CN115992096A - Tumor cell detection method - Google Patents
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- CN115992096A CN115992096A CN202310150273.3A CN202310150273A CN115992096A CN 115992096 A CN115992096 A CN 115992096A CN 202310150273 A CN202310150273 A CN 202310150273A CN 115992096 A CN115992096 A CN 115992096A
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Abstract
The invention relates to a tumor cell detection method, which relates to the technical field of cell detection and comprises the steps of preparing a cell suspension, polymerizing, separating, detecting and the like. The present application enables "clumps" of tumor cells to be broken up into individual cells for later counting of the number of circulating tumor cells using a flow cytometer. And the tumor cells of the 'group' are not directly contacted when being dispersed, the probability of damaging various cells is reduced, the integrity of the cells is maintained, and the counting in a flow cytometer is convenient.
Description
Technical Field
The invention relates to the technical field of cell detection, in particular to a tumor cell detection method.
Background
Cancer is one of the most serious diseases which endanger human health at present, and detection of cancer cells has important significance both in the diagnosis and treatment of cancer. Circulating tumor cells (CTCs for short) are tumor cells that shed from a tumor and enter human peripheral blood; clinically, whether the tumor exists or not can be assisted by detecting whether circulating tumor cells exist in human peripheral blood, and the effect of tumor treatment can be judged by monitoring the content of the circulating tumor cells in human peripheral blood.
In the process of treating tumors, as the medical staff has already known the types of tumor cells, the number of circulating tumor cells in the blood can be detected by the flow cytometer by taking blood and then preparing a cell suspension and then placing the cell suspension on the flow cytometer. The doctor can know how the treatment effect is achieved through the increase and decrease of the number of the circulating tumor cells in the blood, and then the doctor is assisted in judging how to conduct the next stage of diagnosis and treatment.
With respect to the above related art, the inventor believes that, since the circulating tumor cells exist not only in the form of single cells but also in the form of "groups" composed of a plurality of cells in the peripheral blood of the human body, the tumor cells in the form of "groups" may not only affect the flow cytometer to determine the type of the tumor cells, but also affect the flow cytometer to count the number of the tumor cells, thereby being unfavorable for the doctor to determine the disease condition.
Disclosure of Invention
In order to disperse the tumor cells of the 'group', and further facilitate the counting of the circulating tumor cells, the invention provides a tumor cell detection method.
The invention provides a tumor cell detection method, which adopts the following technical scheme:
a method of tumor cell detection comprising the steps of:
preparation of cell suspensions: preparing the collected blood into a cell suspension;
polymerization: the magnetic particles coated with the first specific antibody are put into a cell suspension, so that the first specific antibody is combined with tumor cells;
separating: treating the cell suspension with a separation device to separate the tumor cells of the "clusters";
and (3) detection: placing the cell suspension into a flow cytometer for detection, and counting the circulating tumor cells;
the separating device comprises a base, a liquid containing pipe, a magnetic field generator and a stirring mechanism, wherein the liquid containing pipe is arranged on the base, the stirring mechanism is arranged on the base, the output end of the stirring mechanism is arranged in the liquid containing pipe, and the magnetic field emitter is arranged on the base and outside the liquid containing pipe;
in the separation step, the cell suspension is put into a liquid containing tube, and under the stirring action of a stirring mechanism, the cell suspension spins in the liquid containing tube, then a magnetic field generator is started, and a magnetic field is formed in the liquid containing tube, so that the magnetic force born by the magnetic particles is opposite to the spinning direction of the cell suspension in the liquid containing tube.
By adopting the technical scheme, in the polymerization step, the first specific antibody coated on the magnetic particles can be combined with tumor cells, and as part of tumor cells exist in a form of 'groups', a plurality of magnetic particles can be combined on the 'groups' of tumor cells; in the separation step, the cell suspension spins in the liquid containing tube, when the magnetic field generator acts, the magnetic particles are stressed in the opposite direction to the moving direction of the magnetic particles, and the 'group' tumor cells continue to spin under the action of inertia, so that the 'group' tumor cells are pulled and dispersed, and then the number of circulating tumor cells is counted by using a flow cytometer. The tumor cells of the 'group' are not directly contacted when dispersed, the probability of breakage of various cells is reduced, the integrity of the cells is maintained, and the counting in a flow cytometer is facilitated.
Optionally, the stirring mechanism includes driving motor, rolling disc and puddler, driving motor sets up on the base, the rolling disc sets up on driving motor's the output shaft, the puddler becomes the heart setting and is in on the rolling disc.
Through adopting above-mentioned technical scheme, when stirring cell suspension, stretch into the puddler in the flourishing liquid pipe to make rolling disc and flourishing liquid pipe coaxial, later driving motor drive carousel rotates, the puddler alright stir cell suspension. Because the stirring rod is used for stirring the cell suspension, cells in the cell suspension are not easy to be subjected to shearing force, and the probability of cell breakage is further reduced.
Optionally, the separating device further comprises a lifting mechanism, the lifting mechanism comprises a lifting frame and an electric pushing cylinder, a cylinder body of the electric pushing cylinder is arranged on the base, the lifting frame is arranged on a piston rod of the electric pushing cylinder, and the driving motor is arranged on the lifting frame;
the lifting frame is further provided with a positioning mechanism, the positioning mechanism comprises a positioning block, the positioning block is arranged on the lifting frame, the diameter of the positioning block, which is close to one end of the lifting frame, is larger than that of the other end of the positioning block, and one end, which is far away from the lifting frame, of the positioning block stretches into the liquid containing pipe.
By adopting the technical scheme, before stirring the cell suspension, the positioning block stretches into the liquid containing pipe, so that the liquid containing pipe is positioned, and the end part of the liquid containing pipe is plugged; when stirring the cell suspension, the liquid containing pipe is not easy to shake, so that the cell suspension is not easy to be poured, the number of the circulating tumor cells is not easy to change, and the detection precision is maintained.
Optionally, the locating piece joint is in on the crane, the rotating disk rotates to set up on the locating piece, just the rotating disk with driving motor joint.
By adopting the technical scheme, before stirring the new cell suspension, the new positioning block is clamped on the lifting frame, and the rotating disc can be clamped with the driving motor at the moment; after stirring, the positioning block is detached from the lifting frame, so that the rotating disc and the stirring rod can be detached, the stirring rod is convenient to replace, and the probability of pollution of the cell suspension is reduced.
Optionally, a standing step is further arranged between the separation step and the detection step,
standing: standing the cell suspension in a liquid containing tube for a period of time, so that the first specific antibody is combined with tumor cells again;
after the completion of the standing step, the separation and standing steps are repeated a plurality of times.
By adopting the technical scheme, after one-time separation is finished, the cell suspension is kept stand in the liquid containing tube for a period of time, at the moment, small 'groups' of tumor cells which are just dispersed can be combined with the first specific antibody again, and then the 'groups' of the tumor cells are separated again under the action of a magnetic field; the tumor cell "clusters" separated many times eventually become individual tumor cells, which in turn facilitate counting in a flow cytometer.
Optionally, a second polymerization step is further arranged between the standing step and the detection step;
second polymerization: the second specific antibody labeled with fluorescein is administered to the cell suspension, and the second specific antibody is allowed to bind to tumor cells.
Because the content of the circulating tumor cells in the peripheral blood of the human body is small, the flow cytometer is easy to have false recognition phenomenon when recognizing the circulating tumor cells, so that the number of the detected circulating tumor cells has deviation; by adopting the technical scheme, the flow cytometer can simultaneously identify the magnetic particles and the second specific antibody marked by the fluorescein when identifying the circulating tumor cells, so that the probability of identifying the circulating tumor cells is greatly increased, and the probability of 'missed detection' and 'false detection' is reduced.
In summary, the present invention includes at least one of the following beneficial technical effects:
1. through the setting of the separation step, the cell suspension spins in the liquid containing tube, and the magnetic particles are subjected to the force opposite to the moving direction of the magnetic particles under the action of the magnetic field generator, so that the 'group' tumor cells are pulled and dispersed into single cells, and then the number of circulating tumor cells is counted by using a flow cytometer. The tumor cells of the 'group' are not directly contacted when dispersed, the probability of breakage of various cells is reduced, the integrity of the cells is maintained, and the counting in a flow cytometer is facilitated.
2. Through the setting of puddler, when stirring cell suspension, the puddler is difficult for applying to the cell in the cell suspension with shearing force, when making the cell suspension spin in holding the liquid pipe, has reduced the probability of cell damage.
3. After the primary separation is completed, the tumor cell clusters may not be completely separated, and the tumor cell clusters can be combined with the first specific antibody again through setting a standing step, repeating the steps of separation and standing after standing for a plurality of times, and then separating the tumor cell 'clusters' through the action of a magnetic field; thereby the tumor cells are separated more thoroughly.
4. Because the content of the circulating tumor cells in the peripheral blood of the human body is small, the flow cytometer is easy to have false recognition phenomenon when recognizing the circulating tumor cells, so that the number of the detected circulating tumor cells has deviation; through the setting of the second polymerization step, the circulating tumor cells are indirectly marked with fluorescein, so that the probability of being identified by the circulating tumor cells is greatly increased, and the probability of 'missed detection' and 'false detection' is reduced.
Drawings
FIG. 1 is a schematic flow diagram of an embodiment of the present application;
FIG. 2 is a schematic view of the overall structure of a separation device according to an embodiment of the present application;
FIG. 3 isbase:Sub>A schematic cross-sectional view of A-A of FIG. 2;
fig. 4 is an enlarged schematic view of a portion B in fig. 3.
Reference numerals illustrate: 100. a base; 110. a placement cavity; 120. a liquid containing tube; 200. a lifting mechanism; 210. a lifting frame; 211. a guide hole; 220. an electric pushing cylinder; 300. a positioning mechanism; 310. a positioning block; 320. a guide rod; 330. a locking assembly; 331. a locking block; 3311. a guide surface; 332. a compression spring; 400. a stirring mechanism; 410. a driving motor; 420. a first end face toothed disc; 430. a second end face fluted disc; 440. a rotating disc; 450. a stirring rod; 500. a magnetic field generator.
Detailed Description
The invention is described in further detail below in connection with fig. 1-4.
The embodiment of the application discloses a tumor cell detection method, referring to fig. 1, the tumor cell detection method comprises the following steps:
s1, preparing a cell suspension: preparing collected blood into a cell suspension which can enter a flow cytometer for detection;
s2, polymerization: the magnetic particles coated with the first specific antibody are put into a cell suspension, and then the magnetic particles are oscillated and kept stand for a period of time, so that the first specific antibody and tumor cells generate antigen-antibody reaction, and then the first specific antibody and the tumor cells generate specific binding;
s3, separating: treating the cell suspension with a separation device to separate the tumor cells of the "clusters";
s4, standing: standing the cell suspension in the liquid containing tube 120 for a period of time to enable the first specific antibody to react with the tumor cells again in an antigen-antibody reaction mode, and further enable the first specific antibody to be combined with the tumor cells again in a specific mode;
repeating the steps of S3 separation and S4 standing for a plurality of times after the standing step is finished, so that the tumor cells of the 'group' are separated more thoroughly;
s5, second polymerization: adding a second specific antibody marked by fluorescein into the cell suspension, and then vibrating and standing for a period of time to enable the second specific antibody to react with the antigen and the antibody of the tumor cell, so that the second specific antibody and the tumor cell are specifically combined;
s6, detecting, namely placing the cell suspension into a flow cytometer for detection, and detecting the cells which are attached with magnetic particles and combined with the second specific antibody, so as to finally obtain the number of the circulating tumor cells.
Since the "clustered" tumor cells are split into individual tumor cells prior to counting the circulating tumor cells, and the individual tumor cells are antigen-antibody reacted with the second specific antibody; in the detection step, the cells attached with the magnetic particles and combined with the second specific antibody can be identified in a targeted manner, so that the probability of missed detection and false detection of tumor cells is reduced, and the detection precision is improved.
Referring to fig. 2 to 4, in the S3 separation step, a separation device is used, which includes a base 100, a liquid-holding tube 120 for holding a cell suspension, a stirring mechanism 400 for stirring the suspension, a lifting mechanism 200 for driving the stirring mechanism 400 to lift, and a magnetic field generator 500 for splitting a "group" of tumor cells. The base 100 is provided with a placing cavity 110 for placing the liquid containing tube 120, and the liquid containing tube 120 is placed in the placing cavity 110.
Referring to fig. 2 to 4, the lifting mechanism 200 includes a lifting frame 210 and an electric push cylinder 220, a cylinder body of the electric push cylinder 220 is fixedly connected to the base 100 through a bolt, the lifting frame 210 is fixedly connected to a piston rod of the electric push cylinder 220 through a bolt, and the electric push cylinder 220 is vertically arranged.
Referring to fig. 2 to 4, a positioning mechanism 300 is detachably coupled to the lifting frame 210, and the positioning mechanism 300 includes a positioning block 310, a plurality of guide bars 320, and locking assemblies 330 in the same number as the guide bars 320. The locking assembly 330 comprises a compression spring 332 and a locking block 331, wherein the locking block 331 is slidably arranged on the lifting frame 210, one end of the compression spring 332 is abutted against the lifting frame 210, and the other end is abutted against the locking block 331. The lifting frame 210 is provided with a guide hole 211 for the guide rod 320 to pass through, and the guide rod 320 is arranged in the guide hole 211 in a penetrating way; when the guide rod 320 is inserted into the guide hole 211, the locking block 331 is abutted against the guide rod 320 by the elastic force of the compression spring 332. A guide surface 3311 is formed between an end surface of the locking block 331 close to the guide rod 320 and an end surface close to the base 100, the guide rod 320 is firstly abutted against the guide surface 3311 when being conveyed into the guide hole 211, and under the guide action of the guide surface 3311, the locking block 331 presses the compression spring 332, so that an end surface of the locking block 331 far from the compression spring 332 is abutted against the guide rod 320. The positioning block 310 is in a truncated cone shape, and the diameter of one end of the positioning block 310, which is close to the lifting frame 210, is larger than that of the other end.
Referring to fig. 2 to 4, the stirring mechanism 400 includes a driving motor 410, a first end face fluted disc 420, a second end face fluted disc 430, a rotating disc 440, and a stirring rod 450, wherein the driving motor 410 is fixedly connected to the lifting frame 210 through bolts, and the first end face fluted disc 420 is coaxially connected to an output shaft of the driving motor 410 through keys; the rotating disc 440 is rotatably connected to the positioning hole, and the second end face fluted disc 430 is coaxially and fixedly connected to the rotating disc 440. When the guide rod 320 is completely inserted into the guide hole 211, the first end face gear 420 is engaged with the second end face gear 430, and the driving motor 410 can drive the rotating disc 440 to rotate through the first end face gear 420 and the second end face gear 430. The stirring rod 450 is screw-coupled to the rotating disk 440, and the stirring rod 450 is eccentrically installed to the rotating disk 440. The magnetic field generator 500 is fixedly connected to the base 100 by bolts, and the magnetic field generator 500 is disposed at both sides of the liquid containing tube 120.
When the separation step is performed, the cell suspension is put into the liquid containing tube 120, then the liquid containing tube 120 is embedded into the placing cavity 110, and then the electric pushing cylinder 220 drives the lifting frame 210 to move downwards until one end of the positioning block 310, which is far away from the lifting frame 210, is inserted into the opening of the liquid containing tube 120, and at this time, the fixing and sealing of the liquid containing tube 120 by the positioning block 310 are realized. Then, the motor 410 is driven to rotate the stirring rod 450, and the stirring rod 450 stirs the cell suspension to spin the cell suspension in the liquid containing tube 120; then the magnetic field generator 500 is electrified, a magnetic field is formed in the liquid containing tube 120 under the action of the magnetic field generator 500, and the direction of the magnetic field applied to the magnetic particles is opposite to the moving direction of the magnetic particles in the liquid containing tube 120. The tumor cells on the surface of the tumor cell group can be separated, and after multiple times of separation, the tumor cell group can be separated into single tumor cells.
When the separation step is performed, the cells in the cell suspension are not easily subjected to shearing force, so that the cells in the cell suspension are not easily damaged, and when the cell suspension is detected by using the flow cytometer, the flow cytometer can count various cells more accurately. Under the condition of more accurate counting, the accuracy of the doctor on disease diagnosis is improved.
When detecting different cell suspensions, a detector can directly put a new liquid containing tube 120 in the placing cavity 110, and the detector can detach the positioning block 310 to directly replace the stirring rod 450, so that the probability of pollution of the cell suspension is reduced.
Comparative example
Comparative example 1: the comparative example uses a flow cytometer to detect the blood which is not subjected to the separation step, and the clinical diagnosis result of the patient is lung cancer, and the blood collection amount is 3ml.
The blood is treated as follows:
s1: preparation of cell suspensions: preparing the collected blood into a cell suspension;
s2: polymerization: labeling tumor cells with a fluorescent-labeled specific antibody;
s3: and (3) detection: the cell suspension was placed into a flow cytometer for detection and the circulating tumor cells were counted.
The results of the blood test are shown in Table 1.
TABLE 1
| Detecting items | Results | Unit (B) | Reference range |
| White blood cell count | 1.06 | 10^9/L | 4.00-10.00 |
| Neutrophil count | 0.59 | 10^9/L | 2.00-7.00 |
| Lymphocyte number | 0.46 | 10^9/L | 0.80-4.00 |
| Monocyte number | 0.01 | 10^9/L | 0.12-1.20 |
| Eosinophil number | 0.00 | 10^9/L | 0.02-0.50 |
| Basophil number | 0.00 | 10^9/L | 0.00-0.10 |
| Number of red blood cells | 1.99 | 10^12/L | 3.50-5.50 |
| Platelet count | 17 | 10^9/L | 100-300 |
| Rare cell number | 8 | Personal (S) | |
| Number of suspected rare cell clusters | 4 | Personal (S) |
Comparative example 2: the comparative example uses a flow cytometer to detect the blood which is not subjected to the separation step, and the clinical diagnosis result of the patient is lung cancer, and the blood collection amount is 3ml.
The blood is treated as follows:
s1: preparation of cell suspensions: preparing the collected blood into a cell suspension;
s2: polymerization: labeling tumor cells with a fluorescent-labeled specific antibody;
s3: and (3) detection: the cell suspension was placed into a flow cytometer for detection and the circulating tumor cells were counted.
The test results are shown in Table 2.
TABLE 2
| Detecting items | Results | Unit (B) | Reference rangeSurrounding wall |
| White blood cell count | 4.09 | 10^9/L | 4.00-10.00 |
| Neutrophil count | 2.15 | 10^9/L | 2.00-7.00 |
| Lymphocyte number | 1.56 | 10^9/L | 0.80-4.00 |
| Monocyte number | 0.34 | 10^9/L | 0.12-1.20 |
| Eosinophil number | 0.04 | 10^9/L | 0.02-0.50 |
| Basophil number | 0.00 | 10^9/L | 0.00-0.10 |
| Number of red blood cells | 4.21 | 10^12/L | 3.50-5.50 |
| Platelet count | 71 | 10^9/L | 100-300 |
| Rare cell number | 11 | Personal (S) | |
| Number of suspected rare cell clusters | 2 | Personal (S) |
Examples
Example 1: the present comparative example uses a flow cytometer to detect the blood subjected to the separation step, and the patient is the same patient as that in comparative example 1, the blood collection amount is 3ml, and the blood collection lot is the same lot as that in comparative example 1.
The treatment steps of the blood are tumor cell detection methods provided in the embodiments of the present application.
The test results are shown in Table 3.
TABLE 3 Table 3
| Detecting items | Results | Unit (B) | Reference range |
| White blood cell count | 1.05 | 10^9/L | 4.00-10.00 |
| Neutrophil count | 0.59 | 10^9/L | 2.00-7.00 |
| Lymphocyte number | 0.46 | 10^9/L | 0.80-4.00 |
| Monocyte number | 0.01 | 10^9/L | 0.12-1.20 |
| Eosinophil number | 0.00 | 10^9/L | 0.02-0.50 |
| Basophil number | 0.00 | 10^9/L | 0.00-0.10 |
| Number of red blood cells | 2.01 | 10^12/L | 3.50-5.50 |
| Platelet count | 17 | 10^9/L | 100-300 |
| Rare cell number | 21 | Personal (S) | |
| Number of suspected rare cell clusters | 1 | Personal (S) |
Example 2: the present comparative example uses a flow cytometer to detect the blood subjected to the separation step, and the patient is the same patient as that in comparative example 2, the blood collection amount is 3ml, and the blood collection lot is the same lot as that in comparative example 2.
The treatment steps of the blood are tumor cell detection methods provided in the embodiments of the present application.
The test results are shown in Table 4.
TABLE 4 Table 4
| Detecting items | Results | Unit (B) | Reference range |
| White blood cell count | 4.10 | 10^9/L | 4.00-10.00 |
| Neutrophil count | 2.15 | 10^9/L | 2.00-7.00 |
| Lymphocyte number | 1.55 | 10^9/L | 0.80-4.00 |
| Monocyte number | 0.34 | 10^9/L | 0.12-1.20 |
| Eosinophil number | 0.04 | 10^9/L | 0.02-0.50 |
| Basophil number | 0.00 | 10^9/L | 0.00-0.10 |
| Number of red blood cells | 4.21 | 10^12/L | 3.50-5.50 |
| Platelet count | 71 | 10^9/L | 100-300 |
| Rare cell number | 18 | Personal (S) | |
| Number of suspected rare cell clusters | 0 | Personal (S) |
The rare cells in comparative example 1, comparative example 2 and example 2 are tumor cells to be detected. It can be seen from the combination of comparative example 1 and example 1 or the combination of comparative example 2 and example 2 that the number of rare cell clusters is reduced and the number of rare cells is increased by splitting the tumor cell "clusters" by using the separation step, thereby proving that the separation step can effectively split the tumor cell clusters. While the number of other cells was substantially unchanged, demonstrating that the cells were not substantially destroyed when the isolation step was performed.
The implementation principle of the tumor cell detection method in the embodiment of the application is as follows:
before counting and detecting the circulating tumor cells, splitting the tumor cell 'group', so that the tumor cells of the 'group' are split into single tumor cells, and the single tumor cells and the second specific antibody perform antigen-antibody reaction, thereby indirectly marking the tumor cells; in the detection step, the cells attached with the magnetic particles and combined with the second specific antibody can be identified in a targeted manner, so that the probability of missed detection and false detection of tumor cells is reduced, and the detection precision is improved.
When dividing the tumor cells of the group, the cells in the cell suspension are not easy to be subjected to shearing force, so that the cells in the cell suspension are not easy to be damaged, and when detecting the cell suspension by using a flow cytometer, the flow cytometer can count various cells more accurately. Under the condition of more accurate counting, the accuracy of the doctor on disease diagnosis is improved.
The above embodiments are not intended to limit the scope of the present invention, so: all equivalent changes in structure, shape and principle of the invention should be covered in the scope of protection of the invention.
Claims (6)
1. A method for detecting tumor cells, comprising the steps of:
preparation of cell suspensions: preparing the collected blood into a cell suspension;
polymerization: the magnetic particles coated with the first specific antibody are put into a cell suspension, so that the first specific antibody is combined with tumor cells;
separating: treating the cell suspension with a separation device to separate the tumor cells of the "clusters";
and (3) detection: placing the cell suspension into a flow cytometer for detection, and counting the circulating tumor cells;
the separating device comprises a base (100), a liquid containing pipe (120), a magnetic field generator (500) and a stirring mechanism (400), wherein the liquid containing pipe (120) is arranged on the base (100), the stirring mechanism (400) is arranged on the base (100), the output end of the stirring mechanism (400) is arranged in the liquid containing pipe (120), and the magnetic field transmitter is arranged on the base (100) and is arranged outside the liquid containing pipe (120);
in the separation step, the cell suspension is put into the liquid containing tube (120), and under the stirring action of the stirring mechanism (400), the cell suspension spins in the liquid containing tube (120), and then the magnetic field generator (500) is started, and a magnetic field is formed in the liquid containing tube (120), so that the magnetic force exerted by the magnetic particles is opposite to the spinning direction of the cell suspension in the liquid containing tube (120).
2. The method for detecting tumor cells according to claim 1, wherein: the stirring mechanism (400) comprises a driving motor (410), a rotating disc (440) and a stirring rod (450), wherein the driving motor (410) is arranged on the base (100), the rotating disc (440) is arranged on an output shaft of the driving motor (410), and the stirring rod (450) is arranged on the rotating disc (440) in a variable center mode.
3. The method for detecting tumor cells according to claim 2, wherein: the separating device further comprises a lifting mechanism (200), the lifting mechanism (200) comprises a lifting frame (210) and an electric pushing cylinder (220), a cylinder body of the electric pushing cylinder (220) is arranged on the base (100), the lifting frame (210) is arranged on a piston rod of the electric pushing cylinder (220), and the driving motor (410) is arranged on the lifting frame (210);
the lifting frame (210) is further provided with a positioning mechanism (300), the positioning mechanism (300) comprises a positioning block (310), the positioning block (310) is arranged on the lifting frame (210), the diameter of the positioning block (310) close to one end of the lifting frame (210) is larger than that of the other end, and one end, away from the lifting frame (210), of the positioning block (310) stretches into the liquid containing tube (120).
4. A method of detecting tumor cells according to claim 3, wherein: the positioning block (310) is clamped on the lifting frame (210), the rotating disc (440) is rotatably arranged on the positioning block (310), and the rotating disc (440) is clamped with the driving motor (410).
5. The method for detecting tumor cells according to any one of claims 1 to 4, wherein: a standing step is also arranged between the separation step and the detection step,
standing: standing the cell suspension in a liquid containing tube (120) for a period of time, so that the first specific antibody is combined with the tumor cells again;
after the completion of the standing step, the separation and standing steps are repeated a plurality of times.
6. The method for detecting tumor cells according to claim 5, wherein: a second polymerization step is further arranged between the standing step and the detection step;
second polymerization: the second specific antibody labeled with fluorescein is administered to the cell suspension, and the second specific antibody is allowed to bind to tumor cells.
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| US20160083781A1 (en) * | 2007-10-31 | 2016-03-24 | Akonni Biosystems, Inc. | Integrated sample processing system |
| CN106520543A (en) * | 2016-10-09 | 2017-03-22 | 陈静 | High-throughput quick capturing and purifying apparatus and method for circulating tumor cells |
| CN108982839A (en) * | 2018-06-12 | 2018-12-11 | 广州四叶草健康科技有限公司 | Circulating tumor cell detection method based on immunomagnetic beads and flow cytometer |
| CN213467474U (en) * | 2020-10-20 | 2021-06-18 | 杭州富阳富春涂装有限公司 | Coating stirring device |
| CN215404280U (en) * | 2020-11-11 | 2022-01-04 | 贵州医科大学附属医院 | Tumor cell culture device |
| CN214830401U (en) * | 2021-03-02 | 2021-11-23 | 上海青赛生物科技有限公司 | Primary cell dispersion devices |
| CN115287249A (en) * | 2022-07-07 | 2022-11-04 | 中国医学科学院医学生物学研究所 | Separation device and separation method for primary cells of cynomolgus monkey kidney |
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