CN212674775U - Micro-fluidic chip detection system - Google Patents
Micro-fluidic chip detection system Download PDFInfo
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- CN212674775U CN212674775U CN202020737626.1U CN202020737626U CN212674775U CN 212674775 U CN212674775 U CN 212674775U CN 202020737626 U CN202020737626 U CN 202020737626U CN 212674775 U CN212674775 U CN 212674775U
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Abstract
The utility model provides a micro-fluidic chip detecting system relates to immunology technical field, micro-fluidic chip detecting system includes light source, micro-fluidic chip and image acquisition equipment, the light source is used for shining micro-fluidic chip, image acquisition equipment is used for gathering under the light source shines the image of detecting sample among the micro-fluidic chip. The utility model provides a micro-fluidic chip detecting system adopts the light source to shine micro-fluidic chip to make image acquisition system can gather the image of detecting sample among the micro-fluidic chip, through the image of determinand among the micro-fluidic chip that analysis image acquisition system gathered, obtain the volume of the determinand of participating in the reaction, convenient and fast.
Description
Technical Field
The utility model belongs to the technical field of the immunology technique and specifically relates to a micro-fluidic chip detecting system is related to.
Background
The micro-fluidic chip technology integrates basic operation units of sample preparation, reaction, separation, detection and the like in the biological, chemical and medical analysis process into a micron-scale chip, and automatically completes the whole analysis process. Due to its great potential in the fields of biology, chemistry, medicine and the like, the method has been developed into a new research field crossing the disciplines of biology, chemistry, medicine, fluid, electronics, materials, machinery and the like.
With the development of microfluidic technology, more and more microfluidic chips are applied in the field of clinical detection. The precision and repeatability of the structure of the microfluidic chip ensure that the difference between batches of the microfluidic chip is small, and the result can be repeated; the micro-channel ensures that the required sample amount is very small, only micro-liter level samples are needed, and the method is convenient and quick. However, the current microfluidic chip in the field of clinical detection mainly obtains the concentration of the analyte by detecting the fluorescence intensity after labeling fluorescein and a detection antigen or antibody, and cannot determine the volume of the analyte participating in the reaction.
In view of this, the present invention is especially provided.
SUMMERY OF THE UTILITY MODEL
An object of the utility model is to provide a micro-fluidic chip detecting system to detect the image of sample in the micro-fluidic chip through analysis micro-fluidic chip detecting system collection, obtain the volume of the determinand of participating in the reaction.
The utility model provides a micro-fluidic chip detecting system, including light source, micro-fluidic chip and image acquisition equipment, the light source is used for shining micro-fluidic chip, image acquisition equipment is used for gathering under the light source shines the image of detecting the sample among the micro-fluidic chip.
Furthermore, the microfluidic chip comprises a sample adding area, a combining area, a reaction area and a waste liquid area which are sequentially communicated.
Furthermore, in the microfluidic chip, a filtering area is arranged between the sample addition area and the combining area.
Furthermore, one end of the binding region is conical and is used for being communicated with the sample adding region, and the other end of the binding substance is in a micro-channel shape and is used for being communicated with the reaction region.
Furthermore, in the microfluidic chip, the reaction area comprises a micro-pipeline, and the diameter of the micro-pipeline is 10 μm-4 mm.
Further, the image acquisition device comprises at least one of a camera, a video camera and a video camera.
Further, the light source is a monochromatic light source.
Further, a first lens and a first color filter are sequentially arranged between the light source and the microfluidic chip.
Furthermore, the microfluidic chip detection system further comprises a fluorescence detector, and the fluorescence detector is used for determining the fluorescence intensity of a detection sample in the microfluidic chip.
Furthermore, a second lens and a second color filter are sequentially arranged between the fluorescence detector and the microfluidic chip.
The utility model provides a micro-fluidic chip detecting system adopts the light source to shine micro-fluidic chip to make image acquisition system can gather the image of detecting sample among the micro-fluidic chip, through the image of determinand among the micro-fluidic chip that analysis image acquisition system gathered, obtain the volume of the determinand of participating in the reaction, convenient and fast.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the embodiments or the technical solutions in the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.
Fig. 1 is a schematic structural diagram of a microfluidic chip detection provided in embodiment 1 of the present invention;
fig. 2 is a schematic structural diagram of a microfluidic chip provided in embodiment 1 of the present invention;
fig. 3 is a schematic structural diagram of a microfluidic chip detection system provided in embodiment 2 of the present invention.
Icon:
100-a microfluidic chip; 101-a sample addition zone; 102-a filtration zone; 103-a binding domain; 104-a reaction zone; 105-a waste liquid zone; 201-a light source; 202-a first lens; 203-a first color filter; 300-an image acquisition device; 401-fluorescence detector; 402-a second lens; 403-second color filter.
Detailed Description
The technical solution of the present invention will be described clearly and completely with reference to the accompanying drawings, and obviously, the described embodiments are some, but not all embodiments of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative work belong to the protection scope of the present invention.
In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, and are only for convenience of description and simplification of description, but do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus, should not be construed as limiting the present invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
In the description of the present invention, it is to be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meaning of the above terms in the present invention can be understood in specific cases to those skilled in the art.
Most of the existing microfluidic detection kits in the in vitro diagnostic product market utilize the principle of antigen and antibody specificity combination, use different fluorescein as a labeled antibody or antigen, and form a complex with the antibody or antigen of a stationary phase and the specific antigen or antibody in a sample to be detected, and finally detect the fluorescence intensity of the complex to complete the determination of the concentration of the substance to be detected in the sample. There is no apparatus for determining the volume of the analyte.
According to the utility model discloses a first aspect, the utility model provides a micro-fluidic chip detecting system, including light source, micro-fluidic chip and image acquisition equipment, the light source is used for shining micro-fluidic chip, image acquisition device is used for gathering under the light source shines the image of detecting sample among the micro-fluidic chip.
The utility model discloses in, the light source is used for shining the micro-fluidic chip, and the light source can be monochromatic source, and wherein, monochromatic source can but is not limited to for monochromatic LED light source.
The utility model discloses an in a preferred embodiment, the central wavelength signal of monochromatic LED light source is greater than fluorescence signal's wavelength, still can be for ruddiness or near infrared light, and then adopts the utility model discloses a light source can reduce and cause the interference when carrying out fluorescence detection.
The utility model discloses an in the preferred embodiment, first lens and first color filter have set gradually between light source and the micro-fluidic chip to further improve the interference killing feature of light source, reduce the divergence angle and improve the illuminating effect.
In a preferred embodiment of the present invention, the microfluidic chip comprises a sample addition region, a binding region, a reaction region and a waste liquid region which are sequentially connected.
The utility model discloses an among the preferred embodiment, the application of sample district is used for dropwise add the determinand, and the determinand passes through the application of sample district and gets into the complex that combines back of first antigen/antibody and fluorescence microballon in bonding region and the bonding region under capillary action, continues to flow into the reaction zone, combines with the second antigen/antibody of fixing in the reaction zone bottom at the reaction zone, and the determinand that does not combine with second antigen/antibody in the reaction zone flows into waste liquid district.
In a preferred embodiment of the present invention, the sample adding region is configured as a circular or oval cavity structure, and may be configured as other shapes.
In a preferred embodiment of the present invention, the binding region is disposed between the sample adding region and the reaction region, one end of the binding region is tapered for communicating with the sample adding region, and the other end is in the shape of a micro-channel for communicating with the reaction region.
The utility model discloses an among the preferred embodiment, be provided with the filtering area between appearance district and the combination district with adding to do benefit to and get rid of the impurity in the determinand, avoid the existence of impurity to influence the testing result.
In a further preferred embodiment of the present invention, the filtration zone may be composed of microcolumns arranged in an array, the diameter of the microcolumns is 1 μm to 2mm, and the distance between adjacent microcolumns is 10 μm to 5 mm.
Typically, but not limited to, the diameter of the microcolumn is, for example, 1 μm, 2 μm, 5 μm, 8 μm, 10 μm, 12 μm, 15 μm, 18 μm, 20 μm, 30 μm, 50 μm, 80 μm, 1mm, 1.2mm, 1.5mm, 1.8mm or 2 mm.
Typically, but not by way of limitation, the distance between adjacent microcolumns is 10 μm, 12 μm, 15 μm, 18 μm, 20 μm, 30 μm, 50 μm, 80 μm, 1mm, 1.5mm, 2mm, 2.5mm, 3mm, 3.5mm, 4mm, 4.5mm, or 5 mm.
In an embodiment of the present invention, the filter area is composed of filter paper, and the filter paper selects different models according to the difference of the object to be measured.
In a preferred embodiment of the present invention, the reaction zone comprises microchannels, the diameter of which is between 10 μm and 2 mm.
Typically, but not by way of limitation, the diameter of the microchannels may be, for example, 10 μm, 12 μm, 15 μm, 18 μm, 20 μm, 30 μm, 50 μm, 80 μm, 1mm, 1.2mm, 1.5mm, 1.8mm or 2 mm.
In one embodiment of the present invention, the reaction area is in the shape of a micro-pipe, so that the analyte can flow in the reaction area under the action of the capillary.
In a preferred embodiment of the present invention, the image capturing device includes at least one of a camera, a video camera, and a still camera.
The above-mentioned "at least one" means that the image acquisition device may be a camera, a video camera or a camera, or two or three of the camera, the video camera or the camera may be adopted to acquire image information at the same time.
In a preferred embodiment of the present invention, the acquired image information includes, but is not limited to, pictures or videos.
In a preferred embodiment of the present invention, the micro-fluidic chip detection system can further include a fluorescence detector for detecting the fluorescence intensity of the reaction region, so as to calculate the concentration of the analyte according to the fluorescence intensity.
In a preferred embodiment of the present invention, a second lens and a second color filter are sequentially disposed between the fluorescence detector and the micro-fluidic chip to further reduce interference and improve the accuracy of detection.
According to the second aspect of the utility model, the utility model provides an above-mentioned micro-fluidic chip detecting system is in the application of antigen antibody immunity detection.
Through adopting the utility model discloses the micro-fluidic detecting system that the first aspect provided can be under the condition that need not set for quantitative reaction chamber can convenient and fast survey the volume of the determinand of participating in the reaction, effectively reduced antigen-antibody immunity detection's cost, have wide application prospect.
According to the utility model discloses a third aspect, the utility model provides an antigen antibody immunodetection method, it adopts the utility model discloses the micro-fluidic chip detecting system that the first aspect provided can be on the basis of not reforming transform current micro-fluidic chip structure, through the image that acquires the determinand when the micro-fluidic chip of flow-through, obtains the volume of the determinand of participating in the reaction, has effectively reduced antigen antibody immunodetection's cost, has wide application prospect.
In the present invention, the "analyte participating in the reaction" refers to an analyte capable of reacting with the second antigen/antibody immobilized in the binding region.
In a preferred embodiment of the present invention, the method for immunodetection of antigen and antibody comprises the following steps:
(a) arranging a first antigen/antibody and tracer compound at the bottom of the binding region, and fixing a second antigen/antibody at the bottom of the reaction region;
(b) adding the substance to be detected into the sample adding area, wherein the substance to be detected flows into the binding area through the sample adding area, flows into the reaction area after the binding area is combined with the first antigen/antibody and the tracer compound, and is combined with the second antigen/antibody in the reaction area, and the substance to be detected which is not combined with the second antigen/antibody flows into the waste liquid area;
(c) and respectively acquiring images of the material flow to be detected passing through the combination area and the reaction area by adopting an image acquisition device, and analyzing the images of the material to be detected in the combination area and the reaction area to obtain the volume of the material to be detected participating in the reaction.
In a preferred embodiment of the present invention, in step (a), the first antigen/antibody and tracer complex disposed at the bottom of the reaction zone is capable of flowing freely within the reaction zone.
In a preferred embodiment of the present invention, "/" represents "or", the above-mentioned "first antigen/antibody" means "first antigen or first antibody", and the above-mentioned "second antigen/antibody" means "second antigen or second antibody".
In a preferred embodiment of the present invention, the tracer includes, but is not limited to, fluorescent microspheres and enzyme conjugates.
In one embodiment of the present invention, in step (a), the first antigen/antibody and fluorescent microsphere complex disposed at the bottom of the reaction region is prepared by the following method:
(1) taking a rare earth fluorescent microsphere solution, activating, adding a first antibody/antigen for marking, carrying out sealing treatment on microspheres marked with the first antigen/antibody, and then cleaning and re-suspending to obtain a first antigen/antibody and fluorescent microsphere compound;
(2) and spotting the first antigen/antibody and fluorescent microsphere compound to a binding region on the microfluidic chip, and drying to finish the arrangement of the first antigen/antibody and fluorescent microsphere compound at the bottom of the reaction region.
In a preferred embodiment of the present invention, the fluorescent microsphere is a fluorescent microsphere containing rare earth element Eu or Tb or Sm, and the fluorescent microsphere is formed by polymerizing one or more of polystyrene, silica, ferroferric oxide, and other materials.
In a preferred embodiment of the present invention, the surface of the fluorescent microspheres includes, but is not limited to, microspheres with or without carboxyl, amino modifications.
In a preferred embodiment of the present invention, the fluorescent microspheres have a diameter of 10nm to 100 μm.
Typically, but not by way of limitation, the rare earth fluorescent microspheres have a diameter of, for example, 10 μm, 20 μm, 50 μm, 80 μm, 1 μm, 2 μm, 5 μm, 8 μm, 10 μm, 20 μm, 30 μm, 50 μm, 70 μm, 80 μm, 90 μm, or 100 μm.
In a preferred embodiment of the present invention, the activation of the fluorescent microspheres includes, but is not limited to, activation of the fluorescent microspheres with EDC, glutaraldehyde, and the like.
In a preferred embodiment of the present invention, the complex of the first antibody/antigen and the fluorescent microsphere is a complex that labels one or more antibodies or antigens simultaneously.
In a preferred embodiment of the present invention, the drying after spotting the binding domains is performed by methods including, but not limited to, oven drying, freeze drying, and vacuum drying.
In a preferred embodiment of the present invention, in step (a), the second antigen/antibody immobilized at the bottom of the binding region is prepared by the following method:
spotting the second antigen/antibody on the reaction zone so that the second antigen/antibody is immobilized on the bottom of the reaction zone, and drying to complete the immobilization of the second antigen/antibody on the bottom of the binding zone.
In a preferred embodiment of the present invention, the drying method includes, but is not limited to, natural air drying, oven drying, freeze drying, vacuum drying, and the like.
In a preferred embodiment of the present invention, the second antigen/antibody is spotted on the reaction zone, and the spotting and immobilization method includes, but is not limited to, passive adsorption, covalent coupling, nucleophilic adsorption, and the like.
In a preferred embodiment of the present invention, one or more antibodies/antigens can be labeled on the microfluidic chip in a quantitative manner, so that a plurality of items of a single sample can be detected simultaneously, and the detection efficiency can be improved.
In a preferred embodiment of the present invention, in step (c), the image P after the analyte enters the binding region and fully binds to the first antigen/antibody and the fluorescent microsphere complex is collected by the image collecting device1And an image P of a conjugate of the substance to be detected and the first antigen/antibody bound to the fluorescent microsphere complex, which flows through the binding region, enters the reaction region, and then is fully bound to the second antigen antibody2Through P1And P2Respectively calculating to obtain the volume V of the object to be measured flowing through the binding area and the reaction area1And V2And calculating the difference value of the two values to obtain the volume of the object to be detected participating in the reaction.
In a preferred embodiment of the present invention, the image P is used1And P2Calculating V1And V2An example of the algorithm is as follows:
since the width and height of the micro-channels of the binding region and the reaction region in the microfluidic chip are known, taking the image P as an example of the micro-channel with the binding region of the first antigen/antibody and the tracer as a 2mm wide micro-channel1Counting the number of pixels in a square with the upper side length of the upper microchannel equal to the width of the microchannel, thereby obtaining the corresponding relation between the area and the number of the pixels, and further obtaining the corresponding relation according to the shot image P1The total image area is obtained by calculating the number of pixel points in the image, and the volume V of the object to be measured flowing through the binding area can be calculated by multiplying the total image area by the height of the micro-channel1Similarly, the volume V of the analyte flowing through the reaction region can be calculated2。
The utility model discloses an in a preferred embodiment, when micro-fluidic chip detecting system was provided with fluorescence detector, can also calculate the concentration that obtains the determinand according to the fluorescence intensity of the reaction zone that fluorescence detector detected.
In a preferred embodiment of the present invention, the analyte includes at least one of a small molecule substance, an antigen, an antibody, a hormone, an antibiotic, a bacterium, and a virus.
The term "at least one" as used herein means that the analyte may be any one of a small molecule substance, an antigen, an antibody, a hormone, an antibiotic, a bacterium or a virus, or a mixture of two or more of the above substances.
In a further preferred embodiment of the present invention, the small molecule substance includes, but is not limited to, small molecule drugs, and the like.
The technical solution provided by the present invention will be further described with reference to the following embodiments.
Example 1
The embodiment provides a microfluidic chip detection system, as shown in fig. 1, including a light source 201, a microfluidic chip 100, and an image acquisition device 300, where the light source 201 is used to irradiate the microfluidic chip 100, and the image acquisition device 300 is used to acquire an image of a detection sample in the microfluidic chip 100 under the irradiation of the light source 201.
In a preferred embodiment of this embodiment, a first lens 202 and a first color filter 203 are sequentially disposed between the light source 201 and the microfluidic chip 100, so as to further improve the interference resistance of the light source 201, reduce the divergence angle, and improve the illumination effect.
In a preferred embodiment of this embodiment, as shown in fig. 2, the microfluidic chip 100 includes a sample addition region 101, a binding region 103, a reaction region 104 and a waste liquid region 105, which are connected in sequence.
In a preferred embodiment of this embodiment, the sample addition region 101 is used for dropping a sample, the sample enters the binding region 103 through the sample addition region 101 under the action of capillary, and is combined with the complex of the first antigen/antibody and the fluorescent microsphere in the binding region 103, and then continuously flows into the reaction region 104, the second antigen/antibody fixed at the bottom of the reaction region 104 is combined in the reaction region 104, and the sample which is not combined with the second antigen/antibody in the reaction region 104 flows into the waste liquid region 105.
In a preferred embodiment of this embodiment, the reaction zone 104 comprises microchannels having a diameter of 10 μm to 4 mm.
In a preferred embodiment of this embodiment, the image capturing device 300 is selected from one of a camera, a video camera, or a still camera.
In a preferred embodiment of this embodiment, a filtering region 102 is disposed between the sample-adding region 101 and the binding region 103, so as to facilitate removing impurities in the analyte and avoid the presence of impurities from affecting the detection result.
Example 2
Fig. 3 is a schematic structural diagram of a microfluidic chip detection system provided in embodiment 2 of the present invention; the present embodiment is an improvement on embodiment 1, and is different from embodiment 1 in that the microfluidic chip detection system provided in the present embodiment is further provided with a fluorescence detector 401.
In the embodiment, the fluorescence detector 401 is arranged in the microfluidic chip detection system, so that the fluorescence intensity of the reaction region 104 is detected by the fluorescence detector 401, and the concentration of the analyte is calculated according to the fluorescence intensity.
In a preferred embodiment of this embodiment, a second lens 402 and a second color filter 403 are sequentially disposed between the fluorescence detector 401 and the microfluidic chip 100, so as to further reduce interference and improve detection accuracy.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; although the present invention has been described in detail with reference to the foregoing embodiments, it should be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; such modifications and substitutions do not depart from the spirit and scope of the present invention.
Claims (8)
1. The microfluidic chip detection system is characterized by comprising a light source, a microfluidic chip and an image acquisition device, wherein the light source is used for irradiating the microfluidic chip, and the image acquisition device is used for acquiring an image of a detection sample in the microfluidic chip under the irradiation of the light source; the microfluidic chip comprises a sample adding area, a combining area, a reaction area and a waste liquid area which are sequentially communicated;
a filtering area is arranged between the sample adding area and the combining area;
the filtering area is composed of micro-columns which are arranged in an array, the diameter of each micro-column is 1 mu m-2 mm, and the distance between every two adjacent micro-columns is 10 mu m-5 mm.
2. The microfluidic chip detection system according to claim 1, wherein one end of the binding region is tapered for communicating with the sample application region, and the other end of the binding region is in the shape of a microchannel for communicating with the reaction region.
3. The microfluidic chip detection system according to claim 1, wherein the reaction region comprises a micro channel having a diameter of 10 μm to 4 mm.
4. The microfluidic chip detection system according to claim 1, wherein the image capturing device is a camera, a video camera or a still camera.
5. The microfluidic chip detection system according to claim 1, wherein the light source is a monochromatic light source.
6. The microfluidic chip detection system according to claim 1, wherein a first lens and a first color filter are sequentially disposed between the light source and the microfluidic chip.
7. The microfluidic chip detection system according to any one of claims 1 to 6, further comprising a fluorescence detector for determining fluorescence intensity of a sample detected in the microfluidic chip.
8. The microfluidic chip detection system according to claim 7, wherein a second lens and a second color filter are sequentially disposed between the fluorescence detector and the microfluidic chip.
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