CN115201476A - Circulating tumor cell detection equipment and use method thereof - Google Patents
Circulating tumor cell detection equipment and use method thereof Download PDFInfo
- Publication number
- CN115201476A CN115201476A CN202211120006.3A CN202211120006A CN115201476A CN 115201476 A CN115201476 A CN 115201476A CN 202211120006 A CN202211120006 A CN 202211120006A CN 115201476 A CN115201476 A CN 115201476A
- Authority
- CN
- China
- Prior art keywords
- chamber
- gun
- liquid
- moves
- cell chip
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000001514 detection method Methods 0.000 title claims abstract description 50
- 208000005443 Circulating Neoplastic Cells Diseases 0.000 title claims abstract description 31
- 238000000034 method Methods 0.000 title claims abstract description 20
- 239000007788 liquid Substances 0.000 claims abstract description 172
- 238000006243 chemical reaction Methods 0.000 claims abstract description 93
- 210000004027 cell Anatomy 0.000 claims abstract description 85
- 239000000523 sample Substances 0.000 claims abstract description 78
- 238000012546 transfer Methods 0.000 claims abstract description 62
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 53
- 238000012360 testing method Methods 0.000 claims abstract description 39
- 210000002966 serum Anatomy 0.000 claims abstract description 21
- 238000009396 hybridization Methods 0.000 claims abstract description 18
- 238000004043 dyeing Methods 0.000 claims abstract description 15
- 210000004881 tumor cell Anatomy 0.000 claims abstract description 14
- 239000002699 waste material Substances 0.000 claims description 62
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 52
- 238000004140 cleaning Methods 0.000 claims description 34
- 238000005406 washing Methods 0.000 claims description 13
- 238000010438 heat treatment Methods 0.000 claims description 11
- 239000003086 colorant Substances 0.000 claims description 7
- 238000002347 injection Methods 0.000 claims description 6
- 239000007924 injection Substances 0.000 claims description 6
- 238000005192 partition Methods 0.000 claims description 6
- 238000007664 blowing Methods 0.000 claims description 4
- 238000007789 sealing Methods 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims 1
- 210000000349 chromosome Anatomy 0.000 abstract description 2
- 238000012744 immunostaining Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 9
- 206010028980 Neoplasm Diseases 0.000 description 8
- 206010027476 Metastases Diseases 0.000 description 4
- 210000005259 peripheral blood Anatomy 0.000 description 4
- 239000011886 peripheral blood Substances 0.000 description 4
- 239000011324 bead Substances 0.000 description 3
- 238000003745 diagnosis Methods 0.000 description 3
- 230000009401 metastasis Effects 0.000 description 3
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 238000010191 image analysis Methods 0.000 description 2
- 238000011528 liquid biopsy Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 208000037323 Rare tumor Diseases 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000001640 apoptogenic effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000012864 cross contamination Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001839 endoscopy Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000003125 immunofluorescent labeling Methods 0.000 description 1
- 238000007901 in situ hybridization Methods 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000012474 protein marker Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000001360 synchronised effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 239000000439 tumor marker Substances 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56966—Animal cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6841—In situ hybridisation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N1/31—Apparatus therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Hematology (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Organic Chemistry (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to the field of cell detection, in particular to circulating tumor cell detection equipment and a using method thereof, wherein the detection equipment comprises a detection frame and a plurality of sample test tubes which are centrifugally separated by a centrifugal machine, each sample test tube is filled with serum containing tumor cells, the detection frame is provided with a sample chamber, a consumable chamber and a reaction chamber, the plurality of sample test tubes are placed in the sample chamber, a plurality of cell chips are placed in the reaction chamber, the serum in each sample test tube is injected into the corresponding cell chip through a liquid transfer gun arranged on the detection frame, and a plurality of reagents in the consumable chamber are sequentially dripped onto the corresponding cell chip through the liquid transfer gun so as to dye and perform hybridization reaction on the tumor cells on the cell chip. The invention automatically drips reagents on a plurality of cell chips in the reaction chamber through the pipette to carry out dyeing and hybridization reaction, thereby not only greatly improving the detection efficiency, but also reducing false negative through a chromosome probe/immunostaining identification method.
Description
Technical Field
The invention relates to the field of cell detection, in particular to circulating tumor cell detection equipment and a using method thereof.
Background
Diagnosis, efficacy assessment and recurrence monitoring of tumors are important tools for tumor therapy. The liquid biopsy obtains the tumor cells and the molecular information thereof in a non-invasive sampling mode, and has obvious advantages in sensitivity and specificity compared with the traditional tumor marker detection, imaging detection and endoscopy. Circulating Tumor Cells (CTCs) are derived from tumor primary foci or tumor cells released from solid tumors or metastases into peripheral blood circulation due to diagnosis and treatment, and have very small number of CTCs in peripheral blood, accounting for only 1/10 of white blood cells in peripheral blood 6 -1/10 7 Therefore, they are also called rare tumor cells. Most of CTCs are apoptotic or phagocytized after entering peripheral blood, and a small amount of CTCs with high vitality and metastasis potential can survive in the circulatory system, and when meeting the matrix environment of appropriate organs and tissues, tumor metastasis occurs, which is an important reason for postoperative recurrence and distant metastasis of malignant tumor patients and also an important factor for death of tumor patients. The effective detection of circulating tumor cells by liquid biopsy provides an important means for tumor diagnosis and treatment.
At present, the following methods are mainly adopted for detecting circulating tumor cells by adopting detection equipment: 1. the filtration method (also called ISET), which separates CTC using a 8 μm sieve according to cell size, has disadvantages in that tumor cell size and blood-derived cells overlap and are difficult to distinguish, and false negative is high. 2. An immune antigen-antibody sorting method, which aims at the immune magnetic bead of a protein marker (antigen) on the surface of a CTC cell membrane to positively capture CTC, because the expression of the protein (antigen) on the surface of the CTC cell membrane is dynamic, the antibody can not capture the CTC under the condition that the surface protein (target) is not expressed, and the false negative is high.
Disclosure of Invention
In order to solve the technical problems, the invention provides a circulating tumor cell detection device which can improve the detection efficiency and reduce the false negative.
The technical scheme adopted by the invention for solving the technical problems is as follows: a circulating tumor cell detection device comprises a detection frame and a plurality of sample test tubes which are centrifugally separated by a centrifugal machine, wherein serum containing tumor cells is filled in each sample test tube, the detection frame is provided with a sample chamber, a consumable chamber and a reaction chamber, the plurality of sample test tubes are placed in the sample chamber, a plurality of cell chips are placed in the reaction chamber, the serum in each sample test tube is injected into the corresponding cell chip through a liquid transfer gun arranged on the detection frame, and then a plurality of reagents in the consumable chamber are dripped onto the corresponding cell chip in sequence through the liquid transfer gun to enable the tumor cells on the cell chip to be dyed and hybridized; the indoor stationary liquid room that is equipped with of consumptive material, washing liquid room, ethanol room, probe reagent room, dye reagent room and demonstration liquid room, after the cell chip was injected into to the serum, the pipetting gun is dripped indoor stationary liquid, the indoor washing liquid of washing liquid, the indoor ethanol of ethanol, the indoor probe reagent of probe reagent and the indoor dye reagent of stationary liquid in proper order on this cell chip in order to carry out dyeing and hybridization reaction, and the reaction is accomplished the back, the rethread the pipetting gun is dripped indoor washing liquid of washing liquid and the indoor demonstration liquid of demonstration liquid in proper order on the cell chip in order to wash and show.
Preferably, a gun nozzle chamber is arranged in the consumable chamber, a plurality of gun nozzles matched with the liquid-transferring gun are placed in the gun nozzle chamber, and the liquid-transferring gun moves to the gun nozzle chamber to automatically replace new gun nozzles before liquid is absorbed by the liquid-transferring gun at each time.
Preferably, a waste treatment chamber is arranged on the detection frame, and a gun nozzle of the pipetting gun automatically falls off to the waste treatment chamber after pipetting is completed.
Preferably, the waste treatment chamber is a closed space surrounded by a plurality of partition boards, an opening is formed in the partition board on the upper side, and the fallen gun nozzle enters the closed space through the opening.
Preferably, the detection frame is provided with a guide rail positioned above the sample chamber, the fixed liquid chamber, the cleaning liquid chamber, the ethanol chamber, the probe reagent chamber, the dye reagent chamber, the display liquid chamber, the gun nozzle chamber, the reaction chamber and the waste treatment chamber, the guide rail is provided with an automatic guide trolley, and the automatic guide trolley drives the liquid-transferring gun arranged on the automatic guide trolley to move along the guide rail.
Preferably, the reaction chamber is a constant-temperature closed bin, an automatic bin gate is arranged on the constant-temperature closed bin, when the automatic guide trolley moves to the position of the automatic bin gate, the automatic bin gate is automatically opened, the automatic guide trolley drives the liquid-transferring gun to enter the constant-temperature closed bin along the guide rail, and the liquid-transferring gun drips the corresponding reagent on the corresponding cell chip in the constant-temperature closed bin.
Preferably, the constant-temperature closed bin is divided into an upper chamber and a lower chamber by a bedplate, a liquid injection platform is arranged in the upper chamber on the bedplate, a plurality of cell chips are arranged on the liquid injection platform, and a heating module capable of heating and insulating the upper chamber is arranged in the lower chamber on the lower side of the bedplate.
Preferably, a test tube rack is arranged in the sample chamber, and a plurality of sample test tubes are inserted on the test tube rack.
The invention also provides a using method of the circulating tumor cell detection equipment, which comprises the following steps:
(1) Placing the centrifugally separated sample test tube in a sample chamber, moving a liquid transfer gun to a gun nozzle chamber to automatically install a gun nozzle, then moving the sample chamber to absorb serum in the sample test tube and moving the serum to a reaction chamber, then injecting the serum onto a cell chip in the reaction chamber, then moving the liquid transfer gun to a waste treatment chamber, and automatically dropping the gun nozzle into the waste treatment chamber;
(2) The liquid transferring gun moves to a gun nozzle chamber to install a new gun nozzle, then moves to a stationary liquid chamber to absorb stationary liquid and moves to a reaction chamber, the stationary liquid is dripped onto a cell chip in the reaction chamber, then the liquid transferring gun moves to a waste treatment chamber, and the gun nozzle automatically falls off to the waste treatment chamber; then the pipetting gun moves back to the reaction chamber, after the cell chip is incubated for a period of time, air is blown out through the pipetting gun to blow dry the cell chip;
(3) Moving a liquid transfer gun to a gun nozzle chamber to install a new gun nozzle, then moving the liquid transfer gun to a cleaning chamber to absorb cleaning liquid and moving the cleaning liquid to a reaction chamber, wherein the liquid transfer gun firstly drips one part of the cleaning liquid on the cell chip, and drips the other part of the cleaning liquid after a period of time; then the liquid transferring gun is moved to a waste treatment chamber, and the gun nozzle automatically falls off into the waste treatment chamber;
(4) Moving the liquid transfer gun to a gun nozzle chamber to install a new gun nozzle, then moving the liquid transfer gun to an ethanol chamber to absorb ethanol and moving the liquid transfer gun to a reaction chamber, dripping one part of ethanol to the cell chip by the liquid transfer gun, and dripping the other part of ethanol after a period of time; moving the liquid transfer gun to a waste treatment chamber, automatically dropping a gun nozzle to the waste treatment chamber, moving the liquid transfer gun back to the reaction chamber, and blowing air through the liquid transfer gun to blow dry the cell chip after the cell chip is incubated for a period of time;
(5) The pipetting gun moves to a gun nozzle chamber to install a new gun nozzle, then moves to a probe reagent chamber to absorb a probe reagent and moves to a reaction chamber, then the probe reagent is dripped onto a cell chip in the reaction chamber, then the pipetting gun moves to a waste material treatment chamber, and the gun nozzle automatically falls off to the waste material treatment chamber;
(6) The liquid transferring gun moves to a gun nozzle chamber to install a new gun nozzle, then moves to a dye reagent chamber to absorb a coloring agent and moves to a reaction chamber, then the coloring agent is dripped on a cell chip in the reaction chamber, then the liquid transferring gun moves to a waste material treatment chamber, and the gun nozzle automatically falls off to the waste material treatment chamber;
(7) The temperature in the reaction chamber is raised to a certain temperature by a heating module and then kept for a period of time, and then the temperature is lowered and then kept for a period of time to carry out dyeing and hybridization reaction.
Preferably, after the staining and hybridization reaction, moving the pipetting gun to a gun nozzle chamber to install a new gun nozzle, then moving the pipetting gun to a cleaning chamber to absorb the cleaning solution and moving the pipetting gun to the reaction chamber, wherein the pipetting gun firstly drips one part of the cleaning solution on the cell chip, and drips the other part of the cleaning solution after a period of time; then the pipette is moved to the waste treatment chamber, the gun nozzle automatically falls off to the waste treatment chamber, then the pipette is moved back to the reaction chamber, and air is blown out through the pipette to blow dry the cell chips; then the liquid transferring gun moves to a gun nozzle chamber to install a new gun nozzle, then moves to a display liquid chamber to absorb display liquid and moves to a reaction chamber, the display liquid is dripped onto a cell chip in the reaction chamber, then the liquid transferring gun moves to a waste material treatment chamber, and the gun nozzle automatically falls off to the waste material treatment chamber; finally, taking out the cell chip in the reaction chamber, sealing the cell chip, and then scanning and image analysis are carried out through a scanner.
According to the technical scheme, the sample chamber, the consumable chamber, the reaction chamber, the waste treatment and the like are arranged on the detection frame, and reagents are automatically dripped into a plurality of cell chips in the reaction chamber through the pipette to perform dyeing and hybridization reactions, so that the processes of dyeing, hybridization, slide pushing and the like are integrated, the full-automatic detection of the circulating tumor cells in the whole process can be formed by matching with a centrifugal process, the detection efficiency is greatly improved, the circulating tumor cells can be accurately and effectively captured and identified through a chromosome probe/immune dyeing identification method, the false negative is reduced, and the detection sensitivity is greatly improved.
Drawings
FIG. 1 is a schematic top view of the detecting device of the present invention.
FIG. 2 is a schematic diagram of a forward structure of the detecting device of the present invention.
Detailed Description
The invention will now be described in detail with reference to fig. 1 and 2, wherein the exemplary embodiments and descriptions of the invention are provided to explain the invention, but not to limit the invention.
As shown in fig. 1 and 2, the present invention provides a circulating tumor cell detection apparatus, which includes a detection frame 1 and a plurality of sample test tubes 2 centrifuged by a centrifuge, wherein each sample test tube is filled with serum containing tumor cells, the detection frame is provided with a sample chamber 3, a consumable chamber 4 and a reaction chamber 5, the plurality of sample test tubes are placed in the sample chamber, a plurality of cell chips 6 are placed in the reaction chamber, the serum in each sample test tube is injected onto the corresponding cell chip through a pipetting gun 7 arranged on the detection frame, and then a plurality of reagents in the consumable chamber are sequentially dripped onto the corresponding cell chip through the pipetting gun to perform staining and hybridization reactions on the tumor cells on the cell chip, so as to accurately and effectively capture and identify the circulating tumor cells, and reduce false negatives. In the implementation process, a gun nozzle chamber 41, a reagent consumable chamber and the like are arranged in the consumable chamber 4, a plurality of gun nozzles 71 matched with the liquid transfer gun are placed in the gun nozzle chamber, and the liquid transfer gun is moved to the gun nozzle chamber before liquid suction at each time to automatically replace new gun nozzles, so that the cleanness and sanitation of a dripping reagent are guaranteed, cross contamination is avoided, and the detection precision is improved; and moreover, automation is realized, and the detection efficiency is improved. The detection frame is also provided with a waste material treatment chamber 8, and the gun nozzle of the liquid-transferring gun after liquid transfer is finished automatically falls off to the waste material treatment chamber, so that the used gun nozzle can be collected and treated from the waste material treatment chamber in a centralized manner, and the sanitary environment of the detection equipment is further improved.
Specifically, the consumable chamber 4 such as a reagent consumable chamber is provided with a fixed liquid chamber 42, a washing liquid chamber 43, an ethanol chamber 44, a probe reagent chamber 45, a dye reagent chamber 46, a display liquid chamber 47, and the like, and various reagent bottles can be used as the fixed liquid chamber, the washing liquid chamber, the ethanol chamber, the probe reagent chamber, the dye reagent chamber, and the display liquid chamber. After serum in the sample test tube is injected into the cell chip, the pipette sequentially drips the fixing liquid in the fixing liquid chamber, the cleaning liquid in the cleaning liquid chamber, the ethanol in the ethanol chamber, the probe reagent in the probe reagent chamber and the dye reagent in the dye reagent chamber on the cell chip for dyeing and hybridization reaction; after the reaction is finished, the cleaning liquid in the cleaning liquid chamber and the display liquid in the display liquid chamber are dripped on the cell chip in sequence through the pipette to be cleaned and displayed, so that scanning and image analysis can be performed through a scanner. The invention integrates the procedures of dyeing, hybridization, slide pushing and the like, can be matched with a centrifugal procedure to form full-flow circulating tumor cell full-automatic detection, and greatly improves the detection efficiency; and the whole detection equipment has the advantages of compact structure, small volume, simple operation and low cost.
Preferably, the detection frame is provided with a guide rail 72 positioned above the sample chamber, the fixed liquid chamber, the cleaning liquid chamber, the ethanol chamber, the probe reagent chamber, the dye reagent chamber, the display liquid chamber, the gun nozzle chamber, the reaction chamber and the waste treatment chamber, the guide rail is provided with an automatic guide trolley 73, and the automatic guide trolley drives the liquid-transferring gun mounted on the automatic guide trolley to move along the guide rail. The automatic guided vehicle is internally provided with a control program, and the automatic guided vehicle is enabled to complete corresponding operation according to a specified path and time through program design. The liquid transferring gun comprises a gun body and a gun nozzle, the gun body drives the gun nozzle to automatically suck and drip reagent through automatic expansion according to a software program, and the automatic falling and mounting process of the gun nozzle adopts the prior art; and the gun body is connected with an air source, and dry air can be automatically and timely sprayed according to a designed program.
The reaction chamber is a constant-temperature closed chamber, an automatic chamber door 51 is arranged on the constant-temperature closed chamber, when the automatic guide trolley moves to the position of the automatic chamber door, the automatic chamber door is automatically opened, the automatic guide trolley drives the liquid transfer gun to enter the constant-temperature closed chamber along the guide rail, and the liquid transfer gun drips the corresponding reagent on the corresponding cell chip in the constant-temperature closed chamber, so that the reagent is dripped automatically by the liquid transfer gun, the reaction chamber can be closed by the automatic chamber door, the rapid heating and heat preservation are realized, and the hybridization and dyeing effects are improved. Preferably, the constant-temperature closed bin is divided into an upper chamber and a lower chamber by a bedplate 52, a liquid injection platform 54 is arranged in the upper chamber 53 at the upper side of the bedplate, and a plurality of cell chips can be placed on the liquid injection platform at the same time so as to improve the efficiency; a heating module 56 which can heat and insulate the upper chamber is arranged in the lower chamber 55 at the lower side of the bedplate, wherein the heating module can adopt a temperature controller and a heating wire which are arranged at the lower side of the bedplate and a temperature sensor which is arranged at the upper chamber, thereby realizing the accurate control of the temperature in the reaction chamber.
The sample chamber is internally provided with the test tube rack 31, and the plurality of sample test tubes are inserted on the test tube rack, so that the sample chamber is more favorable for placing a plurality of sample test tubes, can ensure that the sample test tubes are more stable, and is convenient for the pipette to absorb sample serum. Preferably, the waste material treatment chamber is a closed space surrounded by a plurality of partition plates 81, an opening 82 is formed in the upper partition plate, and the fallen gun nozzle enters the closed space through the opening, so that the used gun nozzle is conveniently collected. The invention realizes the full-automatic detection of the tumor cells by matching the automatic guide trolley and the pipette with a plurality of chambers, thereby greatly improving the detection efficiency.
The invention also provides a using method of the circulating tumor cell detection equipment, wherein 8 or 12 chips and the like can be placed in the reaction chamber, 3mL of stationary liquid (3 mL of 50 muL X5 reaction cavities/test piece X12 test pieces) in the stationary liquid chamber, 12mL of cleaning liquid, 6mL of ethanol, 600 muL of probe reagent, 60 muL of coloring agent, 600 muL of display liquid and 600-660 gun nozzles are adopted, and the method specifically comprises the following steps: firstly, placing a plurality of sample test tubes after centrifugal separation on a test tube rack of a sample chamber, moving a liquid transfer gun to a gun nozzle chamber to automatically install a gun nozzle, then moving the liquid transfer gun to the sample chamber to absorb 50 mu L of serum in the sample test tubes and moving the serum to a reaction chamber, then injecting the serum onto a cell chip in the reaction chamber, then moving the liquid transfer gun to a waste material treatment chamber, and automatically dropping the gun nozzle into the waste material treatment chamber. The liquid transfer gun moves to a gun nozzle chamber to install a new gun nozzle, then moves to a stationary liquid chamber to absorb 50 mu L of stationary liquid, moves to a reaction chamber, then drips the stationary liquid onto a cell chip in the reaction chamber, then moves to a waste treatment chamber, and the gun nozzle automatically falls off to the waste treatment chamber; then the pipette is moved back to the reaction chamber, after the cell chip is incubated for 10min, air is blown out by the pipette for 10min to blow and dry the cell chip.
Then, the pipetting gun moves to a gun nozzle chamber to install a new gun nozzle, then moves to a cleaning chamber to absorb 100 mu L of cleaning solution and moves to a reaction chamber, the pipetting gun firstly drips 50 mu L of cleaning solution to the cell chip, and 50 mu L of cleaning solution is dripped after 5 min; then the liquid transferring gun is moved to a waste treatment chamber, and the gun nozzle automatically falls off into the waste treatment chamber; moving a liquid transfer gun to a gun nozzle chamber to install a new gun nozzle, then moving the liquid transfer gun to an ethanol chamber to absorb 100 mu L of ethanol and moving the liquid transfer gun to a reaction chamber, firstly dripping 50 mu L of ethanol on the cell chip, and dripping 50 mu L of ethanol after 5 min; and then moving the liquid transfer gun to the waste treatment chamber, automatically dropping the gun nozzle to the waste treatment chamber, moving the liquid transfer gun back to the reaction chamber, and blowing gas through the liquid transfer gun to blow dry the cell chip after the cell chip is incubated for 2 min.
The liquid transferring gun moves to a gun nozzle chamber to install a new gun nozzle, then moves to a probe reagent chamber to absorb 10 mu L of probe reagent and moves to a reaction chamber, then the probe reagent is dripped on a cell chip in the reaction chamber, then the liquid transferring gun moves to a waste material treatment chamber, and the gun nozzle automatically falls off to the waste material treatment chamber; then the liquid transferring gun moves to a gun nozzle chamber to install a new gun nozzle, then moves to a dye reagent chamber to absorb 1 mu L of coloring agent and moves to a reaction chamber, then the coloring agent is dripped on a cell chip in the reaction chamber, then the liquid transferring gun moves to a waste material treatment chamber, and the gun nozzle automatically falls off to the waste material treatment chamber; and then heating the reaction chamber to 85 ℃ through a heating module and keeping the temperature for 10min, and then cooling the reaction chamber to 37 ℃ and keeping the temperature for 2h so as to perform dyeing and hybridization reaction.
After the dyeing and hybridization reaction is finished, moving a liquid transfer gun to a gun nozzle chamber to install a new gun nozzle, then moving the liquid transfer gun to a cleaning chamber to absorb 100 mu L of cleaning solution and moving the liquid transfer gun to the reaction chamber, wherein the liquid transfer gun firstly drips 50 mu L of cleaning solution on the cell chip, and later drips 50 mu L of cleaning solution; then moving the liquid transfer gun to a waste treatment chamber, automatically dropping a gun nozzle into the waste treatment chamber, moving the liquid transfer gun back to the reaction chamber, and blowing air out for 10min through the liquid transfer gun to blow dry the cell chip; the liquid transfer gun moves to a gun nozzle chamber to install a new gun nozzle, then moves to a display liquid chamber to absorb 10 mu L of display liquid and moves to a reaction chamber, then the display liquid is dripped on a cell chip in the reaction chamber, then the liquid transfer gun moves to a waste material treatment chamber, and the gun nozzle automatically falls off to the waste material treatment chamber; finally, taking out the cell chip in the reaction chamber, sealing the cell chip, and then scanning and analyzing images through a scanner, thereby realizing the detection of the circulating tumor cells of the solid cancer.
Therefore, the invention adopts the technologies of synchronous immunofluorescence staining, in-situ hybridization and the like, greatly improves the capture rate of circulating tumor cell detection and the cell identification accuracy rate, and greatly reduces false negative. The disposable gun nozzle can extract a leucocyte layer, add magnetic beads, arrange an inclination angle, shake the blending module for automatic blending, move to the strong magnetic module, extract liquid without the magnetic beads from the disposable gun nozzle, clean cells, and transmit and transfer antibodies, probe reagents and related washing liquid from the disposable gun nozzle, so that the steps of dyeing and hybridization are completed; and (4) mounting the slide, moving the slide to an automatic scanner, scanning the slide, and carrying out artificial intelligent analysis on the image to identify CTC. The sample flux can reach 48 cases every day, and the detection efficiency is greatly improved.
Claims (10)
1. The utility model provides a circulation tumor cell check out test set, includes test rack and by the several sample test tube after centrifuge centrifugation, each sample test tube is equipped with the serum that contains tumor cell in, its characterized in that: the detection frame is provided with a sample chamber, a consumable chamber and a reaction chamber, a plurality of sample test tubes are placed in the sample chamber, a plurality of cell chips are placed in the reaction chamber, serum in each sample test tube is injected onto the corresponding cell chip through a liquid transfer gun arranged on the detection frame, and then a plurality of reagents in the consumable chamber are dripped onto the corresponding cell chip in sequence through the liquid transfer gun so as to dye and perform hybridization reaction on tumor cells on the cell chip; the indoor stationary liquid room that is equipped with of consumptive material, washing liquid room, ethanol room, probe reagent room, dye reagent room and demonstration liquid room, after the cell chip was injected into to the serum, the pipetting gun is dripped indoor stationary liquid, the indoor washing liquid of washing liquid, the indoor ethanol of ethanol, the indoor probe reagent of probe reagent and the indoor dye reagent of stationary liquid in proper order on this cell chip in order to carry out dyeing and hybridization reaction, and the reaction is accomplished the back, the rethread the pipetting gun is dripped indoor washing liquid of washing liquid and the indoor demonstration liquid of demonstration liquid in proper order on the cell chip in order to wash and show.
2. The circulating tumor cell detecting apparatus according to claim 1, wherein: the liquid-transferring gun is characterized in that a gun nozzle chamber is arranged in the consumable chamber, a plurality of gun nozzles matched with the liquid-transferring gun are placed in the gun nozzle chamber, and the liquid-transferring gun moves to the gun nozzle chamber before liquid suction at each time and is automatically replaced by a new gun nozzle.
3. The circulating tumor cell detecting apparatus according to claim 2, wherein: the detection frame is provided with a waste treatment chamber, and a gun nozzle of the liquid transfer gun automatically falls to the waste treatment chamber after liquid transfer is completed.
4. The circulating tumor cell detecting apparatus according to claim 3, wherein: the waste material treatment chamber is a closed space surrounded by a plurality of partition plates, an opening is formed in the partition plate on the upper side, and the fallen gun nozzle enters the closed space through the opening.
5. The circulating tumor cell detecting apparatus according to claim 1, 2, 3 or 4, wherein: the detection frame is provided with a guide rail which is positioned above the sample chamber, the fixed liquid chamber, the cleaning liquid chamber, the ethanol chamber, the probe reagent chamber, the dye reagent chamber, the display liquid chamber, the gun nozzle chamber, the reaction chamber and the waste treatment chamber, the guide rail is provided with an automatic guide trolley, and the automatic guide trolley drives the liquid-transferring gun which is arranged on the automatic guide trolley to move along the guide rail.
6. The circulating tumor cell detecting apparatus according to claim 5, wherein: the reaction chamber is a constant-temperature closed bin, an automatic bin gate is arranged on the constant-temperature closed bin, when the automatic guide trolley moves to the position of the automatic bin gate, the automatic bin gate is automatically opened, the automatic guide trolley drives the liquid-transferring gun to enter the constant-temperature closed bin along the guide rail, and the liquid-transferring gun drips a corresponding reagent on a corresponding cell chip in the constant-temperature closed bin.
7. The circulating tumor cell detecting apparatus according to claim 6, wherein: the constant-temperature closed bin is divided into an upper chamber and a lower chamber by a bedplate, the upper side of the bedplate is positioned in the upper chamber and is provided with a liquid injection platform, a plurality of cell chips are arranged on the liquid injection platform, and the lower side of the bedplate is positioned in the lower chamber and is provided with a heating module which can heat and insulate the upper chamber.
8. The circulating tumor cell detecting apparatus according to claim 1, wherein: a test tube rack is arranged in the sample chamber, and a plurality of sample test tubes are inserted on the test tube rack.
9. A method of using the circulating tumor cell detecting apparatus of any one of claims 1 to 8, comprising the steps of:
(1) Placing the centrifugally separated sample test tube in a sample chamber, moving a liquid transfer gun to a gun nozzle chamber to automatically install a gun nozzle, then moving the sample chamber to absorb serum in the sample test tube and moving the serum to a reaction chamber, then injecting the serum onto a cell chip in the reaction chamber, then moving the liquid transfer gun to a waste treatment chamber, and automatically dropping the gun nozzle into the waste treatment chamber;
(2) The liquid transferring gun moves to a gun nozzle chamber to install a new gun nozzle, then moves to a stationary liquid chamber to absorb stationary liquid and moves to a reaction chamber, the stationary liquid is dripped onto a cell chip in the reaction chamber, then the liquid transferring gun moves to a waste treatment chamber, and the gun nozzle automatically falls off to the waste treatment chamber; then the pipetting gun moves back to the reaction chamber, after the cell chip is incubated for a period of time, air is blown out through the pipetting gun to blow dry the cell chip;
(3) Moving a liquid transfer gun to a gun nozzle chamber to install a new gun nozzle, then moving the liquid transfer gun to a cleaning chamber to absorb cleaning liquid and moving the cleaning liquid to a reaction chamber, wherein the liquid transfer gun firstly drips one part of the cleaning liquid on the cell chip, and drips the other part of the cleaning liquid after a period of time; then the liquid transferring gun is moved to a waste treatment chamber, and the gun nozzle automatically falls off into the waste treatment chamber;
(4) Moving a liquid transfer gun to a gun nozzle chamber to install a new gun nozzle, then moving the liquid transfer gun to an ethanol chamber to absorb ethanol and moving the liquid transfer gun to a reaction chamber, dripping one part of ethanol to the cell chip by the liquid transfer gun, and dripping the other part of ethanol after a period of time; moving the liquid transfer gun to a waste treatment chamber, automatically dropping a gun nozzle to the waste treatment chamber, moving the liquid transfer gun back to the reaction chamber, and blowing out gas through the liquid transfer gun to blow dry the cell chip after the cell chip is incubated for a period of time;
(5) The liquid transferring gun moves to a gun nozzle chamber to install a new gun nozzle, then moves to a probe reagent chamber to absorb a probe reagent and moves to a reaction chamber, the probe reagent is dripped onto a cell chip in the reaction chamber, then the liquid transferring gun moves to a waste material processing chamber, and the gun nozzle automatically falls off to the waste material processing chamber;
(6) The liquid transferring gun moves to a gun nozzle chamber to install a new gun nozzle, then moves to a dye reagent chamber to absorb a coloring agent and moves to a reaction chamber, then the coloring agent is dripped on a cell chip in the reaction chamber, then the liquid transferring gun moves to a waste material treatment chamber, and the gun nozzle automatically falls off to the waste material treatment chamber;
(7) The temperature in the reaction chamber is raised to a certain temperature by a heating module and then kept for a period of time, and then the temperature is lowered and kept for a period of time to carry out dyeing and hybridization reactions.
10. The method of using the circulating tumor cell detecting apparatus according to claim 9, wherein: after the dyeing and hybridization reaction is finished, moving a liquid transfer gun to a gun nozzle chamber to install a new gun nozzle, then moving the liquid transfer gun to a cleaning chamber to absorb cleaning liquid and moving the liquid transfer gun to the reaction chamber, wherein the liquid transfer gun firstly drips one part of the cleaning liquid on the cell chip, and drips the other part of the cleaning liquid after a period of time; then the pipette is moved to a waste treatment chamber, the nozzle of the pipette automatically falls off into the waste treatment chamber, then the pipette is moved back to the reaction chamber, and air is blown out through the pipette to blow dry the cell chip; then the liquid transferring gun moves to a gun nozzle chamber to install a new gun nozzle, then moves to a display liquid chamber to absorb display liquid and moves to a reaction chamber, the display liquid is dripped onto a cell chip in the reaction chamber, then the liquid transferring gun moves to a waste material treatment chamber, and the gun nozzle automatically falls off to the waste material treatment chamber; finally, taking out the cell chip in the reaction chamber, sealing the cell chip, and then scanning and image analyzing the cell chip by a scanner.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211120006.3A CN115201476A (en) | 2022-09-15 | 2022-09-15 | Circulating tumor cell detection equipment and use method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211120006.3A CN115201476A (en) | 2022-09-15 | 2022-09-15 | Circulating tumor cell detection equipment and use method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115201476A true CN115201476A (en) | 2022-10-18 |
Family
ID=83572437
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211120006.3A Pending CN115201476A (en) | 2022-09-15 | 2022-09-15 | Circulating tumor cell detection equipment and use method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115201476A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2024082187A1 (en) * | 2022-10-19 | 2024-04-25 | 深圳华大生命科学研究院 | Pipetting apparatus for fluidic chip, and pipetting method for fluidic chip |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104062162A (en) * | 2014-07-10 | 2014-09-24 | 上海交通大学医学院附属瑞金医院 | Three-dimensional nanometer chip, method for fractionation detection on circulating tumor cells (CTC) by using chip, and application of three-dimensional nanometer chip |
| CN105785005A (en) * | 2016-04-13 | 2016-07-20 | 杭州华得森生物技术有限公司 | Circulating tumor cell detection kit and application thereof |
| CN106596941A (en) * | 2016-12-21 | 2017-04-26 | 上海市质子重离子医院有限公司 | Detection method for detecting prostate cancer circulating tumor cells |
| CN206298600U (en) * | 2016-12-23 | 2017-07-04 | 西安医臻生物医药科技有限公司 | A kind of circulating tumor cell capture agent box |
| CN106970224A (en) * | 2017-03-16 | 2017-07-21 | 武汉康录生物技术股份有限公司 | A kind of kit of application CD45 immunofluorescences joint CEP probe identification circulating tumor cells and its application |
| CN108441477A (en) * | 2017-05-22 | 2018-08-24 | 浙江大学 | A method of capture cancer of pancreas circulating tumor cell is enriched with based on CSV antibody |
| CN110940568A (en) * | 2019-12-17 | 2020-03-31 | 武汉友芝友医疗科技股份有限公司 | Integrated system for cell enrichment, separation, dyeing and flaking |
| CN112391263A (en) * | 2020-11-18 | 2021-02-23 | 深圳市儿童医院 | Neuroblastoma circulating tumor cell capturing chip and manufacturing method thereof |
| CN113088433A (en) * | 2021-04-09 | 2021-07-09 | 天津市第三中心医院 | Full-automatic cell processing pulsating workstation |
| CN114487415A (en) * | 2021-12-24 | 2022-05-13 | 江苏莱尔生物医药科技有限公司 | Tumor cell PD-L1PD-L2 immunofluorescence + FISH detection kit |
-
2022
- 2022-09-15 CN CN202211120006.3A patent/CN115201476A/en active Pending
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104062162A (en) * | 2014-07-10 | 2014-09-24 | 上海交通大学医学院附属瑞金医院 | Three-dimensional nanometer chip, method for fractionation detection on circulating tumor cells (CTC) by using chip, and application of three-dimensional nanometer chip |
| CN105785005A (en) * | 2016-04-13 | 2016-07-20 | 杭州华得森生物技术有限公司 | Circulating tumor cell detection kit and application thereof |
| CN106596941A (en) * | 2016-12-21 | 2017-04-26 | 上海市质子重离子医院有限公司 | Detection method for detecting prostate cancer circulating tumor cells |
| CN206298600U (en) * | 2016-12-23 | 2017-07-04 | 西安医臻生物医药科技有限公司 | A kind of circulating tumor cell capture agent box |
| CN106970224A (en) * | 2017-03-16 | 2017-07-21 | 武汉康录生物技术股份有限公司 | A kind of kit of application CD45 immunofluorescences joint CEP probe identification circulating tumor cells and its application |
| CN108441477A (en) * | 2017-05-22 | 2018-08-24 | 浙江大学 | A method of capture cancer of pancreas circulating tumor cell is enriched with based on CSV antibody |
| CN110940568A (en) * | 2019-12-17 | 2020-03-31 | 武汉友芝友医疗科技股份有限公司 | Integrated system for cell enrichment, separation, dyeing and flaking |
| CN112391263A (en) * | 2020-11-18 | 2021-02-23 | 深圳市儿童医院 | Neuroblastoma circulating tumor cell capturing chip and manufacturing method thereof |
| CN113088433A (en) * | 2021-04-09 | 2021-07-09 | 天津市第三中心医院 | Full-automatic cell processing pulsating workstation |
| CN114487415A (en) * | 2021-12-24 | 2022-05-13 | 江苏莱尔生物医药科技有限公司 | Tumor cell PD-L1PD-L2 immunofluorescence + FISH detection kit |
Non-Patent Citations (2)
| Title |
|---|
| 张金玲 等: "基于微流控芯片的免疫反应快速检测系统研究", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
| 陈超 等: "《新技术与精准医学》", 31 December 2017 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2024082187A1 (en) * | 2022-10-19 | 2024-04-25 | 深圳华大生命科学研究院 | Pipetting apparatus for fluidic chip, and pipetting method for fluidic chip |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104745452B (en) | Rare cell automates capture device | |
| CN107656085B (en) | Blood detector | |
| WO2018126775A1 (en) | Automatic analysis device and sample analysis method | |
| CN101581721B (en) | Have at least one provide in centrifugation cycle in advance cohesion assessment imager immunologic diagnosises testing equipment | |
| CN108048315A (en) | It is a kind of based on the fluorescence quantitative PCR instrument being automatically loaded | |
| CN115201476A (en) | Circulating tumor cell detection equipment and use method thereof | |
| CN106226540B (en) | Full-automatic protein chip analyzer | |
| JP2023546265A (en) | Fully automatic detached cell specimen preparation method | |
| JP2023510189A (en) | Automated staining system and reaction chamber | |
| CN111638357A (en) | Immunofluorescence kit and method for E-Cadherin mutation of peripheral blood circulating tumor cells of patient with non-small cell lung cancer | |
| CN111521794A (en) | Immunofluorescence kit and detection method for detecting NSE gene mutation of peripheral blood circulating tumor cells of small cell lung cancer patients | |
| CN111562375A (en) | Immunofluorescence kit for detecting expression of peripheral blood circulating tumor cells PD-L1 of gastric cancer patient and detection method | |
| CN114487415A (en) | Tumor cell PD-L1PD-L2 immunofluorescence + FISH detection kit | |
| CN112433047A (en) | Quantitative excrement detection analyzer | |
| US11719712B2 (en) | Automated sample deposition and staining systems and associated methods | |
| CN111521799A (en) | Immunofluorescence kit for detecting esophageal squamous carcinoma patient PD-L1 gene expression through peripheral blood circulation tumor cells | |
| CN111551718A (en) | Immunofluorescence kit and detection method for detecting prostate cancer patient peripheral blood circulating tumor cell PD-L1 expression | |
| CN111521793A (en) | Immunofluorescence kit and detection method for detecting CEA gene mutation of peripheral blood circulating tumor cells of non-small cell lung cancer patients | |
| CN206258461U (en) | A kind of reaction cup automatic flushing device on time resolution detector | |
| EP3160648A1 (en) | Specimen processing systems, pipette assemblies and methods for preparing reagents | |
| TWM619821U (en) | Automatic processing device for liquid samples | |
| CN210294289U (en) | Full-automatic biochemical analyzer | |
| WO2019127959A1 (en) | Rapid detection system and method involving biochemical detection, enzyme immunoassay and chemiluminescence detection | |
| CN111521800A (en) | Immunofluorescence kit for detecting gene expression of PD-L1 of colorectal cancer patient through peripheral blood circulating tumor cells | |
| CN111551727A (en) | Immunofluorescence kit and detection method for detecting ALK gene mutation of peripheral blood circulating tumor cells of non-small cell lung cancer patients |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20221018 |