CN209940977U - Cell suction printing enrichment and suction printing dyeing integrated reaction device - Google Patents
Cell suction printing enrichment and suction printing dyeing integrated reaction device Download PDFInfo
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- CN209940977U CN209940977U CN201920564841.3U CN201920564841U CN209940977U CN 209940977 U CN209940977 U CN 209940977U CN 201920564841 U CN201920564841 U CN 201920564841U CN 209940977 U CN209940977 U CN 209940977U
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Abstract
The utility model discloses a cell absorbing, printing, enriching and absorbing, printing and dyeing integrated reaction device, which comprises a microporous membrane (1), wherein the microporous membrane (1) is provided with a fixing piece (2), and the fixing piece (2) is provided with a reaction tank (6); an upper cover (7) is arranged on the reaction tank (6); a liftable objective table (3) is arranged on one side of the fixing piece (2), and a storage bin (5) is arranged below the fixing piece (2); the storage bin (5) is filled with water-absorbing substances. When the cell suspension passes through the microporous membrane, cells with the pore size larger than that of the membrane are intercepted in the reaction container and adsorbed on the membrane surface due to the adoption of an attraction filtration interception mode. The water-absorbing material has stable absorption speed, soft absorption force, slow speed and no pressure, so that the cells are attached to the surface of the membrane.
Description
Technical Field
The utility model belongs to the field of medical equipment, concretely relates to cell is inhaled and is printed enrichment and inhale and print dyeing integration reaction unit, the absorption of nucleated cell (malignant tumor cell, blood disease cell, cardiovascular cast-off cell, parent blood fetal cell and microbial cell) etc. among the mainly used body fluid (blood, urine, ascites in chest, cerebrospinal fluid, joint effusion, saliva, sputum, secretion, throat and cervical swab etc.) is printed enrichment and is inhaled and is printed the dyeing appraisal, has important clinical application and worth.
Background
At present, nucleated pathological cells are enriched from body fluids, wherein the main means are as follows: (1) CTC (peripheral blood tumor cell) dependent antibody based capture of malignant cells in enriched blood: more false positives or false negatives may occur due to differences or deletions in the expression of malignant cell antigens, poor capture efficiency and large variability (sometimes quite low); (2) the CTC is enriched by adopting an ISET type cell filtration and interception technology depending on the cell size, and because positive pressure and negative pressure are adopted to drive or attract cells in liquid to pass through the microporous membrane, the cells are passively accumulated and accumulated on the surface of the microporous membrane by pressure or attraction, the morphological structure of the cells can be changed or destroyed, or the microporous membrane is blocked, or the microporous membrane is penetrated into waste liquid, so that inaccurate dyeing identification or omission occurs, and false positive or false negative occurs; (3) examination of exfoliated cells: few target cells and many mixed cells are collected, and the pathological cells appearing in body fluid are difficult to accurately and sensitively detect and identify.
SUMMERY OF THE UTILITY MODEL
The utility model aims to provide a reaction device integrating cell absorption printing enrichment and absorption printing dyeing, which is based on a microporous membrane capillary microfiltration reaction method and a reaction device, does not depend on antibodies, and enriches and intercepts nucleated pathological cells in body fluid through the microporous membrane capillary microfiltration device; and carrying out cytochemistry, cytoimmunology and cytogenetics reactions on the microporous membrane enriching and intercepting the nucleated pathological cells, and absorbing reacted liquid or washing liquid through a capillary microfiltration device to finish the procedures of enriching and identifying the cells. And (3) absorbing, printing and enriching malignant cells, cardiovascular cast-off cells, maternal blood fetal cells and microbial cells appearing in body fluid.
The cell absorbing, printing, enriching and absorbing, printing and dyeing integrated reaction device comprises a microporous membrane, wherein the microporous membrane is provided with a fixing piece, and a reaction groove is arranged on the fixing piece; an upper cover is arranged on the reaction tank; a liftable object stage is arranged on one side of the fixing piece, and a storage bin is arranged below the fixing piece; the storage bin is filled with water-absorbing substances.
The microporous membrane is specifically a polycarbonate membrane, a polyether sulfone membrane, a nitrocellulose membrane and the like. In a preferred embodiment, the microporous membrane may also be provided with a rigid membrane support, such as a silicon mesh, a metal mesh, or the like.
The microporous membrane is connected with the fixing piece; the fixing piece is of a common glass sheet type: making holes on two glass sheets, adhering or clamping the microporous membrane between the holes, and supporting and clamping the microporous membrane by using an annular pad (preventing cells and reaction liquid from leaking); one of the slide is bonded with a reaction tank to be filled with liquid for adding body fluid or reaction liquid.
The microporous membrane is connected with the fixing piece; the mounting print the glass formula for 3D: the reaction tank is directly printed above the holes, the microporous membrane is adhered or clamped between the two holes of the upper and lower sheets, and the microporous membrane is supported and clamped by a ring-shaped pad.
The inner diameter of the reaction tank is equal to the diameter of the microporous membrane.
The outer diameter of the reaction tank is equal to the inner diameter of the upper cover.
The ratio of the height of the reaction tank to the height of the upper cover is 1: (1.1-1.2).
The ratio of the inner diameter of the storage bin to the inner diameter of the reaction tank is (1.5-1.6): 1.
the fixing piece is connected with the liftable object stage through the socket.
The absorption printing and enrichment and absorption printing and dyeing integrated reaction device for the nucleated cells in the body fluid further comprises a base, and the liftable objective table is connected with the base; the storage bin is arranged in the base.
The microporous membrane, the reaction tank and the storage bin are coaxially arranged from top to bottom.
The water-absorbing substance comprises water-absorbing paper (including filter paper, sanitary towel, toilet paper, etc.), water-absorbing sponge, water-absorbing resin or water-absorbing cotton, etc.
Compared with the prior art, the beneficial effects of the utility model are that:
the detection of these pathogenic microorganisms from body fluids is currently mainly carried out by culture and smear immunology methods, wherein the culture and subsequent identification time is long, the operation is complicated, and the detection is especially not suitable for emergency treatment and ICU (intensive care unit) patient specimen examination. The use of antibiotics, etc., can also result in pathogen variability that is difficult to detect. The smear detection sensitivity is low. The immunological detection is generally to detect antibodies against pathogens (the amount of antigens is relatively low and difficult to detect), and early detection and early diagnosis are difficult.
The device is actually an integrated laboratory for enriching and dyeing nucleated cells, a water absorbing substance part absorbs, receives and drives sample liquid and reaction liquid to flow, the reaction liquid covers and suspends the cell liquid, a microporous membrane filters and intercepts target objects (malignant cells and pathogenic organisms), and the water absorbing substance absorbs and drives the sample and the reaction liquid to complete all reactions on the microporous membrane.
(1) Uptake of trapped material based on water-absorbing material: when the cell suspension passes through the microporous membrane, cells with the pore size larger than that of the membrane are intercepted in the reaction container and adsorbed on the membrane surface due to the adoption of an attraction filtration interception mode. The water-absorbing material has stable absorption speed, soft absorption force, slow speed and no pressure, so that the cells are attached to the surface of the membrane.
(2) Low-speed filtration and interception: the shearing force for intercepting the cells by low-speed filtration is small, and the morphological structure of the cells is not damaged.
(3) The blotting reaction time can be set: the reactant (antigen antibody, marker, etc.) is contacted with the cell sufficiently and reacted for a certain time, so as to achieve the purpose of fully reacting and fully increasing the detection sensitivity.
(4) The washing time and the number of washing times can be set: sufficient washing can remove non-specific material or unbound remaining reactant, increasing the specificity of the detection reaction.
(5) Application of the microporous membrane: since the pore size of the membrane is specified based on the diameter of the test substance (e.g., the pore size of a given membrane is generally 8um for CTC detection, 5um for leukemia detection, 0.8um for tubercle bacillus detection, and 0.4um nitrocellulose membrane for antigen, pathogen, or antibody detection on a protein chip, if necessary)
(6) The physical method captures and enriches various types of malignant cells (cancer cells, sarcoma cells, hematopathy cells and melanoma cells) and pathogenic organisms including bacteria, fungi, parasites, viruses and other pathogenic organisms.
(7) The detection samples are various in types, including blood, urine, saliva, cerebrospinal fluid, pleural effusion and ascites, and almost all detection samples which can be obtained by human and animals are summarized.
(8) The method is simple and rapid, and the cytopathology or etiology detection result can be obtained within 2 hours.
The utility model discloses based on malignant cell and the pathogenic organism in the detection body fluid, invent nucleated cell enrichment and dyeing integrated device and be used for the malignant cell of body fluid (especially blood) and the capture enrichment and the dyeing appraisal of pathogenic organism. The purposes of early detection and identification of malignant cells appearing in body fluid, guidance of clinical medication, and detection of metastasis and recurrence are achieved. The method achieves the purposes of early and accurate detection and identification of pathogenic organisms, emergency inspection of infectious diseases, immediate inspection of ICU patients, timely screening of major public infectious diseases and the like.
Drawings
FIG. 1 is a schematic structural view of the reaction apparatus integrating cell blotting, enrichment, and blotting and dyeing provided by the present invention.
Fig. 2 is a schematic diagram of a microporous membrane structure of the cell blotting enrichment and blotting and dyeing integrated reaction device provided by the present invention.
Wherein: 1 is microporous membrane, 2 is fixed piece, 3 is liftable objective table, 4 is base, 5 is storage bin, 6 is reaction tank, 7 is upper cover, 8 is socket, 10 is cell, 11 is the infiltration hole.
Detailed Description
The present invention is further described with reference to the following figures and specific examples, it should be understood by those skilled in the art that the following descriptions are not intended to limit the present invention in any way.
Referring to fig. 1, a cell blotting enrichment and blotting dyeing integrated reaction device comprises a microporous membrane 1, wherein the microporous membrane 1 is provided with a fixing piece 2, and the fixing piece 2 is provided with a reaction tank 6; an upper cover 7 is arranged on the reaction tank 6; a liftable objective table 3 is arranged on one side of the fixed part 2, and a storage bin (water absorption device) 5 is arranged below the fixed part 2; the storage bin 5 is filled with a water-absorbing substance. The liftable objective table 3 is connected with the base 4; a storage bin 5 is provided in the base 4.
In a specific case, the microporous membrane 1, the reaction tank 6 and the storage bin 5 are coaxially arranged up and down (or aligned). The inner diameter of the reaction tank 6 is equal to the diameter of the microporous membrane 1. The outer diameter of the reaction tank 6 is equal to the inner diameter of the upper lid 7. The ratio of the height of the reaction tank 6 to the height of the upper cover 7 is 1: (1.1-1.2). The ratio of the inner diameter of the storage bin 5 to the inner diameter of the reaction tank 6 is (1.5-1.6): 1.
referring specifically to fig. 2, the microporous membrane 1 may be specifically a polycarbonate membrane, a polyethersulfone membrane, a nitrocellulose membrane, or the like, and the microporous membrane 1 has a permeation pore 11, typically having a pore size of 3-8 microns, depending on the size of the enriched nucleated cells 10 to be retained.
The microporous membrane 1 is connected with a fixing member 2. The fixing member 2 can be made in two ways:
(1) ordinary glass formula: making holes on two glass sheets, adhering or clamping the microporous membrane 1 in the middle of the holes, and supporting and clamping the microporous membrane 1 by using an annular pad; one of the slides is bonded with a reaction tank 6 to be filled with liquid for adding body fluid or reaction liquid.
(2)3D prints the glass formula: the device is divided into an upper piece and a lower piece with proper apertures, the upper piece is directly printed with a reaction tank 6 containing liquid above the apertures, the microporous membrane 1 is adhered or clamped between the two apertures of the upper piece and the lower piece, and the microporous membrane 1 is supported and clamped by a ring-shaped pad.
The fixing part 2 of the common glass sheet type or the 3D printing glass sheet type can be made of high polymer materials, metal materials, ceramics or composite materials and the like.
The fixing piece 2 is connected with the liftable object stage 3 through the socket 8. The lifting object stage 3 can be lifted in an electric control mode, a pneumatic control mode or a hydraulic control mode.
The water-absorbing material can be absorbent paper (including filter paper, sanitary napkin, toilet paper, etc.), absorbent sponge, absorbent resin or absorbent cotton.
The following describes a method for integrating cell enrichment and blotting and dyeing by using the reaction apparatus of the present invention by using specific embodiments.
Example 1: peripheral whole blood CTC blotting enrichment and blotting immunofluorescence staining
From current literature and reagent work, CTCs include malignant cells (and possibly hematological disease cells) such as cancer cells, sarcoma cells, and melanoma cells in circulating blood. Therefore, capture enrichment and staining have important clinical significance for identifying CTCs in blood.
(1) The fixed microporous membrane device is selected according to the size of the experimental cell, and a fixed device of a polycarbonate membrane (which can be 8 microns or 3 microns) with the thickness of 5 microns is generally selected and is arranged on the water absorption device filled with the water absorption material.
(2) Adding a proper amount of microporous membrane activating solution on the microporous membrane for suction filtration.
(3) And adding a proper amount of whole blood suspended by the cell suspension on the microporous membrane, and carrying out suction printing to enrich and retain nucleated cells.
(4) And adding a proper amount of erythrocyte lysate to the microporous membrane to lyse a few erythrocytes on the microporous membrane.
(5) And adding a proper amount of cell suspension on the microporous membrane, and washing the microporous membrane.
(6) Adding methanol to fix the target cells, and separating from the water absorption device. After 5-10 minutes, the cover is covered on a water absorption device to absorb and filter liquid.
(7) And (4) separating from the water absorption device. Adding 0.1-0.2% Triton X100 and 1-10% BSA (or calf serum), and permeabilizing and blocking the cells and microporous membrane. The time is 10-30 minutes, and the cover is covered on a water absorption device to absorb and filter liquid.
(8) And (4) separating from the water absorption device. Adding monoclonal fluorescent labeled antibody, dyeing for 30-60 min, and covering the water absorbing device to suck and filter the liquid.
(9) And (4) separating from the water absorption device. Adding DAPI for counterdyeing for 5-10 min, and covering with a water absorption device to suck and filter liquid. Then, the mixture was washed with PBS (pH7.20.01mol/L) 1 to 3 times and filtered by suction.
(10) And (4) observing the result by a fluorescence microscope or a confocal microscope.
(11) The CTC trapped by enrichment and interception can be further subjected to gene detection and analysis.
Example 2: CTC blotting enrichment and blotting immunofluorescence staining after peripheral blood lysis erythrocyte
This example is different from example 1 in that blood is treated with a red blood cell lysis solution to lyse red blood cells, and then (1) the liquid after lysing red blood cells is subjected to blotting enrichment and blotting staining according to the blotting staining procedure of example 1; or (2) centrifuging the liquid after lysing the red blood cells at 12000 rpm for 5 minutes, and after completely sucking the liquid, adding the cell suspension to perform blotting enrichment and blotting staining according to example 1.
Example 3: peripheral blood CTC (cell-specific CTC) suction enrichment and blotting chemical staining
(1) The fixed microporous membrane device is selected according to the size of the experimental cell, and a fixed device of a polycarbonate membrane (which can be 8 microns or 3 microns) with the thickness of 5 microns is generally selected and is arranged on the water absorption device filled with the water absorption material.
(2) Adding a proper amount of microporous membrane activating solution on the microporous membrane for suction filtration.
(3) And adding a proper amount of whole blood suspended by the cell suspension on the microporous membrane, and carrying out suction printing to enrich and retain nucleated cells.
(4) And adding a proper amount of erythrocyte lysate to the microporous membrane to lyse a few erythrocytes on the microporous membrane.
(5) And adding a proper amount of cell suspension on the microporous membrane, and washing the microporous membrane.
(6) The methanol fixes the target cells and leaves the water absorption device. After 5-10 minutes, the cover is covered on a water absorption device to absorb and filter liquid.
(7) And (4) separating from the water absorption device. Adding a Ryje's staining solution for staining for 1-3 minutes, and adding a PBS buffer solution for staining for 5-20 minutes. The staining solution was aspirated and washed 1-3 times with PBS.
(8) And observing the result by using a common optical microscope.
(9) If necessary, treating blood with erythrocyte lysate to lyse erythrocytes, and then (1) subjecting the liquid with lysed erythrocytes to blotting enrichment and blotting staining according to the blotting staining procedure of example 1; or (2) the liquid after lysing the erythrocytes was centrifuged at 12000 rpm for 5 minutes, and after the liquid was aspirated, the cell suspension was added, and chemical staining was performed according to this example.
(10) The CTC trapped by enrichment and interception can be further subjected to gene detection and analysis.
Example 4: breast and ascites malignant tumor cell blotting enrichment and blotting immunofluorescence staining
(1) The fixed microporous membrane device is selected according to the size of the experimental cell, and a fixed device of a polycarbonate membrane (which can be 8 microns or 3 microns) with the thickness of 5 microns is generally selected and is arranged on the water absorption device filled with the water absorption material.
(2) Adding a proper amount of microporous membrane activating solution on the microporous membrane for suction filtration.
(3) Adding appropriate amount of hydrothorax and ascites suspended by cell suspension on microporous membrane, sucking and printing, and enriching and intercepting nucleated cells.
Note that: if the chest and abdominal water volume is large, firstly, centrifuging the chest and abdominal water to precipitate cells, taking the precipitate, adding a proper amount of cell suspension to suspend the cells, and then adding a microporous membrane to absorb and print the enriched nucleated cells.
(4) And adding a proper amount of erythrocyte lysate to the microporous membrane to lyse a few erythrocytes on the microporous membrane.
(5) And adding a proper amount of cell suspension on the microporous membrane, and washing the microporous membrane.
(6) Adding methanol to fix the target cells, and separating from the water absorption device. After 5-10 minutes, the cover is covered on a water absorption device to absorb and filter liquid.
(7) And (4) separating from the water absorption device. Adding 0.1-0.2% Triton X100 and 1-10% BSA (or calf serum), and permeabilizing and blocking the cells and microporous membrane. The time is 10-30 minutes, and the cover is covered on a water absorption device to absorb and filter liquid.
(8) And (4) separating from the water absorption device. Adding monoclonal fluorescent labeled antibody, dyeing for 30-60 min, and covering the water absorbing device to suck and filter the liquid.
(9) And (4) separating from the water absorption device. Adding DAPI for counterdyeing for 5-10 min, and covering with a water absorption device to suck and filter liquid. Then, the mixture was washed with PBS (pH7.20.01mol/L) 1 to 3 times and filtered by suction.
(10) And (4) observing the result by a fluorescence microscope or a confocal microscope.
(11) The CTC trapped by enrichment and interception can be further subjected to gene detection and analysis.
Example 5: capture enrichment and staining identification of cervical swab or cervical scraping cancer cells
The cervical cancer is a common malignant tumor in gynecology, cervical cancer cells are detected by using a cervical swab or a cervical scraping strip, the cervical cancer is screened and diagnosed at an early stage, medication is guided, and the cervical cancer diagnostic kit has an important clinical application value.
(1) A 5 micron polycarbonate membrane mounting device is typically selected and mounted on a water absorbing device containing a water absorbing material.
(2) Adding a proper amount of microporous membrane activating solution on the microporous membrane for suction filtration.
(3) And adding a proper amount of cell suspension to suspend the cell liquid of the cervical swab or the cervical scraping blade on the microporous membrane, and sucking, printing, enriching and intercepting the nucleated cells.
(4) And adding a proper amount of erythrocyte lysate to the microporous membrane to lyse a few erythrocytes on the microporous membrane.
(5) And adding a proper amount of cell suspension on the microporous membrane, and washing the microporous membrane.
(6) Adding methanol to fix the target cells, and separating from the water absorption device. After 5-10 minutes, the cover is covered on a water absorption device to absorb and filter liquid.
(7) And (4) separating from the water absorption device. Adding 0.1-0.2% Triton X100 and 1-10% BSA (or calf serum), and permeabilizing and blocking the cells and microporous membrane. The time is 10-30 minutes, and the cover is covered on a water absorption device to absorb and filter liquid.
(8) And (4) separating from the water absorption device. Adding monoclonal fluorescent labeled antibody, dyeing for 30-60 min, and covering the water absorbing device to suck and filter the liquid.
(9) And (4) separating from the water absorption device. Adding DAPI for counterdyeing for 5-10 min, and covering with a water absorption device to suck and filter liquid. Then, the mixture was washed with PBS (pH7.20.01mol/L) 1 to 3 times and filtered by suction.
(10) And (4) observing the result by a fluorescence microscope or a confocal microscope.
(11) The CTC trapped by enrichment and interception can be further subjected to gene detection and analysis.
Example 6: detection of mycobacterium tuberculosis in body fluids (blood, cerebrospinal fluid, pleural effusion, etc.)
(1) Collecting 2-5 ml of blood by using an EDTA anticoagulant tube, and absorbing plasma after red blood cells of the blood naturally settle; or anticoagulation 1000-.
(2) Adding appropriate amount of cell suspension to suspend blood cell volume to original volume, adding anhydrous alcohol to 70% concentration, and killing Bacillus tuberculosis in cells at room temperature for 30 min. Then centrifuged and the cell suspension added to the original volume.
(3) Adding 2-10% sodium hypochlorite solution into blood plasma, hydrothorax, cerebrospinal fluid, etc. at a ratio of 2-6:1 to blood plasma, cerebrospinal fluid and hydrothorax, and sterilizing tubercle bacillus at room temperature for 30 min. 4000-10000 r/g/separating the core for a certain time to precipitate the tubercle bacillus.
(4) Suspending 1 ml of the suspended blood cells or pleural fluid and cerebrospinal fluid with a proper amount of cell suspension.
(5) Plasma, pleural fluid, and cerebrospinal fluid with 0.8 μm microporous membrane; blood cells were treated with a 5 micron microporous membrane. And adding a proper amount of cell suspension on the microporous membrane, and washing the microporous membrane.
(6) Adding methanol to fix the target cells, and separating from the water absorption device. After 5-10 minutes, the cover is covered on a water absorption device to absorb and filter liquid.
(7) And (4) separating from the water absorption device. Adding 0.1-0.2% Triton X100 and 1-10% BSA (or calf serum), and permeabilizing and blocking the cells and microporous membrane. The time is 10-30 minutes, and the cover is covered on a water absorption device to absorb and filter liquid.
(8) And (4) separating from the water absorption device. Adding monoclonal fluorescence labeling antibody, and dyeing for 30-60 minutes; or adding gold amine O fluorescent dye to dye for 10-20 min. The pressure cover is covered on the water absorption device to absorb and filter liquid.
(9) And (4) separating from the water absorption device. Adding DAPI for counterdyeing for 5-10 min, and covering with a water absorption device to suck and filter liquid. Then, the mixture was washed with PBS (pH7.20.01mol/L) 1 to 3 times and filtered by suction.
(10) And (4) observing the result by a fluorescence microscope or a confocal microscope.
(11) The trapped tubercle bacillus can be further subjected to gene detection or gene drug sensitivity test; the trapped tubercle bacillus can also be acid-fast stained.
(12) The morphology and the fluorescence color of the tubercle bacillus are observed by a fluorescence microscope.
The utility model discloses a beneficial effect and innovation point do:
(1) uptake of trapped material based on water-absorbing material: when the cell suspension passes through the microporous membrane, cells with the pore size larger than that of the membrane are intercepted in the reaction container and adsorbed on the membrane surface due to the adoption of an attraction filtration interception mode. The water-absorbing material has stable absorption speed, soft absorption force, slow speed and no pressure, so that the cells are attached to the surface of the membrane.
(2) Low-speed filtration and interception: the shearing force for intercepting the cells by low-speed filtration is small, and the morphological structure of the cells is not damaged.
(3) The blotting reaction time can be set: the reactant (antigen antibody, marker, etc.) is contacted with the cell sufficiently and reacted for a certain time, so as to achieve the purpose of fully reacting and fully increasing the detection sensitivity.
(4) The washing time and the number of washing times can be set: sufficient washing can remove non-specific material or unbound remaining reactant, increasing the specificity of the detection reaction.
(5) Application of the microporous membrane: since the pore size of the membrane is specified based on the diameter of the test substance, for example, the pore size of a given membrane is generally 8um for detecting CTC, 5um for detecting leukemia cells, 0.8um for detecting tubercle bacillus, and the pore size of a microporous membrane for retaining other microbial cells is used as required, and a nitrocellulose membrane of 0.4um for detecting antigen, pathogen or antibody is used for the protein chip.
(6) The physical method captures and enriches various types of malignant cells (cancer cells, sarcoma cells, hematopathy cells and melanoma cells) and pathogenic organisms including bacteria, fungi, parasites, viruses and other pathogenic organisms.
(7) The detection samples are various in types, including blood, urine, saliva, cerebrospinal fluid, pleural effusion and ascites, and almost all detection samples which can be obtained by human and animals are summarized.
(8) The method is simple and rapid, and the cytopathology or etiology detection result can be obtained within 2 hours.
The utility model discloses based on malignant cell and the pathogenic organism in the detection body fluid, invent nucleated cell enrichment and dyeing integrated device and be used for the malignant cell of body fluid (especially blood) and the capture enrichment and the dyeing appraisal of pathogenic organism. The purposes of early detection and identification of malignant cells appearing in body fluid, guidance of clinical medication, and detection of metastasis and recurrence are achieved. The method achieves the purposes of early and accurate detection and identification of pathogenic organisms, emergency inspection of infectious diseases, immediate inspection of ICU patients, timely screening of major public infectious diseases and the like.
Claims (10)
1. The utility model provides a cell inhales seal enrichment and inhales seal dyeing integration reaction unit which characterized in that: comprises a microporous membrane (1), wherein the microporous membrane (1) is provided with a fixing piece (2), and the fixing piece (2) is provided with a reaction tank (6); an upper cover (7) is arranged on the reaction tank (6); a liftable objective table (3) is arranged on one side of the fixing piece (2), and a storage bin (5) is arranged below the fixing piece (2); the storage bin (5) is filled with water-absorbing substances.
2. The integrated reaction device for cell blotting enrichment and blotting dyeing as claimed in claim 1, wherein the microporous membrane (1) comprises polycarbonate membrane, polyethersulfone membrane, and nitrocellulose membrane.
3. The integrated reaction device for cell blotting enrichment and blotting as claimed in claim 1, wherein: the microporous membrane (1) is connected with the fixing piece (2); the fixing piece (2) is of a common glass sheet type: making holes on two glass sheets, adhering or clamping the microporous membrane (1) in the middle of the holes, and supporting and clamping the microporous membrane by using an annular pad; one of the slides is bonded with a reaction tank (6) to be filled with liquid for adding body fluid or reaction liquid.
4. The integrated reaction device for cell blotting enrichment and blotting as claimed in claim 1, wherein: the microporous membrane (1) is connected with the fixing piece (2); fixing piece (2) for 3D print the glass formula: the device is divided into an upper piece and a lower piece with proper apertures, the upper piece is directly printed with a reaction tank (6) to be filled with liquid above the apertures, a microporous membrane (1) is adhered or clamped between the two apertures of the upper piece and the lower piece, and an annular pad is used for supporting and clamping the microporous membrane.
5. The integrated reaction device for cell blotting enrichment and blotting as claimed in claim 1, wherein: the inner diameter of the reaction tank (6) is equal to the diameter of the microporous membrane (1).
6. The integrated reaction device for cell blotting enrichment and blotting as claimed in claim 1, wherein: the outer diameter of the reaction tank (6) is equal to the inner diameter of the upper cover (7).
7. The integrated reaction device for cell blotting enrichment and blotting as claimed in claim 1, wherein: the ratio of the height of the reaction tank (6) to the height of the upper cover (7) is 1: (1.1-1.2).
8. The integrated reaction device for cell blotting enrichment and blotting as claimed in claim 1, wherein: the ratio of the inner diameter of the storage bin (5) to the inner diameter of the reaction tank (6) is (1.5-1.6): 1.
9. the integrated reaction device for cell blotting enrichment and blotting as claimed in claim 1, wherein: the fixing piece (2) is connected with the liftable object stage (3) through the socket (8).
10. The integrated reaction device for cell blotting enrichment and blotting as claimed in claim 1, wherein: the lifting object stage (3) is connected with the base (4); the storage bin (5) is arranged in the base (4).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201920564841.3U CN209940977U (en) | 2019-04-24 | 2019-04-24 | Cell suction printing enrichment and suction printing dyeing integrated reaction device |
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| CN201920564841.3U CN209940977U (en) | 2019-04-24 | 2019-04-24 | Cell suction printing enrichment and suction printing dyeing integrated reaction device |
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| CN209940977U true CN209940977U (en) | 2020-01-14 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111855333A (en) * | 2019-04-24 | 2020-10-30 | 青岛北大新世纪言鼎生物医学科技有限公司 | Integrated reaction device and method for absorbing, printing, enriching and absorbing, printing and dyeing nucleated cells in body fluid |
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2019
- 2019-04-24 CN CN201920564841.3U patent/CN209940977U/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111855333A (en) * | 2019-04-24 | 2020-10-30 | 青岛北大新世纪言鼎生物医学科技有限公司 | Integrated reaction device and method for absorbing, printing, enriching and absorbing, printing and dyeing nucleated cells in body fluid |
| CN111855333B (en) * | 2019-04-24 | 2023-10-03 | 青岛言鼎生物医疗科技有限公司 | Device and method for integrating enrichment and dye-printing absorption and dyeing of nucleated cells in body fluid |
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