CN113174361B - Pathological cell separation adsorption type extraction method - Google Patents
Pathological cell separation adsorption type extraction method Download PDFInfo
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Abstract
The invention provides a pathological cell separation adsorption type extraction method, which belongs to the field of cell extraction, and comprises the following steps: s1, collecting pleurisy chest water and storing and transporting; s2, preparing a mixed solution; s3, mounting and preparing an adsorption type extracting device and an adsorption layer; s4, adding the mixed solution prepared in the step S2, and carrying out separation adsorption type extraction through an adsorption layer; s5, disassembling the adsorption layer, and extracting the separated pathological cells; s6, transferring the extracted pathological cells to a culture medium for purification culture, and freezing and storing. The invention maintains the activity of cells to the maximum extent by the structural design of the shell with the adsorption layer, does not destroy the structure of the cells, and can separate the non-relevant cells such as the white blood cells, the red blood cells and the blood platelets respectively in a layered manner by a multi-layer adsorption mode, thereby being convenient for purifying pathological cells, having market prospect and being suitable for popularization.
Description
Technical Field
The invention relates to the field of cell extraction, in particular to a pathological cell separation adsorption type extraction method.
Background
Pleurisy can be caused by infections (bacteria, viruses, mold, amoeba, lung flukes, etc.) and infectious agents, such as tumors, allergies, chemical and traumatic diseases. Among pleurisy caused by bacterial infection, tuberculous pleurisy is most common. Common diseases causing pleurisy: pulmonary infarction, cancer, tuberculosis, rheumatoid arthritis, systemic lupus erythematosus, parasitic infections (such as amoebae), pancreatitis, injuries (such as rib fractures), irritants from airways or other sites to the pleura (such as asbestos), drug allergies (such as hydralazine, procainamide, isoniazid, phenytoin, chlorpromazine) and the like.
Pathological examination has been applied to a great deal of clinical work and scientific research. The cadaver pathological examination and the operation pathological examination are mainly carried out in clinical aspects. The purpose of pathological examination of operation is to definitely diagnose and verify the preoperative diagnosis, and improve the clinical diagnosis level; and secondly, after the diagnosis is clear, the next treatment scheme and the estimated prognosis can be determined, so that the clinical treatment level is improved, and a large amount of valuable scientific research data can be obtained through clinical pathological analysis.
Conventional pleurisy chest water pathological cell extraction needs to be subjected to centrifugation, precipitation, fixation and slicing, and a high-speed centrifugation mode is often adopted when the pathological cells are separated, but the mode inevitably causes a certain degree of damage to the activity of some cells. Therefore, we propose a pathological cell separation adsorption extraction method to solve the above problems.
Disclosure of Invention
1. Technical problem to be solved
Aiming at the problems existing in the prior art, the invention aims to provide a pathological cell separation adsorption type extraction method, which can separate relevant pathological extracting solutions in an adsorption type under normal pressure and low temperature through a shell structure design with an adsorption layer in a separation adsorption mode, so that the activity of cells is kept to the maximum extent, the structure is not destroyed, and the method can separate non-relevant cells such as leucocytes, erythrocytes and blood platelets in a multi-layer adsorption mode respectively, thereby being convenient for purifying the pathological cells, having market prospect and being suitable for popularization.
2. Technical proposal
In order to solve the problems, the invention adopts the following technical scheme.
A pathological cell separation adsorption type extraction method comprises the following steps:
s1, collecting pleurisy chest water in an aseptic environment of an operating room, storing the chest water in an aseptic storage box, labeling and bar-coding, keeping the temperature of the storage box at 4-10 ℃, and delivering the chest water to a laboratory within 24 hours;
s2, pleurisy chest water and normal saline in the storage and transportation box are mixed according to the proportion of 3:1, uniformly mixing the materials in proportion to obtain a mixed solution;
s3, disassembling the adsorption type extraction device, placing the adsorption type extraction device under ultraviolet sterilization light, sterilizing for 30min, and installing the adsorption layer in the adsorption type extraction device after sterilization is completed;
s4, pressing the shell of the adsorption type extraction device through external force to enable the interior of the shell to be in a negative pressure state, adding the mixed liquid prepared in the step S2 along a liquid inlet of the adsorption type extraction device, and separating and adsorbing the mixed liquid through an adsorption layer;
s5, disassembling the adsorption layers, and extracting the separated pleurisy pathological cells from the corresponding adsorption layers according to the diameters of the pathological cells;
s6, transferring the extracted pathological cells to a culture medium for purification culture, and freezing and storing.
The invention separates the related pathological extracting solution in an adsorption mode under normal pressure and low temperature by virtue of the shell structure design with an adsorption layer, so that the activity of cells is maintained to the maximum extent, the structure is not destroyed, and the method can separate the non-related cells such as leucocytes, erythrocytes and blood platelets in a layered manner by virtue of a multi-layer adsorption mode, thereby being convenient for purifying the pathological cells, having market prospect and being suitable for popularization.
Further, the top of adsorption type extraction element is connected with the cap through the helicitic texture is detachable, the casing is round platform type structure, the adsorbed layer sets up between cap and casing, the liquid inlet sets up on the cap, the base has been cup jointed in the casing bottom slip, the damping spring has been cup jointed between base and the casing. Through pressing the air compression that makes between casing and the base of casing, when throwing into the mixed liquid, under damping spring's storage's elastic potential energy effect, the casing gradually resets, makes the adsorbed layer one side atmospheric pressure that is equipped with the mixed liquid be greater than its opposite side atmospheric pressure, and then promotes the passing efficiency that the mixed liquid passed the adsorbed layer, has effectively promoted extraction rate.
Further, the casing internal fixation has presses the support, it is tubular structure to press the support, it is equipped with the straw to press the top grafting of support, the straw includes rubber head and straw mouth, still be fixed with on the straw with press support assorted spacing ring. Through the structural design of the straw with the rubber head, the straw mouth and the limiting ring, the straw is convenient to install, meanwhile, the separated pathological cells are also convenient to collect at any time and rapidly, when the diameter of the pathological cells to be extracted is positioned between the adjacent adsorption structure apertures, the corresponding adsorption layers are only required to be disassembled, the pathological cells are extracted by utilizing the straw, the use convenience is improved, and the working efficiency is effectively improved.
Further, the bottom of the shell is provided with a liquid passing groove, the liquid collecting groove is fixed at the inner bottom of the base, a liquid guiding groove for guiding extracting liquid into the liquid collecting groove is fixed on the inner wall of the base, and a sealing gasket matched with the liquid passing groove is arranged at the top of the liquid guiding groove. Through the structural design of the liquid guide groove with the sealing gasket and the liquid accumulation groove, when the diameter of pathological cells to be extracted is smaller than the minimum adsorption aperture of the adsorption layer, the pathological cells flow into the liquid accumulation groove through the guide of the liquid guide groove, after collection is finished, the rubber head of the suction pipe can be directly pressed, and the pathological cells are collected from the liquid accumulation groove to be transferred, so that the working efficiency is effectively improved.
Further, the adsorption layer comprises an upper connecting ring, a lower connecting ring and an adsorption core, wherein the adsorption core is divided into a plurality of layers of adsorption structures with different apertures, the layers of adsorption structures are sequentially sleeved and arranged in a conical shape, and the conical surface of the adsorption structure is arranged in a forty-five gradient manner.
Further, the conical inner wall equidistance of adsorption structure is equallyd divide fixedly to be equipped with a plurality of baffles, the baffle sets up perpendicularly with adsorption structure's conical inner wall. Through the adsorption structure that forty-five slope set up and rather than vertically baffle structural design, promote adsorption structure's structural strength on the one hand, on the other hand, when throwing into the mixed liquid, through the blocking of baffle, make the mixed liquid divide into a plurality of annular group, effectively promoted adsorption structure's effective adsorption area, the design of baffle also is favorable to extracting the pathological cell that separates down simultaneously.
Further, one side of the upper connecting ring, which is far away from the lower connecting ring, is provided with an annular buckle, and the shell cover are provided with buckle grooves matched with the annular buckle. Through buckle groove and annular buckle, accessible cap and the involution of casing, install fast and dismantle the adsorbed layer, promoted the convenience of using.
Further, clamping and connecting tensioning device have between last go up the go-between and adsorb the core, tensioning device includes spring leaf and tensioning rubber, the spring leaf is the ring structure of section V style of calligraphy, tensioning rubber is fixed between the V type opening of spring leaf, the top side of spring leaf is fixed with the bottom side glue joint of last go-between, the upper end and the lower extreme of adsorb the core are fixed with spring leaf bottom, lower go-between top glue joint respectively. Through the structural design of the tensioning device with tensioning rubber and a spring piece, the tensioning rubber is utilized to tighten the spring piece, the upper end and the lower end of the adsorption core can be effectively tensioned, the structural stability of the adsorption core is ensured, the collapse deformation of the adsorption core is avoided, and the adsorption efficiency is influenced.
Further, the multi-layer adsorption structure of the adsorption core sequentially comprises a leukocyte adsorption layer, a erythrocyte adsorption layer and a platelet adsorption layer from inside to outside, wherein the leukocyte adsorption layer is of a nylon fiber structure, the adsorption aperture of the leukocyte adsorption layer is 10-20 mu m, the erythrocyte adsorption layer is of a neutral punching adsorption gum structure, the adsorption aperture of the erythrocyte adsorption layer is 7-8 mu m, the platelet adsorption layer is of a single-wall carbon nano tube non-woven membrane structure, and the adsorption aperture of the platelet adsorption layer is 2-3 mu m.
Further, temperature detecting device and constant temperature equipment are arranged at the top of the shell cover, and the detecting end of the temperature detecting device and the output end of the constant temperature equipment extend into the shell. Through the design of the temperature detection device and the shell cover of the constant temperature device, the low temperature state in the shell can be effectively maintained, and the activity of cells can be maintained to the maximum extent.
3. Advantageous effects
Compared with the prior art, the invention has the advantages that:
(1) According to the scheme, through the shell structure design with the adsorption layer, the related pathological extracting solution is subjected to adsorption separation in a separation adsorption mode under normal pressure and low temperature, the activity of cells is maintained to the maximum extent, the structure is not damaged, and the method can also be used for respectively carrying out layering separation on the non-related cells such as the white blood cells, the red blood cells and the blood platelets in a multi-layer adsorption mode, so that the purification of the pathological cells is facilitated, and the method has market prospects and is suitable for popularization.
(2) Through pressing the air compression that makes between casing and the base of casing, when throwing into the mixed liquid, under damping spring's storage's elastic potential energy effect, the casing gradually resets, makes the adsorbed layer one side atmospheric pressure that is equipped with the mixed liquid be greater than its opposite side atmospheric pressure, and then promotes the passing efficiency that the mixed liquid passed the adsorbed layer, has effectively promoted extraction rate.
(3) Through the structural design of the straw with the rubber head, the straw mouth and the limiting ring, the straw is convenient to install, meanwhile, the separated pathological cells are also convenient to collect at any time and rapidly, when the diameter of the pathological cells to be extracted is positioned between the adjacent adsorption structure apertures, the corresponding adsorption layers are only required to be disassembled, the pathological cells are extracted by utilizing the straw, the use convenience is improved, and the working efficiency is effectively improved.
(4) Through the structural design of the liquid guide groove with the sealing gasket and the liquid accumulation groove, when the diameter of pathological cells to be extracted is smaller than the minimum adsorption aperture of the adsorption layer, the pathological cells flow into the liquid accumulation groove through the guide of the liquid guide groove, after collection is finished, the rubber head of the suction pipe can be directly pressed, and the pathological cells are collected from the liquid accumulation groove to be transferred, so that the working efficiency is effectively improved.
(5) Through the adsorption structure that forty-five slope set up and rather than vertically baffle structural design, promote adsorption structure's structural strength on the one hand, on the other hand, when throwing into the mixed liquid, through the blocking of baffle, make the mixed liquid divide into a plurality of annular group, effectively promoted adsorption structure's effective adsorption area, the design of baffle also is favorable to extracting the pathological cell that separates down simultaneously.
(6) Through buckle groove and annular buckle, accessible cap and the involution of casing, install fast and dismantle the adsorbed layer, promoted the convenience of using.
(7) Through the structural design of the tensioning device with tensioning rubber and a spring piece, the tensioning rubber is utilized to tighten the spring piece, the upper end and the lower end of the adsorption core can be effectively tensioned, the structural stability of the adsorption core is ensured, the collapse deformation of the adsorption core is avoided, and the adsorption efficiency is influenced.
(8) Through the design of the temperature detection device and the shell cover of the constant temperature device, the low temperature state in the shell can be effectively maintained, and the activity of cells can be maintained to the maximum extent.
Drawings
FIG. 1 is a flow chart of the method of the present invention;
FIG. 2 is a schematic diagram of an adsorption type extraction apparatus according to the present invention;
FIG. 3 is a schematic cross-sectional view of an adsorption type extraction apparatus according to the present invention;
FIG. 4 is an enlarged schematic view of the portion A in FIG. 3;
FIG. 5 is a schematic diagram of an explosion structure of an adsorption type extraction device according to the present invention;
FIG. 6 is a schematic diagram of an adsorption layer according to the present invention;
FIG. 7 is a schematic diagram of an exploded structure of an adsorption layer according to the present invention;
FIG. 8 is a schematic cross-sectional view of a tensioner of the present invention;
FIG. 9 is a schematic view of a suction tube according to the present invention;
fig. 10 is a schematic structural view of a cover according to the present invention.
The reference numerals in the figures illustrate:
the device comprises a shell 1, a base 11, a liquid accumulation groove 12, a pressing support 13, a liquid passing groove 14, a liquid guide groove 15, a sealing gasket 151, a damping spring 16, a shell cover 2, a liquid inlet 21, a temperature detection device 22, a constant temperature device 23, a suction pipe 3, a suction pipe nozzle 31, a rubber head 32, a limiting ring 33, an adsorption layer 4, an upper connecting ring 41, a lower connecting ring 42, an adsorption core 43, a baffle 44, a tensioning device 45, a spring piece 451, tensioning rubber 452 and an annular buckle 46.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention; it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments, and that all other embodiments obtained by persons of ordinary skill in the art without making creative efforts based on the embodiments in the present invention are within the protection scope of the present invention.
In the description of the present invention, it should be noted that the positional or positional relationship indicated by the terms such as "upper", "lower", "inner", "outer", "top/bottom", etc. are based on the positional or positional relationship shown in the drawings, are merely for convenience of describing the present invention and simplifying the description, and do not indicate or imply that the apparatus or elements 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 the like, 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 should be noted that, unless explicitly specified and limited otherwise, the terms "mounted," "configured to," "engaged with," "connected to," and the like are to be construed broadly, and may be either fixedly connected, detachably connected, or integrally connected, for example; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements. The specific meaning of the above terms in the present invention will be understood in specific cases by those of ordinary skill in the art.
Example 1:
referring to fig. 1-4, a pathological cell separation adsorption extraction method comprises the following steps:
s1, collecting pleurisy chest water in an aseptic environment of an operating room, storing the chest water in an aseptic storage box, labeling and bar-coding, keeping the temperature of the storage box at 4-10 ℃, and delivering the chest water to a laboratory within 24 hours;
s2, pleurisy chest water and normal saline in the storage and transportation box are mixed according to the proportion of 3:1, uniformly mixing the materials in proportion to obtain a mixed solution;
s3, disassembling the adsorption type extraction device, placing under ultraviolet sterilization light, sterilizing for 30min, and installing the adsorption layer 4 in the adsorption type extraction device after sterilization is completed;
s4, pressing the shell 1 of the adsorption type extraction device by external force to enable the interior of the shell 1 to be in a negative pressure state, adding the mixed liquid prepared in the step S2 along the liquid inlet 21 of the adsorption type extraction device, and separating and adsorbing the mixed liquid through the adsorption layer 4;
s5, disassembling the adsorption layers 4, and extracting the separated pleurisy pathological cells from the corresponding adsorption layers 4 according to the diameters of the pathological cells;
s6, transferring the extracted pathological cells to a culture medium for purification culture, and freezing and storing.
The invention separates the relevant pathological extracting solution in an adsorption mode under normal pressure and low temperature by virtue of the structural design of the shell 1 with the adsorption layer 4, so that the activity of cells is maintained to the maximum extent, the structure is not destroyed, and the method can separate the non-relevant cells such as leucocytes, erythrocytes and platelets in a layered manner by virtue of a multi-layer adsorption mode, thereby being convenient for purifying the pathological cells, having market prospect and being suitable for popularization.
Referring to fig. 2-4, the top of the adsorption type extracting device is detachably connected with a casing cover 2 through a threaded structure, a casing 1 is in a truncated cone structure, an adsorption layer 4 is arranged between the casing cover 2 and the casing 1, a liquid inlet 21 is formed in the casing cover 2, a base 11 is sleeved at the bottom of the casing 1 in a sliding manner, and a damping spring 16 is clamped between the base 11 and the casing 1. Through pressing the air compression that makes between casing 1 and the base 11 of casing 1, when throwing into the mixed liquor, under the elastic potential energy effect of the storage of damping spring 16, casing 1 gradually resets, makes the adsorbed layer 4 one side atmospheric pressure that is equipped with the mixed liquor be greater than its opposite side atmospheric pressure, and then promotes the passing efficiency that the mixed liquor passed adsorbed layer 4, has effectively promoted extraction rate.
Referring to fig. 2 and 9, a pressing bracket 13 is fixed in the housing 1, the pressing bracket 13 is in a circular tube structure, a suction tube 3 is inserted into the top of the pressing bracket 13, the suction tube 3 comprises a rubber head 32 and a suction tube nozzle 31, and a limiting ring 33 matched with the pressing bracket 13 is fixed on the suction tube 3. Through the structural design of the straw 3 with the rubber head 32, the straw mouth 31 and the limiting ring 33, the straw 3 is convenient to arrange, meanwhile, the separated pathological cells are also convenient to collect quickly at any time, when the diameter of the pathological cells to be extracted is positioned between the adjacent adsorption structure apertures, the corresponding adsorption layer 4 is only required to be disassembled, the pathological cells are extracted by utilizing the straw 3, the use convenience is improved, and the work efficiency is effectively improved.
Referring to fig. 2, a liquid passing groove 14 is provided at the bottom of the housing 1, a liquid collecting groove 12 is fixed at the inner bottom of the base 11, a liquid guiding groove 15 for guiding the extracting liquid into the liquid collecting groove 12 is fixed at the inner wall of the base 11, and a sealing gasket 151 matched with the liquid passing groove 14 is provided at the top of the liquid guiding groove 15. Through the structural design of the liquid guide groove 15 with the sealing gasket 151 and the liquid accumulation groove 12, when the diameter of the pathological cells to be extracted is smaller than the minimum adsorption aperture of the adsorption layer 4, the pathological cells flow into the liquid accumulation groove 12 through the guide of the liquid guide groove 15, after the collection is finished, the rubber head 32 of the suction pipe 3 can be directly pressed, and the pathological cells are collected from the liquid accumulation groove 12 for transfer, so that the working efficiency is effectively improved.
Referring to fig. 4-7, the adsorption layer 4 includes an upper connecting ring 41, a lower connecting ring 42 and an adsorption core 43, the adsorption core 43 is divided into multiple layers of adsorption structures with different apertures, the multiple layers of adsorption structures are sequentially sleeved and arranged in a tapered shape, the tapered surfaces of the adsorption structures are arranged in forty-five inclination, a plurality of baffles 44 are uniformly distributed and fixed on the tapered inner walls of the adsorption structures at equal intervals, and the baffles 44 are vertically arranged with the tapered inner walls of the adsorption structures. Through the adsorption structure that forty-five slope set up and rather than vertically baffle 44 structural design, promote adsorption structure's structural strength on the one hand, on the other hand, when throwing into the mixed liquid, through the blocking of baffle 44, make the mixed liquid divide into a plurality of annular group, effectively promoted adsorption structure's effective adsorption area, the design of baffle 44 also is favorable to extracting the pathological cell that separates down simultaneously, go up go-between 41 and down go-between 42 looks one side that keeps away from all is equipped with annular buckle 46, all be equipped with on casing 1 and the cap 2 with annular buckle 46 assorted buckle groove. Through buckle groove and annular buckle 46, can utilize cap 2 and the involution of casing 1, install fast and dismantle adsorbed layer 4, promoted the convenience of using.
Referring to fig. 6-8, a tensioning device 45 is clamped between the upper connecting ring 41 and the adsorption core 43, the tensioning device 45 comprises a spring piece 451 and tensioning rubber 452, the spring piece 451 is of a circular ring structure with a V-shaped section, the tensioning rubber 452 is fixed between the V-shaped openings of the spring piece 451, the top side of the spring piece 451 is fixed with the bottom side of the upper connecting ring 41 in a gluing manner, and the upper end and the lower end of the adsorption core 43 are fixed with the bottom end of the spring piece 451 and the top end of the lower connecting ring 42 in a gluing manner respectively. Through the structural design of the tensioning device 45 with the tensioning rubber 452 and the spring piece 451, the tensioning rubber 452 is utilized to tighten the spring piece 451, the upper end and the lower end of the adsorption core 43 can be effectively tensioned, the structural stability of the adsorption core 43 is guaranteed, collapse deformation of the adsorption core 43 is avoided, adsorption efficiency is affected, a multi-layer adsorption structure of the adsorption core 43 sequentially comprises a leucocyte adsorption layer, a erythrocyte adsorption layer and a platelet adsorption layer from inside to outside, the leucocyte adsorption layer is of a nylon fiber structure, the adsorption aperture of the leucocyte adsorption layer is 10-20 mu m, the erythrocyte adsorption layer is of a neutral punching adsorption gum structure, the adsorption aperture of the erythrocyte adsorption layer is 7-8 mu m, the platelet adsorption layer is of a single-walled carbon nanotube non-woven membrane structure, and the adsorption aperture of the platelet adsorption layer is 2-3 mu m.
Referring to fig. 10, a temperature detecting device 22 and a thermostat 23 are disposed on top of the cover 2, and both a detecting end of the temperature detecting device 22 and an output end of the thermostat 23 extend into the housing 1. The low temperature state in the shell 1 can be effectively maintained by the design of the shell cover 2 of the temperature detection device 22 and the constant temperature device 23, and the activity of cells can be maintained to the maximum extent.
The above description is only of the preferred embodiments of the present invention; the scope of the invention is not limited in this respect. Any person skilled in the art, within the technical scope of the present disclosure, may apply to the present invention, and the technical solution and the improvement thereof are all covered by the protection scope of the present invention.
Claims (4)
1. A pathological cell separation adsorption type extraction method, which is characterized by comprising the following steps:
s1, collecting pleurisy chest water in an aseptic environment of an operating room, storing the chest water in an aseptic storage box, labeling and bar-coding, keeping the temperature of the storage box at 4-10 ℃, and delivering the chest water to a laboratory within 24 hours;
s2, pleurisy chest water and normal saline in the storage and transportation box are mixed according to the proportion of 3:1, uniformly mixing the materials in proportion to obtain a mixed solution;
s3, disassembling the adsorption type extraction device, placing the adsorption type extraction device under ultraviolet sterilization light, sterilizing for 30min, and installing an adsorption layer (4) in the adsorption type extraction device after sterilization is completed;
s4, pressing the shell (1) of the adsorption type extraction device by external force to enable the interior of the shell (1) to be in a negative pressure state, adding the mixed liquid prepared in the step S2 along the liquid inlet (21) of the adsorption type extraction device, and separating and adsorbing the mixed liquid through the adsorption layer (4);
s5, detaching the adsorption layers (4), and extracting the separated pleurisy pathological cells from the corresponding adsorption layers (4) according to the diameters of the pathological cells;
s6, transferring the extracted pathological cells to a culture medium for purification culture, and freezing and storing;
the top of the adsorption type extraction device is detachably connected with a shell cover (2) through a thread structure, the shell (1) is of a round table structure, the adsorption layer (4) is arranged between the shell cover (2) and the shell (1), the liquid inlet (21) is formed in the shell cover (2), the bottom of the shell (1) is in sliding sleeve connection with a base (11), a damping spring (16) is clamped between the base (11) and the shell (1), the shell (1) is pressed to enable air between the shell (1) and the base (11) to be compressed, and when mixed liquid is put into, the shell (1) can be gradually reset under the action of elastic potential energy stored by the damping spring (16) and the air pressure on one side of the adsorption layer (4) is larger than that on the other side. The novel plastic suction tube is characterized in that a pressing support (13) is fixed in the shell (1), the pressing support (13) is of a circular tube structure, a suction tube (3) is inserted into the top of the pressing support (13), the suction tube (3) comprises a rubber head (32) and a suction tube nozzle (31), and a limiting ring (33) matched with the pressing support (13) is fixed on the suction tube (3); the bottom of the shell (1) is provided with a liquid passing groove (14), a liquid accumulation groove (12) is fixed at the inner bottom of the base (11), a liquid guide groove (15) for guiding extracting liquid into the liquid accumulation groove (12) is fixed at the inner wall of the base (11), a sealing gasket (151) matched with the liquid passing groove (14) is arranged at the top of the liquid guide groove (15), and when the diameter of pathological cells to be extracted is smaller than the minimum adsorption aperture of the adsorption layer (4), the pathological cells can flow into the liquid accumulation groove (12) under the guiding action of the liquid guide groove (15); the adsorption layer (4) comprises an upper connecting ring (41), a lower connecting ring (42) and an adsorption core (43), and a tensioning device (45) is clamped between the upper connecting ring (41) and the adsorption core (43); the tensioning device (45) comprises a spring piece (451) and tensioning rubber (452), the spring piece (451) is of a circular ring structure with a V-shaped section, the tensioning rubber (452) is fixed between V-shaped openings of the spring piece (451), the top side of the spring piece (451) is fixedly bonded with the bottom side of the upper connecting ring (41), the upper end and the lower end of the adsorption core (43) are respectively fixedly bonded with the bottom end of the spring piece (451) and the top end of the lower connecting ring (42), and the upper end and the lower end of the adsorption core (43) can be tensioned by utilizing the tensioning effect of the tensioning rubber (452) on the spring piece (451); the adsorption core (43) is divided into a plurality of layers of adsorption structures with different apertures, the adsorption structures are in conical shape and are sequentially sleeved and arranged, and the conical surfaces of the adsorption structures are in forty-five-inclination arrangement; the multi-layer adsorption structure sequentially comprises a leukocyte adsorption layer, a erythrocyte adsorption layer and a platelet adsorption layer from inside to outside, wherein the leukocyte adsorption layer is of a nylon fiber structure, the adsorption aperture of the leukocyte adsorption layer is 10-20 mu m, the erythrocyte adsorption layer is of a neutral punching adsorption gum structure, the adsorption aperture of the erythrocyte adsorption layer is 7-8 mu m, the platelet adsorption layer is of a single-walled carbon nanotube non-woven membrane structure, and the adsorption aperture of the platelet adsorption layer is 2-3 mu m.
2. The pathological cell separation and adsorption extraction method according to claim 1, wherein: the conical inner wall of the adsorption structure is equally divided and fixed with a plurality of baffles (44), and the baffles (44) are perpendicular to the conical inner wall of the adsorption structure.
3. The pathological cell separation and adsorption extraction method according to claim 1, wherein: the side that goes up go-between (41) and go-between (42) and keep away from all is equipped with annular buckle (46), all be equipped with on casing (1) and cap (2) with annular buckle (46) assorted buckle groove.
4. The pathological cell separation and adsorption extraction method according to claim 1, wherein: the top of the shell cover (2) is provided with a temperature detection device (22) and a constant temperature device (23), and the detection end of the temperature detection device (22) and the output end of the constant temperature device (23) extend into the shell (1).
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Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008543326A (en) * | 2005-06-20 | 2008-12-04 | ヨン−ホ キム， | Cell separation apparatus and method |
| CN101352622A (en) * | 2008-09-17 | 2009-01-28 | 西安交通大学 | Blood separating device and separating method for filtering leucocyte |
| WO2015033457A1 (en) * | 2013-09-09 | 2015-03-12 | 株式会社日立製作所 | Cell separation and collection membrane, and culturing sheet, culturing device, and cell separation and collection method using same |
| CN104568539A (en) * | 2014-12-16 | 2015-04-29 | 何向锋 | Automatic histocyte separator |
| CN205019489U (en) * | 2015-09-30 | 2016-02-10 | 王速捷 | Tumour drops cell and adsorbs collection device |
| CN208632536U (en) * | 2018-06-25 | 2019-03-22 | 上海帛龙生物科技有限公司 | A kind of microbial cell purifier |
| JP3220702U (en) * | 2018-12-28 | 2019-03-28 | 長盛科技股▲分▼有限公司 | Automatic cell sorting equipment |
| CN209652314U (en) * | 2019-03-06 | 2019-11-19 | 成都中珠健联基因科技有限责任公司 | A kind of tumour cell adsorbent equipment of immune combination and filtering one |
| CN209669220U (en) * | 2019-01-16 | 2019-11-22 | 甘肃中医药大学 | A kind of filter separator for mesenchymal stem cells |
| CN212316092U (en) * | 2020-05-06 | 2021-01-08 | 中国医学科学院北京协和医院 | Equipment for extracting adipose-derived stem cells by negative pressure separation |
| CN112251393A (en) * | 2020-10-19 | 2021-01-22 | 北京龙迈达斯科技开发有限公司 | Cell separation method and device |
| CN112899153A (en) * | 2021-04-21 | 2021-06-04 | 江苏省疾病预防控制中心(江苏省公共卫生研究院) | Leukemia cell separation and extraction device |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20190136187A1 (en) * | 2016-06-26 | 2019-05-09 | National University Of Kaohsiung | Blood Isolation and Extraction Method and Device Thereof |
-
2021
- 2021-06-10 CN CN202110646750.6A patent/CN113174361B/en active Active
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008543326A (en) * | 2005-06-20 | 2008-12-04 | ヨン−ホ キム， | Cell separation apparatus and method |
| CN101352622A (en) * | 2008-09-17 | 2009-01-28 | 西安交通大学 | Blood separating device and separating method for filtering leucocyte |
| WO2015033457A1 (en) * | 2013-09-09 | 2015-03-12 | 株式会社日立製作所 | Cell separation and collection membrane, and culturing sheet, culturing device, and cell separation and collection method using same |
| CN104568539A (en) * | 2014-12-16 | 2015-04-29 | 何向锋 | Automatic histocyte separator |
| CN205019489U (en) * | 2015-09-30 | 2016-02-10 | 王速捷 | Tumour drops cell and adsorbs collection device |
| CN208632536U (en) * | 2018-06-25 | 2019-03-22 | 上海帛龙生物科技有限公司 | A kind of microbial cell purifier |
| JP3220702U (en) * | 2018-12-28 | 2019-03-28 | 長盛科技股▲分▼有限公司 | Automatic cell sorting equipment |
| CN209669220U (en) * | 2019-01-16 | 2019-11-22 | 甘肃中医药大学 | A kind of filter separator for mesenchymal stem cells |
| CN209652314U (en) * | 2019-03-06 | 2019-11-19 | 成都中珠健联基因科技有限责任公司 | A kind of tumour cell adsorbent equipment of immune combination and filtering one |
| CN212316092U (en) * | 2020-05-06 | 2021-01-08 | 中国医学科学院北京协和医院 | Equipment for extracting adipose-derived stem cells by negative pressure separation |
| CN112251393A (en) * | 2020-10-19 | 2021-01-22 | 北京龙迈达斯科技开发有限公司 | Cell separation method and device |
| CN112899153A (en) * | 2021-04-21 | 2021-06-04 | 江苏省疾病预防控制中心(江苏省公共卫生研究院) | Leukemia cell separation and extraction device |
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