a flat, convex, or concave structure. The top surface of the cavity table may be a flat, convex, or concave structure when viewed from the side.

In some preferred embodiments, the height of the cavity structure is 0.5-2.5 mm. Preferably it is 1 mm.

In some preferred embodiments, the bottom surface of the cavity table is a planar structure. Although the cavity table has a planar structure, when the cavity table is pressed to cause the microneedles to penetrate into the skin and during the tissue fluid extraction process, the cavity table can still closely fit the skin.

More preferably, the bottom surface of the cavity table has a flexible or deformable structure. The bottom surface is a flexible or deformable structure. When the cavity table is pressed to allow the microneedle to penetrate the skin and during the tissue fluid extraction process, the cavity table can fit the skin more closely.

In some preferred embodiments, the bottom surface of the cavity table is a curved structure, which is flexible or deformable. When the cavity table is pressed to cause the microneedles to penetrate into the skin and during the tissue fluid extraction process, the cavity table with a curved bottom surface can still closely fit the skin.

In the present invention, the number of the connecting parts is several. Several through-connections can be provided on the side of the cavity table for connecting different connecting pipes and negative pressure generators respectively, so that one cavity table device can be equipped with several sets of negative pressure generators. This utility model is based on micro The needle's negative pressure device is capable of quickly extracting and storing tissue fluid.

More preferably, the number of the connecting parts is 1-4.

In some preferred embodiments, the connecting portion is a cylindrical structure. The connecting pipe is nested and connected to the connecting part. The connecting pipe is a hose and is nested outside the connecting part.

In some preferred embodiments, the connecting tube is a through-tube with openings at both ends, and a hollow connecting tube lumen is located between the openings at both ends.

The inner diameter of the connecting pipe is 1-10mm.

In some preferred embodiments, a support body connecting the top and bottom of the cavity table is provided in the cavity.

The function of the support body is to support the cavity structure to prevent the deformation of the cavity platform structure from causing the cavity to close during the production process, storage process and use process, causing the opening of the hollow microneedle below to be blocked, or to converge into the cavity. The tissue fluid in the cavity structure is blocked, which affects the extraction of tissue fluid. At the same time, the support body can also reduce the dead volume of the cavity in the cavity table and reduce the residual tissue fluid extracted through the hollow microneedle in the cavity of the cavity table.

In some preferred embodiments, the support body is arranged to avoid the opening of the hollow microneedle. The main purpose of avoiding the opening is to prevent the support body from blocking the hole of the hollow microneedle and interfering with the extraction of subcutaneous body fluid, so that the tissue fluid can smoothly pass to the connecting tube.

Preferably, the number of the supports is 2-4.

Preferably, the supports are arranged parallel to each other or around the center of the cavity.

More preferably, the supports are not connected to each other. More preferably, the support body and the inner wall of the cavity are not connected to each other.

Preferably, the support body is provided with a through hole. The through holes on the support allow the subcutaneous tissue fluid in the cavity to flow and do not block the tissue fluid extracted through the hollow microneedle.

More preferably, a plurality of hollow microneedles are provided to form a microneedle array.

Preferably, the tip circumference of the hollow microneedle is smaller than the bottom circumference of the hollow microneedle.

Preferably, the shapes of the hollow microneedles include but are not limited to cones, square cones, pyramids, multiple cones, multiple square cones, multiple pyramids, or combinations of the above shapes.

In some preferred embodiments, the hollow microneedles are an array of hollow microneedles composed of 5 × 5 to 20 × 20 hollow microneedles, with an area of 0.28 to 46.24 cm ² . The microneedle array at this scale takes into account the extraction capacity of the hollow microneedles and the size of the patch, and the extraction capacity meets the extraction requirements of the test.

In some preferred embodiments, the length (or height) of the hollow microneedle is 300-1200 μm. The hollow microneedle with this length is not enough to contact the nerve endings in the dermis, causing little trauma to human skin and reducing pain.

In some preferred embodiments, the center distance between the hollow microneedles and adjacent hollow microneedles is 300 to 3000 μm. This center distance can adapt to the device size requirements in different scenarios.

In some preferred embodiments, the number of holes on each hollow microneedle is 1 to 4.

In some preferred embodiments, the inner diameter of the pore channel is 10-1000 μm.

Preferably, it also includes a storage device, and the negative pressure generator and the storage device can be integrated devices or separate devices.

Preferably, the integrated device includes but is not limited to an integrated device that is pre-evacuated to different degrees of vacuum and uses its negative pressure to collect and store samples at the same time. More preferably, the integrated device for generating and storing negative pressure includes, but is not limited to, a vacuum sampling tube, a vacuum blood collection tube, a decompression sampling tube, or a decompression blood collection tube. When using, first press the extraction end to make the hollow microneedle penetrate the skin; then insert the needle end of the connecting tube into the vacuum sampling tube, or vacuum blood collection tube, or decompression sampling tube, or decompression blood collection tube, and use the vacuum sampling tube to provide The negative pressure can quickly extract and transfer subcutaneous body fluid from under the skin to the connecting tube and vacuum sampling tube, or vacuum blood collection tube, or decompression sampling tube, or decompression blood collection tube.

Optionally, the separate device includes, but is not limited to, a device having a reservoir in front of the negative pressure generating device. The negative pressure generation and storage separation device refers to a device that drives the sample flow and extracts the sample through the negative pressure space, and stores the sample on the channel where the negative pressure drives the sample flow. This storage space is separated from the space of the negative pressure generator. rather than the same space.

Preferably, the height of the cavity is 0.5-2.5mm.

The materials of the cavity table and hollow microneedles include but are not limited to organic polymers and inorganic materials.

More preferably, the inorganic materials include metals and inorganic non-metallic materials.

More preferably, the organic polymers include, but are not limited to, methacrylated terminal polymers such as methacrylated gelatin, methacrylated hyaluronic acid, methacrylated polyethylene glycol, etc. , polydimethylsiloxane, polyvinyl alcohol, polyurethane, polyethylene glycol, light-curing resin.

More preferably, the photocurable resin is an existing conventional biocompatible photocurable resin.

In a preferred embodiment, the cavity table and microneedle array are integrally formed structures.

The utility model also provides a method for preparing the above-mentioned cavity table and microneedle array, and the cavity table and array are prepared by using an integrated molding process.

In a preferred embodiment, the integrated molding process includes but is not limited to 3D printing, micro-casting, template method, and laser etching method.

The paper chip is loaded in the lumen of the connecting tube and on one side close to the hollow microneedle; and/or, all of the paper chips are located in the connecting tube or part of the paper chip is located in the connecting tube and part of the cavity.

In the present invention, the width of the paper chip is smaller than the inner diameter of the connecting tube. Preferably, the width of the paper chip is 70-90% of the inner diameter of the connecting tube. Specifically, the length of the paper chip can be 5-15mm. The inner diameter of the connecting pipe can be 1-10mm.

In the present invention, the paper chip can be a colorimetric test strip, a chemiluminescent test strip, etc., or a lateral chromatography detection test strip, etc.

The paper chip is pre-modified with a chemical sensing system capable of detecting and analyzing tissue fluid components, and its response signal includes color development or self-luminescence. The chemical sensing system can be prepared using existing conventional methods.

Optionally, the paper base of the paper chip can be one of nitrocellulose membrane or filter paper;

Optionally, for the detection functional modification of the paper chip, the entire paper chip substrate can be directly soaked in a solution containing chemical sensing reagents for a period of time, and then taken out to dry.

Optionally, for the detection functional modification of the paper chip, a solution containing chemical sensing reagents can be directly drop-coated on the entire paper chip substrate and then dried.

Optionally, the paper chip can also use a wax spray printer to print a pattern on the paper chip base to form a number of hydrophilic areas as reaction areas and the rest as hydrophobic areas; the hydrophobic areas form hydrophobic blocks between the reaction areas; the paper chip Each hydrophilic region can be modified with different chemical sensing systems for the detection of different components of tissue fluid.

Optionally, when a wax spray printer is used to print a pattern on the paper chip substrate, the hydrophilic areas formed can be located on both front and back sides of the paper chip.

In this utility model, the color or luminescence caused by the interaction between tissue fluid-related components and the chemical sensing system embedded in the paper chip can be collected by the image acquisition device, and then the color data or luminescence intensity of the color can be obtained through image processing software analysis. The data is then compared with the standard system to obtain content information of related components of tissue fluid.

Optionally, the image collection device can be a mobile phone or a camera.

Optional, the image processing software can be Image J, or matlab, or python.

In the utility model, the paper chip reacts with the incoming tissue fluid, and then directly visually reads the T-line and C-line signals of the lateral chromatography test paper to achieve qualitative analysis of the tissue fluid extracted by the hollow microneedle.

Optionally, the lateral chromatography detection test paper chip can be a self-made lateral chromatography detection test paper or a commercially available lateral chromatography detection test paper.

Optional, homemade lateral chromatography detection test paper, consisting of a sample pad, a conjugate release pad, a membrane to fix the capture probe (usually a nitrocellulose membrane), and an adsorption pad, installed on the surface of an inert backing material.

Optionally, the capture probe is one of an antibody and a nucleic acid aptamer.

Optionally, when using commercially available lateral chromatography detection test paper for the lateral chromatography detection test paper chip, you need to take out the lateral chromatography detection test paper from the detection card, then remove the absorbent pad at its rear end and cut it. Shorten the injection area of its front end but leave its sample binding pad intact, and then cut it to size so that it can be inserted into the connecting tube.

Optionally, the lateral chromatography detection test paper can be, but is not limited to, early pregnancy test paper, new coronavirus antigen test paper, hepatitis B virus surface antigen detection test paper, human immunodeficiency virus antibody detection test paper, hepatitis C virus antibody test paper , Treponema pallidum antibody test strips, test strips for detecting illegally added health products and foods taken into the body, test strips for detecting harmful substances in cosmetics taken into the body, and test strips for detecting metal ions taken into the body.

The technical solution of this utility model has the following advantages:

1. The paper chip device provided by this utility model can directly extract tissue fluid from the subcutaneous tissue through the hollow microneedle under the action of the negative pressure generator. The tissue fluid flows from the hollow microneedle into the connecting tube, and the tissue fluid fully contacts the paper chip in the connecting tube. , and then use paper chips to quickly analyze and detect tissue fluids, and the test results can be obtained within 10 minutes.

In addition, the utility model realizes the detection of tissue fluid, and has the advantages of simple operation, on-site detection, rapid detection, and no biological safety risk.

2. The paper chip in the paper chip device provided by the utility model serves as a color development or self-luminous platform and can be directly placed in the lumen of the connecting tube without adding other mechanisms and additional devices. The way to load the paper chip is very simple. Easy. For detection based on chromogenic sensing and lateral chromatography, color changes can be directly observed with the naked eye, and the way to read the detection results is very simple.

3. The chemical sensing system of the paper chip in the paper chip device provided by the present invention has scalability, and can design and realize the detection of a variety of important biomarkers, such as glucose, lactic acid, luteinizing hormone and pH, etc.

Description of drawings

In order to more clearly explain the specific embodiments of the present invention or the technical solutions in the prior art, the drawings that need to be used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings in the following description The accompanying drawings illustrate some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without exerting creative efforts.

Figure 1 is a schematic structural diagram of the paper chip device in Embodiment 1 of the present invention;

Figure 2 is a schematic structural diagram of the paper chip device in Embodiment 2 of the present invention;

Figure 3 is a schematic diagram of the connection relationship between the hollow cavity platform and the hollow microneedle in Embodiment 2 of the present utility model;

Figure 4 is a schematic diagram of the connection relationship between the support body and the cavity in Embodiment 3 of the present invention;

Figure 5 is a schematic structural diagram of the paper chip device in Embodiment 4 of the present invention;

Figure 6 is a schematic diagram of the connection relationship between the support body and the cavity in Embodiment 4 of the present invention;

Figure 7 is a scanning electron microscope image of the hollow microneedle in Example 2 of the present invention;

Figure 8 is a schematic diagram of the verification results of the detection results using the paper chip device of the present invention and a commercially available blood glucose meter in Experimental Example 1;

Reference signs:

1. Cavity table; 2. Hollow microneedle; 3. Paper chip; 4. Connecting tube; 5. Negative pressure generator; 6. Connecting part; 7. Connecting tube lumen; 8. Support body; 9. Storage ; 10. Cavity; 11. Skin tissue.

Detailed ways

The following examples are provided for a better understanding of the present utility model, and are not limited to the best implementation modes. They do not limit the content and protection scope of the present utility model. Anyone inspired by the present utility model or Any product that is identical or similar to the present utility model and is obtained by combining the features of the present utility model with other existing technologies shall fall within the protection scope of the present utility model.

If no specific experimental steps or conditions are specified in the examples, the procedures can be carried out according to the conventional experimental steps or conditions described in literature in the field. If the manufacturer of the reagents or instruments used is not indicated, they are all conventional reagent products that can be purchased commercially.

Example 1

As shown in FIG. 1 , this embodiment provides a paper chip device, including a paper chip 3 , a connecting tube 4 , a hollow microneedle 2 and a negative pressure generator 5 . The paper chip 3 is used for qualitative or quantitative detection of tissue fluid. The paper chip 3 is pre-modified with a chemical sensing system capable of detecting and analyzing tissue fluid components, and its response signal includes color development or luminescence. The paper chip 3 can be homemade according to the method of the present invention, or can be obtained by cutting existing lateral chromatography detection test paper. The paper chip 3 is loaded in the lumen of the connecting tube 4 (that is, the lumen 7 of the connecting tube), and the hollow microneedle 2 is provided with holes. The connecting tube 4 is a through-tube with openings at both ends, and a hollow connecting tube lumen 7 is between the two openings. One end of the connecting tube 4 is connected to the hole on the hollow microneedle 2, and the other end is connected to the negative pressure generator 5 during detection. Under the negative pressure of the negative pressure generator 5, after the hollow microneedle 2 pierces the skin, the tissue fluid flows into the connecting tube lumen 7 along the holes of the hollow microneedle 2, and contacts the paper chip 3 in the connecting tube lumen 7, and the paper Chip 3 performs rapid analysis and instant detection of tissue fluid.

In this embodiment, the paper chip 3 is loaded in the connecting tube lumen 7 and close to the side of the hollow microneedle 2 . The connecting pipe 4 is a transparent pipe. In this embodiment, the connecting tube is a transparent connecting tube used with a disposable venous blood collection needle (specification 0.55×20RW medium purple), and the inner diameter of the lumen is 2.5 mm. The shape of the hollow microneedle 2 is conical. The inner diameter of the pore channel is 300 μm. The length of the hollow microneedle 2 is 800 μm. The paper chip is 1.5mm wide and 10mm long, and can be loaded flatly into the connecting pipe (basically located on the cross section of its central pipe diameter). There is enough space above and below the paper chip, so it does not block the connecting pipe. lumen, thereby leaving enough physical space for liquid flow in the lumen of the connecting tube.

In this embodiment, the negative pressure generator 5 is an existing vacuum sampling tube, that is, a pre-vacuumed sampling tube used when drawing blood in a hospital. The connecting tube 4 is also provided with a hollow tube at one end connected to the negative pressure generator 5 needle. There is a rubber stopper in the center of the top cap of the vacuum sampling tube. During detection, after pressing the hollow microneedle 2 into the subcutaneous tissue, extend the needle end of the connecting tube 4 into the vacuum sampling tube through the rubber stopper. Under the action of negative pressure, The tissue fluid can be directly extracted from the subcutaneous tissue through the hollow microneedle 2. The tissue fluid flows into the connecting tube 4 from the hole of the hollow microneedle 2. The tissue fluid fully contacts the paper chip 3 in the lumen 7 of the connecting tube, and then the paper chip 3 is used to quickly analyze the tissue fluid. and point-of-care testing, with test results available within 10 minutes.

Example 2

As shown in Figures 2 and 3, this embodiment provides a paper chip device. The difference from Embodiment 1 is that this embodiment uses multiple hollow microneedles 2 to form a microneedle array, and the microneedle array is located on the cavity table 1 superior. A cavity 10 structure is formed inside the cavity table 1 . A through-connecting portion 6 is provided on the side of the cavity table 1 . The connecting part 6 has a cylindrical structure, and the connecting pipe 4 is nested with the connecting part 6 . The connecting pipe 4 is a hose and is nested outside the connecting part 6 .

The top surface of the cavity table 1 is square. The bottom surface of the cavity table 1 is a microneedle array. The microneedle array is arranged at the bottom of the cavity table 1 and is structurally connected to the cavity 10. The microneedle array is composed of 5×5 hollow microneedles 2, and each hollow microneedle 2 is provided with 2 self-hollow microneedles. 2 surface penetrates the hole channel on the bottom surface of the cavity table 1. The height of the cavity table 1 is 1 mm; the center distance between the hollow microneedle 2 and adjacent hollow microneedle 2 is 2000 μm; the area of the microneedle array is 1.21cm ² .

The microneedle array and cavity table 1 in this embodiment are integrally formed by 3D printing using biocompatible light-curing resin.

During use, the microneedle array is inserted into the skin tissue 11 by pressing the cavity table 1, and a vacuum sampling tube is connected. Under the action of the negative pressure provided by the vacuum sampling tube, the tissue fluid in the skin tissue 11 flows into the air through the holes of the microneedle array. In the chamber 1, the extracted tissue fluid further enters the vacuum sampling tube through the connecting tube 4 for storage and transportation; after pressing the microneedle array into the skin, it needs to be kept pressed and can be released only after the tissue fluid extraction is completed; after the tissue fluid extraction is completed Finally, first remove the microneedle array from the skin surface, and then separate the connecting tube 4 from the negative pressure generator 5.

Example 3

As shown in Figure 4, this embodiment provides a paper chip device. The difference from Embodiment 2 is that vertical supports 8 are provided in the cavity 10. The number of support bodies 8 is four, surrounding the cavity. The cavities 10 are arranged in the center and are not connected to each other.

Example 4

As shown in Figures 5 and 6, this embodiment provides a paper chip device. The difference from Embodiment 2 is that parallel supports 8 are provided in the cavity 10, and the number of supports 8 is four. On the bottom surface of the cavity table 1, the openings in the cavity table 1 are arranged in parallel avoiding the through holes of the hollow microneedle 2; the negative pressure generator 5 and the reservoir 9 are separate devices for negative pressure generation and storage, and the reservoir 9 It is connected to the connecting tube 4, and the connection point is located between the negative pressure generator 5 and the hollow microneedle 2. The extracted tissue fluid is stored in the reservoir 9, and the negative pressure is provided by the negative pressure generator 5.

Example 5

This embodiment provides a construction method and application of a paper chip device, including:

1. Construction method

(1) Preparation of glucose chromogenic paper chip:

Weigh 0.2g of chitosan, add it to 100.0 mL of 1.0% acetic acid solution, heat, stir and dissolve in an 80°C water bath to obtain a 0.2% (w/v) chitosan acetic acid solution. Cut the nitrocellulose filter paper into a suitable shape and size so that it can be inserted into the lumen of the connecting tube. It was then fully immersed in chitosan acetic acid solution for a period of time and subsequently dried in a 50°C oven. In addition, 10 mg of glucose oxidase, 0.3 mg of horseradish peroxidase and 8 mM 4-aminoantipyrine were dissolved in 1 mL of 0.1 M, pH 6.0 PBS solution. Use a pipette to add the above mixture solution dropwise to the nitrocellulose filter paper pretreated with chitosan acetic acid solution, and then dry it in a 35°C oven for 40 minutes.

(2) Loading: Prepare the connecting tube, vacuum sampling tube, 3D printed integrated microneedle array (patch) and cavity table described in Example 2, and load the constructed glucose chromogenic paper chip into the connecting tube. In the cavity, nest one end of the connecting tube outside the connecting part, and place it in a 4°C refrigerator before use. When in use, connect the other end of the hollow connecting tube to the vacuum sampling tube.

2. Application

Blind samples of glucose solutions of different concentrations were dissolved in 0.1M PBS solution, and agarose powder was added to prepare 1.5% (w/v) agarose hydrogel to simulate human skin tissue.

Press the microneedle array patch into the surface of the hydrogel skin tissue model, maintain the pressed state, and then connect the other end of the hollow connecting tube to the vacuum sampling tube to generate negative pressure, driving the tissue fluid from the hollow microneedle into the connecting tube and contacting the paper chip. Then observe the changes in color reaction of the paper chip in the connecting tube. After the color change remains stable, remove the microneedle array patch device from the skin tissue model. Use a mobile phone as a color reaction image acquisition device, use Matlab software to extract the color of the collected image into R, G, and B values, compare them with the standard system, and calculate the corresponding glucose concentration.

Example 6

This embodiment provides a construction method and application of a paper chip device, including:

1. Construction method

(1) Preparation of multiple chromogenic paper chips:

Using a Xerox Color-Qube 8580Color Printer wax spray printer, print two hydrophilic areas on the same side of the Whatman filter paper. After heating at 90°C for 4 minutes, the paraffin transformed into a molten state on the surface of the filter paper and penetrated the filter paper, forming a hydrophobic block between reaction areas. Dissolve 5mg lactate oxidase, 0.15mg horseradish peroxidase and 10mM o-phenylenediamine in 1mL 0.2% (w/v) chitosan acetic acid solution, and add it dropwise to the hydrophilic area 1 to form a lactic acid detection area . Drop the mixed solution of 0.36mM bromocresol green, 0.46mM bromocresol violet, and 0.08mM bromphenol blue into the hydrophilic area 2 to form a pH detection area.

(2) Loading: Prepare the connecting tube, vacuum sampling tube and single metal hollow microneedle described in Example 1, load the constructed multi-element chromogenic paper chip into the lumen of the connecting tube, and connect one end of the connecting tube to the single metal hollow microneedle. The metal hollow microneedles are connected, and the paper chip partially extends into the hollow microneedle cavity. Refrigerate at 4°C before use.

2. Application

Agarose powder was added to blind samples of different pH to prepare 1.5% (w/v) agarose hydrogel to simulate human skin tissue.

Insert the hollow microneedle into the surface of the hydrogel skin model, and connect the other end of the hollow connecting tube to the vacuum sampling tube. The negative pressure of the vacuum sampling tube drives the tissue fluid from the hollow microneedle into the connecting tube, and observes the changes in the color reaction of the paper chip in the connecting tube. After the color change remains stable, the hollow microneedle paper chip device is removed from the skin tissue. Use SONY A72 camera as the color reaction image acquisition device, use Python software to extract the color of the collected image into R, G, and B values, compare it with the standard system, and calculate the corresponding pH value.

Example 7

This embodiment provides a construction method and application of a paper chip device, including:

1. Construction method

(1) Preparation of multi-element paper chips based on color sensing:

Use a Xerox Color-Qube 8580Color Printer wax spray printer to print two hydrophilic areas on the front and back of the nitrocellulose filter paper. Two hydrophilic areas are arranged staggeredly, respectively at the front end of the front side and the rear end of the back side of the filter paper. After heating at 90°C for 4 minutes, the paraffin melted on the surface of the filter paper and penetrated the filter paper, forming a hydrophobic block between reaction areas. Dissolve 10mg glucose oxidase, 0.3mg horseradish peroxidase and 8mM 4-aminoantipyrine in 1mL of 0.1M, pH 6.0 PBS solution, and add it dropwise to the hydrophilic area 1 on the front end of the filter paper to form Glucose detection area: Dissolve 5mg lactate oxidase, 0.15mg horseradish peroxidase and 10mM o-phenylenediamine in 1mL 0.2% (w/v) chitosan acetic acid solution, and drop it on the back end of the filter paper. Water area 2 forms the lactic acid detection area.

(2) Loading: Prepare the connecting tube, vacuum sampling tube, 3D printed integrated microneedle array (patch) and cavity table described in Example 2, and load the constructed glucose chromogenic paper chip into the connecting tube. In the cavity, nest one end of the connecting tube outside the connecting part, and place it in a 4°C refrigerator before use. When in use, connect the other end of the hollow connecting tube to the vacuum sampling tube.

Example 8

This embodiment provides a construction method and application of a paper chip device, including:

1. Construction method

(1) Preparation of glucose paper chip based on self-luminescence sensing:

Dip the filter paper into a buffer solution containing 20mM Co ²⁺ , 0.8mM dimethylimidazole, 2mg/mL glucose oxidase, 15mM luminol and 50mM HEPES, then place it on a shaker at 30°C for 20 minutes, and then dry under vacuum.

(2) Loading: Prepare the connecting tube, vacuum sampling tube, 3D printed integrated microneedle array (patch) and cavity table described in Example 2, and load the constructed glucose chromogenic paper chip into the connecting tube. In the cavity, nest one end of the connecting tube outside the connecting part, and place it in a 4°C refrigerator before use. When in use, connect the other end of the hollow connecting tube to the vacuum sampling tube.

2. Application

Blind samples of glucose solutions of different concentrations were dissolved in 0.1M PBS solution, and agarose powder was added to prepare 1.5% (w/v) agarose hydrogel to simulate human skin tissue.

Press the microneedle array patch into the surface of the hydrogel skin tissue model, maintain the pressed state, and then connect the other end of the hollow connecting tube to the vacuum sampling tube to generate negative pressure, driving the tissue fluid from the hollow microneedle into the connecting tube and contacting the paper chip. Then observe the changes in the self-luminescence reaction of the paper chip in the connecting tube, and use a mobile phone as a luminous intensity collection device to take pictures, with an exposure of 60 seconds. The captured pictures are converted into grayscale values using ImageJ software, compared with the standard system, and the corresponding calculation is obtained glucose concentration.

Example 9

This embodiment provides an application of a paper chip device, including:

Blind samples of luteinizing hormone solutions of different concentrations were dissolved in 0.1M PBS solution, and agarose powder was added to prepare 1.5% (w/v) agarose hydrogel to simulate human skin.

Prepare the connecting tube, vacuum sampling tube, 3D printed integrated microneedle array (patch) and cavity table described in Example 2, and cut the commercially available luteinizing hormone test strip (paper chip) (1.5 mm wide, 10mm long) and then load it into the lumen of the connecting tube, and nest one end of the connecting tube outside the connecting part. Press the microneedle array patch into the surface of the hydrogel skin tissue model, maintain the pressed state, and then connect the other end of the hollow connecting tube to the vacuum sampling tube to generate negative pressure, driving the tissue fluid from the hollow microneedle into the connecting tube and contacting the paper chip. Then observe with the naked eye the color development of the C-line and T-line of the paper chip in the hollow connecting tube. After the color change remains stable, the luteinizing hormone test result is obtained.

Experimental Example 1 Color Detection and Verification of Glucose in Skin Model

Blind samples of glucose solutions of different concentrations were dissolved in 0.1M PBS solution, and agarose powder was added to prepare 8 groups of 1.5% (w/v) agarose hydrogels with different glucose concentrations to simulate human skin tissue. The test group and control group methods were used for detection respectively.

Test group: The paper chip device and application method of Example 5 of the present invention are used to detect the agarose hydrogel simulated human skin tissue, and calculate the glucose content in the blind sample of the glucose solution.

Control group: Press the agarose hydrogel to simulate human skin tissue, wait for the liquid to flow out, and use a commercially available blood glucose detector to detect the liquid.

The results of the test group and the control group were compared for similarity. The result is shown in Figure 7. R ² =0.9990, which proves that the paper chip device color sensing system used in the present utility model has high accuracy.

Experimental Example 2 Paper chip device for rapid on-site detection of glucose content in living tissue fluid of New Zealand rabbits

Weigh a New Zealand rabbit (2.5kg, female) and fix it on a rabbit fixed table. Prepare sodium pentobarbital physiological saline solution at a dose of 1mL/kg and 3%. Inject sodium pentobarbital slowly along the ear edge vein. Anesthetic. After confirming successful anesthesia, disinfect with iodophor. Use a depilator to remove hair on the back of the rabbit's ears and wipe clean with saline.

The paper chip device and application method of Embodiment 5 of the present invention are used to extract and detect tissue fluid from rabbit ear skin. Use a mobile phone as a color reaction image acquisition device, and use Matlab software to extract the color of the collected image into R, G, and B values. Compare it with the standard system and calculate the corresponding glucose concentration to be 5.99mmol/L. After the detection of the paper chip device in Embodiment 5 of the present invention is completed, remove the connecting tube attached to the hollow microneedle, and attach the connecting part of the hollow microneedle to the test card inlet of the commercially available blood glucose meter, so that the hollow microneedle The remaining tissue fluid in the needle patch enters the blood glucose meter detection card through the connection part, and the glucose concentration is detected to be 5.99mmol/L. Another rabbit ear margin vein blood was taken, and a commercially available blood glucose detector was used to measure the glucose concentration of the rabbit ear margin vein blood to be 6.00mmol/L, confirming that the paper chip color sensing system used in the present utility model has high accuracy.

Experimental Example 3 Paper chip device for rapid on-site detection of pH content of New Zealand rabbit living tissue fluid

Weigh a New Zealand rabbit (2.5kg, female) and fix it on a rabbit fixed table. Prepare sodium pentobarbital physiological saline solution at a dose of 1mL/kg and 3%. Inject sodium pentobarbital slowly along the ear edge vein. Anesthetic. After confirming successful anesthesia, disinfect with iodophor. Use a depilator to remove hair on the back of the rabbit's ears and wipe clean with saline.

The paper chip device and application method of Embodiment 6 of the present invention are used to extract tissue fluid from rabbit ear skin and perform detection. A SONY A72 camera was used as the color reaction image acquisition device, and Python software was used to extract the color of the collected image into R, G, and B values. Compared with the standard system, the corresponding pH value was calculated to be 7.40. Since the pH value of the healthy rabbit ear subcutaneous tissue fluid is similar to that of the rabbit ear marginal vein blood, the rabbit ear marginal vein blood was taken and the pH of the New Zealand rabbit ear marginal vein blood was measured using a laboratory standard pH meter to be 7.40, confirming that the paper used in the present utility model The chip color sensing system has high accuracy.

Obviously, the above-mentioned embodiments are only examples for clear explanation and are not intended to limit the implementation. For those of ordinary skill in the art, other different forms of changes or modifications can be made based on the above description. An exhaustive list of all implementations is neither necessary nor possible. The obvious changes or modifications derived therefrom are still within the protection scope of the invention.

Claims (8)

1. The paper chip device is characterized by comprising a connecting pipe and a negative pressure generator, wherein a paper chip is arranged in a lumen of the connecting pipe, one end of the connecting pipe is communicated with a hollow microneedle, the other end of the connecting pipe is communicated with the negative pressure generator during detection, and a pore canal communicated with the connecting pipe is arranged on the hollow microneedle.

2. The paper chip device according to claim 1, wherein a cavity stage is further provided between the connection pipe and the hollow microneedle, a cavity structure is formed inside the cavity stage, a cavity of the cavity stage is communicated with a duct of the hollow microneedle, and the duct of the hollow microneedle is communicated with the connection pipe through the cavity stage.

3. The paper chip device according to claim 2, wherein a connecting portion communicating with a connecting pipe is provided at one side of the cavity stage, and a cavity of the cavity stage communicates with the connecting pipe through the connecting portion.

4. The paper chip device of claim 1, wherein a plurality of tunnels are provided on the hollow microneedles; and/or, the hollow microneedles are arranged in a plurality to form a microneedle array.

5. The paper chip device of claim 2, wherein a support body is disposed within the cavity that connects the top and bottom of the cavity table.

6. The paper chip device of claim 5, wherein the support is disposed away from the opening of the hollow microneedle; and/or the supporting bodies are arranged in parallel with each other or around the center of the cavity; and/or the number of the supporting bodies is 2-4; and/or the support body is provided with a through hole; and/or the supporting bodies are not connected with each other; and/or the support body is not connected with the inner wall in the cavity.

7. The paper chip device of claim 1, wherein the paper chip is loaded in the connecting tube lumen and adjacent to one side of the hollow microneedle; and/or the paper chip is wholly located in the connecting pipe or partly located in the connecting pipe and partly located in the cavity.

8. The paper chip device of claim 1, wherein the paper chip is pre-modified with a chemical sensing system capable of detecting and analyzing tissue fluid components, the response signal of which comprises color development or luminescence.