CN108693115A - NK cell detection methods and system - Google Patents
NK cell detection methods and system Download PDFInfo
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- CN108693115A CN108693115A CN201711416647.2A CN201711416647A CN108693115A CN 108693115 A CN108693115 A CN 108693115A CN 201711416647 A CN201711416647 A CN 201711416647A CN 108693115 A CN108693115 A CN 108693115A
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- 210000000822 natural killer cell Anatomy 0.000 title claims abstract description 86
- 238000001514 detection method Methods 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 claims abstract description 16
- 230000005540 biological transmission Effects 0.000 claims abstract description 10
- 238000003708 edge detection Methods 0.000 claims abstract description 10
- 210000004698 lymphocyte Anatomy 0.000 claims abstract description 7
- 238000004458 analytical method Methods 0.000 claims description 27
- 238000012937 correction Methods 0.000 claims description 7
- 238000003709 image segmentation Methods 0.000 claims description 6
- 230000017531 blood circulation Effects 0.000 claims description 3
- 230000001737 promoting effect Effects 0.000 claims description 3
- 238000001000 micrograph Methods 0.000 claims 1
- 238000012543 microbiological analysis Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 11
- 206010028980 Neoplasm Diseases 0.000 description 6
- 201000011510 cancer Diseases 0.000 description 5
- 230000011218 segmentation Effects 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 101100117236 Drosophila melanogaster speck gene Proteins 0.000 description 2
- 238000004026 adhesive bonding Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000002651 drug therapy Methods 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 238000003703 image analysis method Methods 0.000 description 2
- 230000002147 killing effect Effects 0.000 description 2
- 244000045947 parasite Species 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010041591 Spinal osteoarthritis Diseases 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000001772 blood platelet Anatomy 0.000 description 1
- 230000022534 cell killing Effects 0.000 description 1
- 230000002490 cerebral effect Effects 0.000 description 1
- 208000036319 cervical spondylosis Diseases 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 208000030172 endocrine system disease Diseases 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000005089 fruit drop Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 229940088592 immunologic factor Drugs 0.000 description 1
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- 238000009434 installation Methods 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 210000002751 lymph Anatomy 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
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- 230000003287 optical effect Effects 0.000 description 1
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
- G01N21/85—Investigating moving fluids or granular solids
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
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- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention provides a kind of NK cell detection methods and systems, are related to biology, microbiological analysis detection field, and method includes:It obtains comprising the micro-image including NK cells;The micro-image of leucocyte is obtained again, it can be seen that contains light transmission spot in leucocyte;Edge detection is carried out, and fills edge internal voids, to form complete NK cell shapes;Calculate the difference in areas of the front and back NK cells of filling;When higher than the first setting value, then it is judged as lymphocyte;The image in NK cell edges is split again, calculates the area and circularities of leucocyte iuuminting spot, when higher than the second setting value, then can determine that as NK cells.The complicated processes such as not only time of measuring is short, need not dye culture are easy.
Description
Technical field
The present invention relates to biology, microbiological analysis detection fields, more particularly, to a kind of NK cell detection methods and system.
Background technology
The development of modern medicine is largely dependent upon laboratory facilities and instrument and equipment, wherein microscopical application is very
Extensively, by photo-translating system, and the image under microscope can be out shown and is analyzed by computer acquisition.
Simple microscope can do many analysis and experiment, but in order to expand the observation scope of microcosmos, develop again later
Go out super-high power microscopic analysis device, effectively increase amplification factor, observes apparent intuitive to biological cell and microorganism, see
More details are observed, the accuracy of analysis is improved.In micro image collection to computer, it can be shown, be deposited
Shelves and record printing, and can also achieve the purpose that further to analyze image using the analysis means of computer, it obtains
Better analytical effect.Objective table that objective table is moved automatically using tri- axis of XYZ, it can be achieved that automatic focusing function of walking on stage automatically,
The degree of automation higher.
The killing vigor of NK cells is big, and lethal effect occurs early after acting on target cell, and the target cell of NK cells mainly has
Certain tumour cells (including part cell line), virus infected cell, certain autologous tissue's cells (such as haemocyte), parasite
Deng, therefore NK cells are that body is antitumor, anti-infectious important immune factor, are combined with immunocyte detection, and carry out
The important detection methods of cancer early warning.
Currently, the method for carrying out NK cell detections in clinic is seldom, mainly there is flow cytometry (FCM), it is to pass through spy
Fixed fluorescer is dyed, and is measured to measure to fluoresced luminosity, measures complicated, long time period, noise
More relatively low than the high precision that causes, precision is very poor when especially NK cell contents especially lack, and is not suitable for the early stage hair of NK cells
It is existing.The method for directly carrying out NK cell image detections is to utilize Bu Shi microscopes, and finger tip 1 is only needed to bleed, so that it may artificial to carry out
It has detected.Certain Bu Shi microscopes can also be analyzed and be assessed to everyone body present situation, disease risk and future trend,
Diagnostic image most direct, clear, timely and accurately and opinion are provided to diagnosis person, it is thin in blood from seeing on color/graphics
The variations such as solid shape, structure and the quantity of born of the same parents, blood platelet, bacterium, fungi and parasite etc., by observing vivo oxidation certainly
By the case where base and trace existing for free radical, tumour, inflammation, Cardial or cerebral vascular diseases, endocrine system disease, mistake are judged with this
Quick and cervical spondylosis etc. in a few minutes can do the physical condition of human body one accurate, comprehensive assessment.
It is checked by microscopic system, is all judged by artificialvision's observation at present, this is because figure
As automatic identification technology it is also unqualified, cannot automatic decision, can only manually judge.The numbers of NK cells is also manually to see
It examines, as qualitative analysis, is also possible, but met the requirements for quantitative analysis, accuracy or hardly possible.NK cell killings
Power is huge, and being human body has great virus infection or the killing immunocyte that just can accordingly occur after cancer cell occur, no matter from
The angle resisted the disease, the especially angle of carly fruit drop cancer, the accurate NK cells that measure all are in demand, this is also one
A hard nut to crack in currently available technology, cannot also accomplish the method measured by MIcrosope image come automatic measurement NK
Practical degree is also not achieved in cell, precision, can only indirect staining measure, precision is low, process is complicated.
Invention content
The purpose of the present invention is to provide a kind of NK cell detection methods and systems, to solve manually seeing in the prior art
In terms of examining NK cells, existing technical difficulty is big, accuracy of detection is low, process is complicated.
The present invention provides a kind of NK cell detection methods, including:
It obtains comprising the micro-image including NK cells;
The micro-image of leucocyte is obtained again, it can be seen that contains light transmission spot in leucocyte;
Edge detection is carried out, and fills edge internal voids, to form complete NK cell shapes;
Calculate the difference in areas of the front and back NK cells of filling;When higher than the first setting value, then it is judged as lymphocyte;
The image in NK cell edges is split again, calculates the area and circularities of leucocyte iuuminting spot,
When higher than the second setting value, then it can determine that as NK cells.
In the above-mentioned technical solutions, further, it obtains comprising the micro-image including NK cells:
Microscopic system obtains human body promoting blood circulation image;
Image is acquired by the video heads being fixed on microscopic system, after being converted into electric signal, is transmitted to analysis module.
In the above-mentioned technical solutions, further, in leucocyte containing the image in light transmission spot and NK cell edges all
It is that image segmentation is carried out by the method for automatic correction threshold, improves the accuracy of edge detection.
In addition, the application also provides a kind of NK cell detecting systems, including:
Microscopic system, the microscopic system be used to obtain comprising including NK cells micro-image and obtain again it is white carefully
The micro-image of born of the same parents;
Image information collecting module, described image information acquisition module are connect with microscopic system, for receiving and locating
Manage micro-image;
Analysis module, the analysis module are connect with image information collecting module and control module, for analyzing acquisition
Micro-image;Edge detection is carried out, and fills edge internal voids, to form complete NK cell shapes;It calculates before and after filling
The difference in areas of NK cells;When higher than the first setting value, then it is judged as lymphocyte;The image in NK cell edges is carried out again
Segmentation, calculate the area and circularities of leucocyte iuuminting spot, when higher than the second setting value, then can determine that for NK it is thin
Born of the same parents;
Output module, the output module are connect with analysis module, the analysis result for exporting analysis module.
In the above-mentioned technical solutions, further, the output module is printer.
In the above-mentioned technical solutions, further, the microscopic system further includes that can realize auto-translating and focusing
Three axis move objective table automatically.
In the above-mentioned technical solutions, further, the microscopic system includes that microscope and lens group are constituted, described
Microscope group is connect with microscope, and the video heads are fixed in lens group.
In the above-mentioned technical solutions, further, the lens group is formed by multiple lens gluings.
In the above-mentioned technical solutions, further, further include control module, the control module one side and analysis module
Connection, on the other hand connect with microscopic system.
In the above-mentioned technical solutions, further, described image information acquisition module is that digital video head or simulation regard
Frequency head.
Compared to the prior art, NK cell detection methods provided by the invention are adopted to the micro-image of NK cells
Sample carries out image using image analysis method, while to the image in NK cell edges by the method for automatic correction threshold
Segmentation, calculates the area and circularities of leucocyte iuuminting spot, when higher than setting value, then can determine that as NK cells, can
With the more accurate NK cells directly identified in human body, and that this time of measuring is short, need not dye culture etc. is complicated
Process, easy, reliable, for the early-stage cancer early warning and better effect and rehabilitation of observation drug therapy, provides one
Very effective detection mode.
The additional aspect and advantage of the present invention will become apparent in following description section, or practice through the invention
Recognize.
Description of the drawings
It, below will be to specific in order to illustrate more clearly of the specific embodiment of the invention or technical solution in the prior art
Embodiment or attached drawing needed to be used in the description of the prior art are briefly described, it should be apparent that, in being described below
Attached drawing is some embodiments of the present invention, for those of ordinary skill in the art, before not making the creative labor
It puts, other drawings may also be obtained based on these drawings.
Fig. 1 is the NK cell detection method flow diagrams that some embodiments of the present application provide;
Fig. 2 is the structural schematic diagram for the NK cell detecting systems that some embodiments of the present application provide;
Fig. 3 is the structural schematic diagram of the microscopic system for the NK cell detecting systems that some embodiments of the present application provide.
Reference numeral:
1- microscopic systems;2- image information collecting modules;3- analysis modules;4- output modules;5- control modules;11-
Microscope;12- lens groups;Tri- axis of 13- moves objective table automatically.
Specific implementation mode
Technical scheme of the present invention is clearly and completely described below in conjunction with attached drawing, it is clear that described implementation
Example is a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, ordinary skill
The every other embodiment that personnel are obtained without making creative work, shall fall within the protection scope of the present invention.
In the description of the present invention, it should be noted that term "center", "upper", "lower", "left", "right", "vertical",
The orientation or positional relationship of the instructions such as "horizontal", "inner", "outside" be based on the orientation or positional relationship shown in the drawings, merely to
Convenient for the description present invention and simplify description, do not indicate or imply the indicated device or element must have a particular orientation,
With specific azimuth configuration and operation, therefore it is not considered as limiting the invention.In addition, term " first ", " second ",
" third " is used for description purposes only, and is not understood to indicate or imply relative importance.
In the description of the present invention, it should be noted that unless otherwise clearly defined and limited, term " installation ", " phase
Even ", " connection " shall be understood in a broad sense, for example, it may be being fixedly connected, may be a detachable connection, or be integrally connected;It can
Can also be electrical connection to be mechanical connection;It can be directly connected, can also indirectly connected through an intermediary, Ke Yishi
Connection inside two elements.For the ordinary skill in the art, above-mentioned term can be understood at this with concrete condition
Concrete meaning in invention.
Refering to fig. 1, the present invention provides a kind of NK cell detection methods, including:
In step 110, it obtains comprising the micro-image including NK cells.
Wherein, the equipment for obtaining micro-image is microscopic system 1 and video heads, and microscopic system 1 includes microscope 11
It is constituted with lens group 12, the lens group 12 is connect with microscope 11, and the video heads are fixed in lens group 12.
Wherein, observation object is human body promoting blood circulation glass slide.
In the step 120, then the micro-image of leucocyte is obtained, it can be seen that contain light transmission spot in leucocyte.
The present embodiment is the segmentation of image to be carried out by the method for automatic correction threshold, and carry out side in image recognition
Edge detects, and is filled, finds out the size of filling.
In step 130, edge detection is carried out, and fills edge internal voids, to form complete NK cell shapes.
Because some are fuzzy for NK cell edges, if carrying out image segmentation with fixed threshold value, it is difficult to accurately detect side
Edge.Therefore, the present embodiment is the segmentation that image is carried out by the method for automatic correction threshold in image recognition, to ensure NK
The accuracy of cell edges detection.The method of the automatic correction threshold exactly carries out image segmentation, no with different threshold values
Same threshold value can obtain different NK cell area figures, select the threshold value wherein closest to average area as optimal threshold, can
To select fixed threshold, reliability is pursued.After selecting threshold value to complete edge detection, the gap of the inside is filled up, reforms into one
The shape of complete NK cells.
In step 140, the difference in areas of the front and back NK cells of filling is calculated;When higher than the first setting value, then it is judged as drenching
Bar cell.
It is split in step 150, then to the image in NK cell edges, calculates the face of leucocyte iuuminting spot
Product and circularities then can determine that when higher than the second setting value as NK cells.
Judge whether to be NK cells, one is lymphocyte to be judged whether, whether another is that have in cell to be judged
Than relatively round light transmission spot, it is determined that after NK cell edges, intramarginal image zooming-out is come out, then carry out different threshold values into
Row image segmentation, different threshold values can obtain different light transmission speck area figures, select wherein closest to the threshold value of average area
As optimal threshold, edge of the edge as light transmission spot after image segmentation calculates its area and circularity, thin if it is lymph
Born of the same parents, while light transmission speck area and circularity are more than setting value, then can determine that as NK cells.
Compared to the prior art, NK cell detection methods provided by the invention are adopted to the micro-image of NK cells
Sample carries out image using image analysis method, while to the image in NK cell edges by the method for automatic correction threshold
Segmentation, calculates the area and circularities of leucocyte iuuminting spot, when higher than setting value, then can determine that as NK cells, can
With the more accurate NK cells directly identified in human body, and that this time of measuring is short, need not dye culture etc. is complicated
Process, easy, reliable, for the early-stage cancer early warning and better effect and rehabilitation of observation drug therapy, provides one
Very effective detection mode.
Referring to Fig.2, the present invention provides a kind of NK cell detecting systems, including microscopic system 1, image information collecting module
2, analysis module 3 and output module 4.
Microscopic system 1, the microscopic system 1 be used to obtain comprising including NK cells micro-image and obtain thin in vain
The micro-image of born of the same parents;
Refering to Fig. 3, microscopic system 1 includes that microscope 11 and lens group 12 are constituted, the lens group 12 and microscope 11
Connection.
Image information collecting module 2, described image information acquisition module 2 are connect with microscopic system 1, for receive and
Handle micro-image;
In some embodiments, described image information acquisition module 2 is video heads, and video heads are fixed in lens group 12.
Specifically there are two types of modes for video heads:
One is optical signallings to become analog electrical signal by analog video head, is sent in collecting image of computer card, becomes
At the picture signal in computer;Another kind is to be mounted directly digital video head, no longer directly becomes number by image pick-up card
Word signal enters computer, such image definition higher, the distorted signals that do not convert.
Analysis module 3, the analysis module 3 are connect with image information collecting module 2 and control module 5, are obtained for analyzing
The micro-image taken;Edge detection is carried out, and fills edge internal voids, to form complete NK cell shapes;Calculate filling
The difference in areas of front and back NK cells;When higher than the first setting value, then it is judged as lymphocyte;Again to the image in NK cell edges
It is split, calculates the area and circularities of leucocyte iuuminting spot, when higher than the second setting value, then can determine that as NK
Cell;
Output module 4, the output module 4 are connect with analysis module 3, the analysis result for exporting analysis module 3.
In some embodiments, the output module 4 is printer, and printer prints out the result of analysis,
Including writings and image information.In some embodiments, the output module 4 is display screen, for showing analysis result.
In some embodiments, the microscopic system 1 further includes that can realize that three axis of auto-translating and focusing are automatic
Move objective table 13.
Objective table can be plain stage, can also be that three axis move objective table 13 automatically, objective table uses tri- axis of XYZ
Automatically the objective table moved is, it can be achieved that automatic focusing function of walking on stage automatically, the degree of automation higher.Specifically, XY axis controls
Stage surface moves, and automatic focusing function may be implemented in Z axis, and control Z axis variation acquires corresponding image, it is clear to analyze it
Degree, most clearly position is to focus optimum position.
In some embodiments, lens group 12 is formed by multiple lens gluings.
NK cell detecting systems further include control module 5, and 5 one side of the control module is connect with analysis module 3, another
Aspect is connect with microscopic system 1.Specifically, control module 5 controls three axis and moves the action of objective table 13 automatically, realizes automatic
Adjustment focusing.
Finally it should be noted that:The above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations;To the greatest extent
Present invention has been described in detail with reference to the aforementioned embodiments for pipe, it will be understood by those of ordinary skill in the art that:Its according to
So can with technical scheme described in the above embodiments is modified, either to which part or all technical features into
Row equivalent replacement;And these modifications or replacements, various embodiments of the present invention technology that it does not separate the essence of the corresponding technical solution
The range of scheme.
Claims (10)
1. a kind of NK cell detection methods, which is characterized in that including:
It obtains comprising the micro-image including NK cells;
The micro-image of leucocyte is obtained again, it can be seen that contains light transmission spot in leucocyte;
Edge detection is carried out, and fills edge internal voids, to form complete NK cell shapes;
Calculate the difference in areas of the front and back NK cells of filling;When higher than the first setting value, then it is judged as lymphocyte;
The image in NK cell edges is split again, the area and circularities of leucocyte iuuminting spot is calculated, works as height
When the second setting value, then it can determine that as NK cells.
2. NK cell detection methods according to claim 1, which is characterized in that obtain comprising the micrograph including NK cells
Picture:
Microscopic system obtains human body promoting blood circulation image;
Image is acquired by the video heads being fixed on microscopic system, after being converted into electric signal, is transmitted to analysis module.
3. NK cell detection methods according to claim 1, which is characterized in that thin containing light transmission spot and NK in leucocyte
The intramarginal image of born of the same parents is all to carry out image segmentation by the method for automatic correction threshold, improves the accuracy of edge detection.
4. a kind of NK cell detecting systems, which is characterized in that including:
Microscopic system, the microscopic system be used for obtains include NK cells including micro-image and obtain leucocyte again
Micro-image;
Image information collecting module, described image information acquisition module are connect with microscopic system, aobvious for receiving and handling
Micro- image;
Analysis module, the analysis module are connect with image information collecting module and control module, for analyzing the micro- of acquisition
Image;Edge detection is carried out, and fills edge internal voids, to form complete NK cell shapes;It is thin to calculate the front and back NK of filling
The difference in areas of born of the same parents;When higher than the first setting value, then it is judged as lymphocyte;The image in NK cell edges is divided again
It cuts, calculates the area and circularities of leucocyte iuuminting spot, when higher than the second setting value, then can determine that as NK cells;
Output module, the output module are connect with analysis module, the analysis result for exporting analysis module.
5. NK cell detecting systems according to claim 4, which is characterized in that the output module is printer.
6. NK cell detecting systems according to claim 4, which is characterized in that the microscopic system further includes can be real
Existing auto-translating and three axis of focusing move objective table automatically.
7. NK cell detecting systems according to claim 4, which is characterized in that the microscopic system include microscope and
Lens group is constituted, and the lens group is connect with microscope, and the video heads are fixed in lens group.
8. NK cell detecting systems according to claim 7, which is characterized in that the lens group it is glued by multiple lens and
At.
9. NK cell detecting systems according to claim 4, which is characterized in that further include control module, the control mould
On the one hand block is connect with analysis module, on the other hand connect with microscopic system.
10. NK cell detecting systems according to claim 4, which is characterized in that described image information acquisition module is number
Word video heads or analog video head.
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| CN201711416647.2A CN108693115A (en) | 2017-12-13 | 2017-12-13 | NK cell detection methods and system |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023042999A1 (en) * | 2021-09-17 | 2023-03-23 | 주식회사 메타이뮨텍 | Apparatus for analyzing natural killer cell activity using lens-free shadow imaging technique, and method therefor |
| CN117853850A (en) * | 2024-03-07 | 2024-04-09 | 威海紫光科技园有限公司 | Detection and evaluation system and method for NK cell cultivation process |
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| CN117853850A (en) * | 2024-03-07 | 2024-04-09 | 威海紫光科技园有限公司 | Detection and evaluation system and method for NK cell cultivation process |
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