CN108225875A - The preparation of nasal cavity washing liquid excretion body, identification method - Google Patents
The preparation of nasal cavity washing liquid excretion body, identification method Download PDFInfo
- Publication number
- CN108225875A CN108225875A CN201810033509.4A CN201810033509A CN108225875A CN 108225875 A CN108225875 A CN 108225875A CN 201810033509 A CN201810033509 A CN 201810033509A CN 108225875 A CN108225875 A CN 108225875A
- Authority
- CN
- China
- Prior art keywords
- excretion body
- liquid
- nasal cavity
- washing liquid
- cavity washing
- 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
Classifications
-
- 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
- G01N1/34—Purifying; Cleaning
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Pathology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- General Physics & Mathematics (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Urology & Nephrology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The nasal cavity washing preparation of liquid excretion body, identification method the invention discloses a kind of non-diagnostic purposes, the presence including the nasal cavity washing acquisition of liquid, the extraction and purification of excretion body, Electronic Speculum identification excretion body, immune-blotting method PROTEIN C D9, CD63.The method of the present invention simple possible, easily and effectively realizes the extraction of excretion body in nasal cavity washing liquid；Accurately and reliably, overcoming existing technology of preparing, there are background impurities interference, and the excretion body rate of recovery is low, identifies the deficiencies of albumen is few for the method for the present invention.Moreover, the method for the present invention can be directly used in the advantage of subsequent detection, there are the application values such as higher scientific research.
Description
Technical field
Preparation, identification method the present invention relates to a kind of nasal cavity washing liquid excretion body.
Background technology
Important tool of the excretion body as cell and intercellular communication, be invaginated by intracellular lysosome particle formed it is more
Foam, under specific stimulation, and cell membrane fusion, to the vesicles of a diameter of 50-100nm of exocytosis.Excretion
Body is widely present in tissue and body fluid, by transporting the bioactive substances such as protein, mRNA, miRNA that it is rich in, ginseng
Face cytoproximal signal path etc. with immunological regulation, the reconstruct of regulating cell epimatrix, activation, there is weight in scientific research etc.
The directive function wanted.
Nasal cavity washes one's hands and face liquid（nasal lavage fluid, NLF）A kind of body containing biological information as nasal cavity part
Liquid because its access approaches has the characteristics that non-intruding, safe and simple, has broad application prospects.
At present, there are extracting method is cumbersome, extraction efficiency is relatively low, background interference for the extracting method of nasal cavity washing liquid excretion body
The deficiencies of serious, and there is no unified standard step.
Invention content
The purpose of the present invention is to provide preparation, the identifications of nasal cavity washing liquid excretion body that a kind of method is easy, effect is good
Method.
The present invention technical solution be：
A kind of preparation of the nasal cavity washing liquid excretion body of non-diagnostic purposes, identification method, it is characterized in that：Include the following steps：
（One）Nasal cavity washes one's hands and face the acquisition of liquid
Healthy volunteer and patients with chronic sinusitis nasal cavity washing liquid are acquired respectively.Specific operation process is as follows：By 5 milliliters of physiology
Brine instils in the left nose hole of subject, and head is advised to be tilted backwards in 30 °, and soft palate is closed, and then head turns forward, and collects nose
Chamber washes one's hands and face liquid；Right nostril repeats the above steps；
（Two）The extraction and purification of excretion body
1. the nasal cavity washing liquid of collection is filtered by strainer, preliminary impurity screening and bacterium；
2. filtered nasal cavity washing liquid centrifuge 10 minutes under 4 DEG C, the centrifugal condition of 3000g, separation removal nasal cavity is washed one's hands and face
Cell and other impurities in liquid, then obtain supernatant liquid；
3. 2. supernatant liquid that step is obtained is moved into the second centrifuge tube, 30 points are centrifuged under 4 DEG C, the centrifugal condition of 6000g
Clock, further separating and removing impurities, obtains supernatant liquid；
4. 3. supernatant liquid that step is obtained is moved into third centrifuge tube, centrifugation 1 is small under 4 DEG C, the centrifugal condition of 10000g
When, obtain supernatant liquid；
5. surpassing the immigration of 4. supernatant liquid that step obtains from pipe, centrifuged 1 hour under 4 DEG C, the centrifugal condition of 10,0000g,
After removing supernatant liquid, precipitated；
6. after 5. precipitation that step obtains is resuspended washing with PBS, it is further purified by molecular exclusion chromatography；
7. surpassing the washing liquid immigration of step 6. treated nasal cavity from pipe, centrifuged again under 4 DEG C, the centrifugal condition of 10,0000g
1 hour, obtain being deposited in total excretion body of the nasal cavity washing liquid of the centrifugation bottom of the tube；
（Three）Electronic Speculum identifies excretion body
1. above-mentioned obtained excretion body is resuspended with PBS solution；
2. 20mL/L, pH6.8 Salkowski's solution is added dropwise, 1:1 mixing；
3. liquid takes above-mentioned mixing liquid 5ul drops on load sample copper mesh, at room temperature negative staining 10 minutes, dried under incandescent lamp；
4. transmission electron microscope observing is found：Products therefrom is the bilayer membrane structure foam of 50-100nm sizes；
（Four）The presence of immune-blotting method PROTEIN C D9, CD63
1. 1 × PBS for the excretion body 1ml that extraction purification is obtained is resuspended, excretion weight suspension is obtained, adds in 1ml lysates,
Extract total protein；
2. BCA methods measure excretion body protein content；
3. making 10% and 4% gradient glue, enough buffer solutions are added in, the excretion body total protein per hole loading 50ug；
4. installing electrode, connect with the mains, constant pressure 80V makes protein go on same starting line, and protein is run to separation gel
Shi Gaiwei 120V；
5. cutting glue, it is put into transferring film buffer solution；Pvdf membrane placement is polarized in methyl alcohol, is put into transferring film liquid in distilled water after aquation,
With 15 min are balanced together with cotton pad and filter paper in transferring film buffer solution；
6. pvdf membrane, glue, cotton pad and filter paper is fabricated to " sandwich " structure, first layer is cotton pad, and the second layer is filter paper, third
Layer is glue, and the 4th layer is pvdf membrane, and layer 5 is filter paper, and layer 6 is cotton pad, is clamped；It is put into transferring film slot, adds in transferring film liquid
And ice chest, transferring film slot are placed in the basin for filling ice water；Installation electrode connects power supply, and transferring film condition is set as 200 mA of constant current, when
Between 90 min；
7. after transferring film, pvdf membrane is taken out, is marked, is placed on shaking table, TBST washes 5 min × 3 time, 5 % degreasing oxen
Milk closes 1-2h；TBST washes 5 min × 3 time of film；Primary antibody CD9, CD63 is added to be incubated, is stayed overnight for 4 DEG C after 1 h of room temperature；
8. TBST washes 10 min × 3 time of film；Secondary antibody is incubated, 2 h of room temperature；TBST washes 20 min × 3 time；
9. developing in darkroom, X photographic films are rinsed；
Through the expression that distinctive PROTEIN C D9, CD63 in excretion body surface face is detected in above method products therefrom.
Excretion body is further identified using nano particle tracer technique, analysis has detected the excretion body that extraction purification obtains
Concentration, size are consistent with the characterization of excretion body.
The method of the present invention simple possible, easily and effectively realizes the extraction of excretion body in nasal cavity washing liquid；The method of the present invention
Accurately and reliably, overcoming existing technology of preparing, there are background impurities interference, and the excretion body rate of recovery is low, identifies the deficiencies of albumen is few.
Moreover, the method for the present invention can be directly used in the advantage of subsequent detection, there are the application values such as higher scientific research.
Description of the drawings
The invention will be further described with reference to the accompanying drawings and examples.
Fig. 1 is the electron micrograph that the present invention implements products therefrom.
Fig. 2 is to implement the immune-blotting method result that PROTEIN C D9, CD63 in products therefrom is expressed to the present invention.
Fig. 3 is that nano particle tracer technique analysis testing result is carried out to products therefrom of the present invention.
Specific embodiment
Not specified various instruments and reagent are commercial product well known in the art in the following example, can be passed through
Commercial sources obtain.
A kind of preparation of the nasal cavity washing liquid excretion body of non-diagnostic purposes, identification method, include the following steps：
First, the acquisition of nasal cavity washing liquid
Obtain participant know and the agreement of ethics under, respectively acquire healthy volunteer and patients with chronic sinusitis nasal cavity washing
Liquid.Specific operation process is as follows：5 milliliters of physiological saline are instiled in the left nose hole of subject, head is advised to be tilted backwards in 30 °,
Soft palate is closed, and then head turns forward, and collects nasal cavity washing liquid.Right nostril repeats the above steps.
2nd, the extraction and purification of excretion body
1. the nasal cavity washing liquid of collection is filtered by strainer, preliminary impurity screening and bacterium.
2. filtered nasal cavity washing liquid centrifuge 10 minutes under 4 DEG C, the centrifugal condition of 3000g, separation removes nasal cavity
The cell and other impurities in liquid are washed one's hands and face, then obtains supernatant liquid.
3. the supernatant liquid 2. obtained is moved into the second centrifuge tube, 30 points are centrifuged under 4 DEG C, the centrifugal condition of 6000g
Clock, further separating and removing impurities, obtains supernatant liquid.
4. the supernatant liquid 3. obtained is moved into third centrifuge tube, centrifugation 1 is small under 4 DEG C, the centrifugal condition of 10000g
When, obtain supernatant liquid.
5. surpassing the supernatant liquid 4. obtained immigration from pipe, centrifuged 1 hour under 4 DEG C, the centrifugal condition of 10,0000g,
After removing supernatant liquid, precipitated.
6. after the precipitation 5. obtained is resuspended washing with PBS, it is further purified by molecular exclusion chromatography.
7. surpassing the washing liquid immigration of 6. treated nasal cavity from pipe, centrifuged again under 4 DEG C, the centrifugal condition of 10,0000g
1 hour, obtain being deposited in total excretion body of the nasal cavity washing liquid of the centrifugation bottom of the tube.
3rd, Electronic Speculum identification excretion body
1. above-mentioned obtained excretion body is resuspended with PBS solution.
2. 20mL/L Salkowski's solutions (pH6.8) 1 are added dropwise:1 mixing.
3. liquid takes above-mentioned mixing liquid 5ul drops on load sample copper mesh, at room temperature negative staining 10 minutes, dried under incandescent lamp.
4. the photo that transmission electron microscope observing arrives as shown in Figure 1, it can be found that：Products therefrom divides for the double of 50-100nm sizes
The excretion body of sublayer membrane structure foam, morphology and other document reports is consistent.
4th, the presence of immune-blotting method PROTEIN C D9, CD63
1. the product (the excretion body extracted in nasal cavity washing liquid) of gained is resuspended with 1 × PBS of 1ml, excretion weight is obtained
Suspension adds in 1ml lysates, extracts total protein.
2. BCA methods measure excretion body protein content.
3. making 10% and 4% gradient glue, enough buffer solutions are added in, the excretion body total protein per hole loading 50ug.
4. installing electrode, connect with the mains, 80 V of constant pressure makes protein go on same starting line, protein run to point
120 V are changed to during from glue.
5. cutting glue, it is put into transferring film buffer solution.Pvdf membrane placement is polarized in methyl alcohol, is put into transferring film in distilled water after aquation
Liquid balances 15 min together with cotton pad and filter paper in transferring film buffer solution.
6. pvdf membrane, glue, cotton pad and filter paper are fabricated to " sandwich " structure, first layer is cotton pad, and the second layer is filter paper,
Third layer is glue, and the 4th layer is pvdf membrane, and layer 5 is filter paper, and layer 6 is cotton pad, is clamped.It is put into transferring film slot, adds in and turn
Film liquid and ice chest, transferring film slot are placed in the basin for filling ice water.Installation electrode connects power supply, and transferring film condition is set as constant current 200
MA, 90 min of time.
7. after transferring film, pvdf membrane is taken out, is marked, is placed on shaking table, TBST washes 5 min × 3 time, and 5 % take off
Fat milk closing 1-2 h.TBST washes 5 min × 3 time of film.Add primary antibody（CD9、CD63）It is incubated, is stayed overnight for 4 DEG C after 1 h of room temperature.
8. TBST washes 10 min × 3 time of film.Secondary antibody is incubated, 2 h of room temperature.TBST washes 20 min × 3 time.
9. developing in darkroom, X photographic films are rinsed.
Testing result is as shown in Fig. 2, through detecting the distinctive albumen in excretion body surface face in above method products therefrom
The expression of CD9, CD63, therefore, it is excretion body that products therefrom is illustrated from molecular characterization.
Nano particle tracer technique（NTA）Further identify excretion body
NTA is a kind of method of relatively new characterization nano particle, it can directly and in real time observe nano particle.NTA passes through
Light microscope collects the scattered light signal of nano particle, and one section of nano particle of shooting makees the image of Brownian movement in the solution,
The Brownian movement of each particle is tracked and analyzed, so as to calculate the hydrodynamic radius of nano particle and concentration.
In clinical diagnosis, quickly and easily under complicated biological context（Such as nasal cavity washing liquid, blood plasma, urine）It measures
Concentration, size and the characterize data of excretion body are indispensable requirements, presently, there are conventional method all can not perfectly solve this
One problem.NTA as a kind of completely new measuring technique, have real-time observation, higher resolution ratio, accurate measurement of concetration and
Fluorescence measurement function provides the new approaches to excretion body size and concentration studies.
Testing result is as shown in figure 3, NTA analyses have detected the concentration through above method products therefrom, size etc., with excretion
The characterization of body is consistent.
Claims (2)
1. a kind of preparation of the nasal cavity washing liquid excretion body of non-diagnostic purposes, identification method, it is characterized in that：Include the following steps：
（One）Nasal cavity washes one's hands and face the acquisition of liquid
Healthy volunteer and patients with chronic sinusitis nasal cavity washing liquid are acquired respectively；Specific operation process is as follows：By 5 milliliters of physiology
Brine instils in the left nose hole of subject, and head is advised to be tilted backwards in 30 °, and soft palate is closed, and then head turns forward, and collects nose
Chamber washes one's hands and face liquid；Right nostril repeats the above steps；
（Two）The extraction and purification of excretion body
1. the nasal cavity washing liquid of collection is filtered by strainer, preliminary impurity screening and bacterium；
2. filtered nasal cavity washing liquid centrifuge 10 minutes under 4 DEG C, the centrifugal condition of 3000g, separation removal nasal cavity is washed one's hands and face
Cell and other impurities in liquid, then obtain supernatant liquid；
3. 2. supernatant liquid that step is obtained is moved into the second centrifuge tube, 30 points are centrifuged under 4 DEG C, the centrifugal condition of 6000g
Clock, further separating and removing impurities, obtains supernatant liquid；
4. 3. supernatant liquid that step is obtained is moved into third centrifuge tube, centrifugation 1 is small under 4 DEG C, the centrifugal condition of 10000g
When, obtain supernatant liquid；
5. surpassing the immigration of 4. supernatant liquid that step obtains from pipe, centrifuged 1 hour under 4 DEG C, the centrifugal condition of 10,0000g,
After removing supernatant liquid, precipitated；
6. after 5. precipitation that step obtains is resuspended washing with PBS, it is further purified by molecular exclusion chromatography；
7. surpassing the washing liquid immigration of step 6. treated nasal cavity from pipe, centrifuged again under 4 DEG C, the centrifugal condition of 10,0000g
1 hour, obtain being deposited in total excretion body of the nasal cavity washing liquid of the centrifugation bottom of the tube；
（Three）Electronic Speculum identifies excretion body
1. above-mentioned obtained excretion body is resuspended with PBS solution；
2. 20mL/L, pH6.8 Salkowski's solution is added dropwise, 1:1 mixing；
3. liquid takes above-mentioned mixing liquid 5ul drops on load sample copper mesh, at room temperature negative staining 10 minutes, dried under incandescent lamp；
4. transmission electron microscope observing is found：Products therefrom is the bilayer membrane structure foam of 50-100nm sizes；
（Four）The presence of immune-blotting method PROTEIN C D9, CD63
1. 1 × PBS for the excretion body 1ml that extraction purification is obtained is resuspended, excretion weight suspension is obtained, adds in 1ml lysates,
Extract total protein；
2. BCA methods measure excretion body protein content；
3. making 10% and 4% gradient glue, enough buffer solutions are added in, the excretion body total protein per hole loading 50ug；
4. installing electrode, connect with the mains, constant pressure 80V makes protein go on same starting line, and protein is run to separation gel
Shi Gaiwei 120V；
5. cutting glue, it is put into transferring film buffer solution；Pvdf membrane placement is polarized in methyl alcohol, is put into transferring film liquid in distilled water after aquation,
With 15 min are balanced together with cotton pad and filter paper in transferring film buffer solution；
6. pvdf membrane, glue, cotton pad and filter paper is fabricated to " sandwich " structure, first layer is cotton pad, and the second layer is filter paper, third
Layer is glue, and the 4th layer is pvdf membrane, and layer 5 is filter paper, and layer 6 is cotton pad, is clamped；It is put into transferring film slot, adds in transferring film liquid
And ice chest, transferring film slot are placed in the basin for filling ice water；Installation electrode connects power supply, and transferring film condition is set as 200 mA of constant current, when
Between 90 min；
7. after transferring film, pvdf membrane is taken out, is marked, is placed on shaking table, TBST washes 5 min × 3 time, 5 % degreasing oxen
Milk closes 1-2h；TBST washes 5 min × 3 time of film；Primary antibody CD9, CD63 is added to be incubated, is stayed overnight for 4 DEG C after 1 h of room temperature；
8. TBST washes 10 min × 3 time of film；Secondary antibody is incubated, 2 h of room temperature；TBST washes 20 min × 3 time；
9. developing in darkroom, X photographic films are rinsed；
Through the expression that distinctive PROTEIN C D9, CD63 in excretion body surface face is detected in above method products therefrom.
2. the preparation of the nasal cavity washing liquid excretion body of non-diagnostic purposes according to claim 1, identification method, feature
It is：Excretion body is further identified using nano particle tracer technique, analysis have detected the concentration of excretion body that extraction purification obtains,
Size is consistent with the characterization of excretion body.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810033509.4A CN108225875A (en) | 2018-01-15 | 2018-01-15 | The preparation of nasal cavity washing liquid excretion body, identification method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810033509.4A CN108225875A (en) | 2018-01-15 | 2018-01-15 | The preparation of nasal cavity washing liquid excretion body, identification method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108225875A true CN108225875A (en) | 2018-06-29 |
Family
ID=62641797
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810033509.4A Pending CN108225875A (en) | 2018-01-15 | 2018-01-15 | The preparation of nasal cavity washing liquid excretion body, identification method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108225875A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101498734A (en) * | 2009-03-09 | 2009-08-05 | 昆明理工大学 | Method for simultaneously turning two films and marking protein TRX and procaspase12 |
| CN101782585A (en) * | 2010-02-10 | 2010-07-21 | 广州医学院 | Lung cancer tissue protein blotting membrane and preparation method thereof |
| CN102858997A (en) * | 2009-11-02 | 2013-01-02 | 新加坡科技研究局 | Methods for monitoring cellular states and for immortalizing mesenchymal stem cells |
| CN105779586A (en) * | 2015-12-28 | 2016-07-20 | 四川农业大学 | Method for separating exosomes from animal plasma and for detecting purity |
| CN108918228A (en) * | 2018-06-04 | 2018-11-30 | 北京全式金生物技术有限公司 | Excretion body in serum or blood plasma prepares kit and excretion preparation |
-
2018
- 2018-01-15 CN CN201810033509.4A patent/CN108225875A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101498734A (en) * | 2009-03-09 | 2009-08-05 | 昆明理工大学 | Method for simultaneously turning two films and marking protein TRX and procaspase12 |
| CN102858997A (en) * | 2009-11-02 | 2013-01-02 | 新加坡科技研究局 | Methods for monitoring cellular states and for immortalizing mesenchymal stem cells |
| CN101782585A (en) * | 2010-02-10 | 2010-07-21 | 广州医学院 | Lung cancer tissue protein blotting membrane and preparation method thereof |
| CN105779586A (en) * | 2015-12-28 | 2016-07-20 | 四川农业大学 | Method for separating exosomes from animal plasma and for detecting purity |
| CN108918228A (en) * | 2018-06-04 | 2018-11-30 | 北京全式金生物技术有限公司 | Excretion body in serum or blood plasma prepares kit and excretion preparation |
Non-Patent Citations (9)
| Title |
|---|
| CECILIA LASSER ET AL.: "RNA-containg exosomes in human nasal sections", 《AMERICAN JOURNAL OF RHINOLOGY & ALLERGY》 * |
| YIWEN YOU ET AL.: "Matrix metalloproteinase 13-containing exosomes promote nasopharyngeal carcinoma metastasis", 《CANCER SCI》 * |
| 张桂龙 等: "人源神经干细胞来源外泌体的提取鉴定及内吞作用", 《东南大学学报(医学报)》 * |
| 李卫芳 等: "《生物化学与分子生物学》", 31 January 2012 * |
| 李超 等: "胶质瘤源性exsome的提取及形态学观察", 《中国实验诊断学》 * |
| 渠香云 等: "一种新型血清外泌体分离方法的构建及其在乳腺癌微RNA检测中的应用", 《肿瘤》 * |
| 翁叶靖 等: "外泌体分离及其蛋白组学分析的研究进展", 《色谱》 * |
| 高佳佳 等: "血清饥饿对于结肠癌HCT116细胞外泌体分泌的影响", 《世界华人消化杂志》 * |
| 黄波 等: "K562细胞外泌体选择性携带MMP9基因", 《江西医药》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106093392B (en) | The integrated testing method and detection chip of a kind of urine excretion body separation, enrichment and detection | |
| CN108795869A (en) | A kind of circulating tumor cell positive enrichment method | |
| CN103424555B (en) | Reagent group for matching test before blood transfusion and test method thereof | |
| CN109628277A (en) | The separation of excretion in-vivo tumour mark miRNA a kind of and detection system and method | |
| CN105087493B (en) | Three kinds of monoclonal antibody coupling immunomagnetic beads of combination are applied in tumour cell sorting | |
| CN107449713A (en) | Method dependent on the circulating tumor cell sorting and enrichment of mixed antibody | |
| CN109486653A (en) | Trace cell capture system based on micro-fluidic and immune Magneto separate dual strategy | |
| CN109187978A (en) | It is a kind of detect circulating tumor cell HER2, ER, PR immunofluorescent reagent box and its application | |
| US20210170409A1 (en) | Microfluidic chip for circulating tumor cell separation, circulating tumor cell separation method and counting method | |
| Chang et al. | Biomimetic immunomagnetic gold hybrid nanoparticles coupled with inductively coupled plasma mass spectrometry for the detection of circulating tumor cells | |
| CN108795693A (en) | A kind of micro-fluidic chip of capture blood rare cell | |
| CN209485831U (en) | The extracellular vesica enriching apparatus of the urine of hospital | |
| JP7376021B2 (en) | Method for separating and analyzing microvesicles from human blood | |
| JP7444386B2 (en) | Method for isolating and analyzing microvesicles from human urine | |
| CN105910993A (en) | Household pocket-sized blood multi-parameter tester | |
| CN104359746B (en) | Cell mass collector and the method for collecting cell in humoral specimen | |
| JPH05501965A (en) | Method and apparatus for collecting amniotic fluid samples | |
| CN108225875A (en) | The preparation of nasal cavity washing liquid excretion body, identification method | |
| CN111351937A (en) | MMR protein expression deletion detection kit and detection method thereof | |
| CN109580951A (en) | The kit and its application method of multispecific antibody joint-detection early liver cancer marker | |
| CN209485830U (en) | The device of hospital and the home-use extracellular vesica of enrichment urine that can make | |
| CN218710571U (en) | Osteoporosis auxiliary diagnosis device based on exosome | |
| CN108795692A (en) | Rare cell capture systems and its application | |
| US20140309553A1 (en) | Kits and methods for separating a target analyte from a suspension | |
| JP2004279113A (en) | Method of manufacturing extract of antigen or antibody |
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 |
Application publication date: 20180629 |
|
| RJ01 | Rejection of invention patent application after publication |