CN101858908B - Method by utilizing magnetic fluorescent nanometer particle to mark housefly nerve cell - Google Patents
Method by utilizing magnetic fluorescent nanometer particle to mark housefly nerve cell Download PDFInfo
- Publication number
- CN101858908B CN101858908B CN 201010179854 CN201010179854A CN101858908B CN 101858908 B CN101858908 B CN 101858908B CN 201010179854 CN201010179854 CN 201010179854 CN 201010179854 A CN201010179854 A CN 201010179854A CN 101858908 B CN101858908 B CN 101858908B
- Authority
- CN
- China
- Prior art keywords
- housefly
- magnetic
- nanometer particle
- adds
- fluorescent
- 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.)
- Active
Links
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention belongs to the fluorescent marking cell technology, and relates to a method by utilizing magnetic fluorescent nanometer particle to mark a housefly nerve cell, which overcomes the weakness of the prior fluorescent marking technology that the fluorescein is easy to be dissociated from the protein to cause the difficult observation of the shape of the housefly under the fluorescent microscope. The method comprises the following steps: preparing magnetic fluorescent nanometer particle compound; preparing suspending solution of the housefly nerve cell: taking a live adult housefly to be soaked in 75 percent alcohol to be sterilized; cut a head of the housefly; the housefly with the head being cut off is arranged inside a pH neutral phosphorous acid buffering liquid to be made into the suspending liquid of housefly nerve cell, filtering the suspending liquid with a 80-mesh sieve to be used; adding double-antibody suspension; arranging the suspending liquid in a culture box to be hatched for 15 to 30 minutes; washing magnetic particle; and adding phosphorous acid buffering liquid to be shaken, film made and dried, and observing the shape of the cell under the fluorescent microscope. The method has the advantages that: the long-term stable luminescence of the fluorescein can be maintained; the separation efficiency of the received body nerve cell is high; and the shape observation under the fluorescent microscope is correct.
Description
Technical field
The invention belongs to the fluorescence labeled cell technology, specifically a kind of method of utilizing magnetic fluorescent nanometer particle to mark housefly nerve cell.
Background technology
Nanometer technology is progressively to set up beginning of the nineties late 1980s and the emerging technology that grows up, and has obtained developing rapidly in 21 century.Nanometer technology is counted as the three large important technologies that promote the 21 century development with biotechnology, infotech.The application prospect of nanometer technology has run through from the microscopic fields take atom, molecule as main body, microcosmic and the zone of transition of macroscopic view and the macrocosm of mankind's activity.Meanwhile, the multidisciplinary intersection on the nanoscale has represented the huge vitality of nanometer technology, and becoming rapidly one has extensively and the technical field of potential application foreground.1993, international nanotechnology steering committee was divided into 6 branches with nanometer technology: nanoelectronics, nanophysics, nanochemistry, nanoprocessing, Nanometrology and nanobiology.
Magnetic fluorescent nanometer particle not only has the characteristics such as the easy functionalization of good superparamagnetism and surface, and utilizes fluorescence directedly to identify and the target-marking material.The magnetic fluorescence particle surface introduce as-OH ,-COOH ,-CHO ,-the multiple reactive functional groups such as NH2, can also pass through the covalent bonds part, thereby make itself and biological acceptor specific binding under fluorescent microscope, observe its form.
At present, the fluorescein of fluorescence labeling utilization is fluorescein isothiocynate, phycoerythrin, texas Red, rhodamine etc.Prior art fluorescence labeling protein Marsshall method, thus utilizing the N=C=S bond of lysine on the protein and fluorescent dye to close to form compound sends fluorescence.The shortcoming of prior art is that the N=C=S bond of lysine and fluorescent dye is closed reaction and must be carried out under the alkali condition of pH8-9, in case environmental baseline and pH value change, fluorescein is easy to disintegrate down from protein, so that be difficult to accurately observe its form under fluorescent microscope.
Therefore invent a kind of magnetic fluorescent nanometer particle that utilizes and be combined with biological acceptor, under neutrallty condition, carry out fluorescence labeling; Keeping the luminous of fluorescein long-term stability, improve labeling effciency, is very important so that accurately observe the method for the utilizing magnetic fluorescent nanometer particle to mark housefly nerve cell of its form under fluorescent microscope.
Summary of the invention
The object of the present invention is to provide a kind of magnetic fluorescent nanometer particle that utilizes to be combined with biological acceptor, under neutrallty condition, carry out fluorescence labeling; Keep the luminous of fluorescein long-term stability, improve labeling effciency; So that under fluorescent microscope, accurately observe the method for the utilizing magnetic fluorescent nanometer particle to mark housefly nerve cell of its form.
The object of the present invention is achieved like this:
A kind of method of utilizing magnetic fluorescent nanometer particle to mark housefly nerve cell, step is as follows:
(1) preparation magnetic fluorescent nanometer particle compound:
A takes by weighing the surface and has modified amino magnetic fluorescent nanometer particle, adds the phosphate buffer of pH value 6.5-7.5, ultrasonic dispersion 5-15min;
B adds the N of 0.1-0.5mol/L, N-dicyclohexyl carbimide condensation agent in the steps A product; Hatch 10-25min for 37 ℃;
C is hatched 10-20min for 37 ℃ to the γ-aminobutyric acid solution of step B product adding 0.05-0.5mol/L;
D adds magnetic field with step C product, washs the magnetic grain 2 times with DMF and distilled water successively;
E adds penicillin and streptomysin mixed liquor with step D gained magnetic grain, and-20 ℃ save backup;
(2) preparation housefly neurocyte suspension:
A gets adult housefly alive, places 4 ℃ of refrigerators to place 5-8 minutes, immerses 75% alcohol disinfecting;
The housefly that B will sterilize lies on the surface plate, cuts head, the tiny cranium of collector;
C inserts the phosphate buffer of pH6.5-7.5 with the housefly head, makes behind the housefly neurocyte suspension by 80 mesh sieves, filtrate for later use;
(3) magnetic nano-particle mark housefly neurocyte:
A adds the two anti-suspensions of magnetic fluorescent nanometer particle compound of part to housefly neurocyte suspension; Place 37 ℃, saturated humidity, 5%CO
2Hatch 15-30min in the incubator;
B discards clear liquid under the magnetic field, uses the phosphate buffer of pH6.5-7.5 to wash the magnetic grain 2 times;
C to above-mentioned magnetic grain add pH6.5-7.5 phosphoric acid PBS damping fluid, shake up, film-making, dry up observation of cell form under the fluorescent microscope.
Main points of the present invention are preparation magnetic fluorescent nanometer particle compound, the part GABA receptor albumen by fluorescence labeling housefly neuronal cell surface under neutrallty condition and the neurocyte of mark housefly.
Magnetic fluorescent nanometer particle compound syntheti c route of the present invention is as shown below:
At first at N, amido modified magnetic fluorescent nanometer particle is combined with part γ-aminobutyric acid GABA and is formed compound under the N-dicyclohexyl carbimide DCC catalysis;
Then utilize the magnetic fluorescent nanometer particle compound mark and separate the neurocyte of housefly; Labeling process carries out in the phosphate buffer medium of pH=6.5-7.5.
The method of fluorescence labeled cell of the present invention is to realize fluorescence labeling to whole cell by the specific protein in fluorescence labeled cell film surface.
The concentration of the part γ-aminobutyric acid GABA that the present invention uses is 0.05-0.5mol/L; N, the concentration of N-dicyclohexyl carbimide condensation agent DCC is 0.1-0.5mol/L; The particle incubation time is 10-30min.
The present invention adds penicillin and streptomysin mixed liquor to magnetic grain nano particle, is to preserve for safety asepsis.
Characteristics of the present invention are:
(1) the magnetic fluorescent nanometer particle compound surface of the present invention preparation behind amino functional with γ-aminobutyric acid GABA ligand binding, according to the γ-aminobutyric acid GABA receptors bind of part and biological acceptor specific binding principle and housefly neuronal cell surface.
(2) this fluorescence labeling protein process is carried out under neutrallty condition.
(3) this magnetic fluorescent nanometer particle compound can be specific in conjunction with the neurocyte of release surface with the housefly of γ-aminobutyric acid GABA acceptor, and separation efficiency is high.
Advantage of the present invention is:
1, neutrallty condition fluorescence labeling can keep the fluorescein long-term stability luminous;
2, acceptor neurocyte, separation efficiency is high;
3, improve labeling effciency, morphologic observation is accurate under the fluorescent microscope.
Description of drawings
Fig. 1 is the housefly neurocyte figure of Mingguang City's state magnetic fluorescent nanometer particle to mark.
Fig. 2 is the housefly neurocyte figure of fluorescence excitation state magnetic fluorescent nanometer particle to mark.
Fig. 3 is the atomic force microscope figure of magnetic fluorescent nanometer particle compound.
Fig. 4 is the transmission electron microscope picture of magnetic fluorescent nanometer particle compound.
Fig. 5 is vibrating sample magnetometer (VSM) the test result figure of magnetic fluorescent nanometer particle compound.
Embodiment
The present invention will be further described below by embodiment, but the present invention is not subjected to the restriction of embodiment.
Embodiment 1:
Prepare amido modified magnetic fluorescent nanometer particle compound.
Take by weighing a certain amount of surface and modified amino magnetic fluorescent nanometer particle MFNP-NH
2The phosphoric acid PBS damping fluid that adds a small amount of pH=7.4, the N that adds 0.2mol/L behind the ultrasonic dispersion 5min, N-dicyclohexyl carbimide condensation agent DCC, 37 ℃ hatch 10min after, the γ-aminobutyric acid solution GABA solution that adds 0.2mol/L is hatched behind the 15min under the magnetic field successively with DMF and distilled water washing magnetic grain 2 times for 37 ℃; The magnetic grain adds penicillin and streptomysin mixed liquor, and-20 ℃ of preservations are stand-by.
Get adult housefly alive, place 4 ℃ of refrigerators to place to be immersed in after several minutes 75% alcohol to sterilize, carefully housefly is lain on the surface plate and decaptitate.The tiny cranium of collector toward the phosphoric acid PBS damping fluid that wherein adds pH=7.4, is made behind the suspension by 80 purpose sieves, and filtrate is stand-by.
Get the two anti-suspensions of magnetic fluorescent nanometer particle compound that housefly neurocyte suspension adds a small amount of part, place 37 ℃, saturated humidity, 5%CO
2Hatch 15min in the incubator, discard the stillness of night under the magnetic field, add a small amount of phosphoric acid PBS damping fluid with behind the phosphoric acid PBS damping fluid of the pH=7.4 washing magnetic grain 2 times, shake up, film-making dries up observation of cell form under the rear fluorescent microscope, sees Fig. 1.
The result shows: the surface has the neurocyte of the housefly of γ-aminobutyric acid solution GABA acceptor to be separated by magnetic fluorescent nanometer particle and is marked, and there are no unmarked neurocyte, sees Fig. 1 under Mingguang City; Show that the separation marking probability is 100%.The neurocyte of as seen being combined with MNFP under fluorescent microscope contains the red fluorescence element in because of particle and shows redness.See Fig. 2; Be neuronal cell on the cellular morphology that is labeled out, cell outline is clear, can see obvious neurocyte aixs cylinder structure.
Embodiment 2:
Take by weighing a certain amount of surface and modified amino magnetic fluorescent nanometer particle (MFNP-NH
2), the phosphoric acid PBS damping fluid that adds a small amount of pH=6.8, the N that adds 0.5mol/L behind the ultrasonic dispersion 15min, N-dicyclohexyl carbimide condensation agent DCC, hatch the γ-aminobutyric acid solution GABA solution that adds 0.5mol/L behind the 20min for 37 ℃, hatch behind the 20min under the magnetic field successively with DMF and distilled water washing magnetic grain for 37 ℃.The magnetic grain adds penicillin and streptomysin mixed liquor, and-20 ℃ of preservations are stand-by.
Get adult housefly alive, place 4 ℃ of refrigerators to place to be immersed in after several minutes 75% alcohol to sterilize, carefully housefly is lain on the surface plate and decaptitate.The tiny cranium of collector toward the phosphoric acid PBS damping fluid that wherein adds pH=7.0, is made behind the suspension by 80 purpose sieves, and filtrate is stand-by.
Get the two anti-suspensions of magnetic fluorescent nanometer particle compound that housefly neurocyte suspension adds a small amount of part, place 37 ℃, saturated humidity, 5%CO
2Hatch 20min in the incubator, discard the stillness of night under the magnetic field, behind the phosphoric acid PBS damping fluid of pH=7.0 washing magnetic grain 3 times, add a small amount of phosphoric acid PBS damping fluid, shake up, film-making dries up observation of cell form under the rear fluorescent microscope.Separation efficiency 80%.
Embodiment 3:
Take by weighing a certain amount of surface and modified amino magnetic fluorescent nanometer particle (MFNP-NH
2), the phosphoric acid PBS damping fluid that adds a small amount of pH=7.5, the N that adds 0.1mol/L behind the ultrasonic dispersion 15min, N-dicyclohexyl carbimide condensation agent DCC, hatch the γ-aminobutyric acid solution GABA solution that adds 0.5mol/L behind the 25min for 37 ℃, hatch behind the 10min under the magnetic field successively with DMF and distilled water washing magnetic grain for 37 ℃.The magnetic grain adds penicillin and streptomysin mixed liquor, and-20 ℃ of preservations are stand-by.
Get adult housefly alive, place 4 ℃ of refrigerators to place to be immersed in after several minutes 75% alcohol to sterilize, carefully housefly is lain on the surface plate and decaptitate.The tiny cranium of collector toward the phosphoric acid PBS damping fluid that wherein adds pH=7.5, is made behind the suspension by 80 purpose sieves, and filtrate is stand-by.
Get the two anti-suspensions of magnetic fluorescent nanometer particle compound that housefly neurocyte suspension adds a small amount of part, place 37 ℃, saturated humidity, 5%CO
2Hatch 30min in the incubator, discard the stillness of night under the magnetic field, add a small amount of phosphoric acid PBS damping fluid with behind the phosphoric acid PBS damping fluid of the pH=7.5 washing magnetic grain 3 times, shake up, film-making dries up observation of cell form under the rear fluorescent microscope.Separation efficiency 90%.
Embodiment 4:
Take by weighing a certain amount of surface and modified amino magnetic fluorescent nanometer particle (MFNP-NH
2), the phosphoric acid PBS damping fluid that adds a small amount of pH=6.5, the N that adds 0.3mol/L behind the ultrasonic dispersion 15min, N-dicyclohexyl carbimide condensation agent DCC, hatch the γ-aminobutyric acid solution GABA solution that adds 0.1mol/L behind the 20min for 37 ℃, hatch behind the 20min under the magnetic field successively with DMF and distilled water washing magnetic grain for 37 ℃.The magnetic grain adds penicillin and streptomysin mixed liquor, and-20 ℃ of preservations are stand-by.
Get adult housefly alive, place 4 ℃ of refrigerators to place to be immersed in after several minutes 75% alcohol to sterilize, carefully housefly is lain on the surface plate and decaptitate.The tiny cranium of collector toward the phosphoric acid PBS damping fluid that wherein adds pH=6.5, is made behind the suspension by 80 purpose sieves, and filtrate is stand-by.
Get the two anti-suspensions of magnetic fluorescent nanometer particle compound that housefly neurocyte suspension adds a small amount of part, place 37 ℃, saturated humidity, 5%CO
2Hatch 30min in the incubator, discard the stillness of night under the magnetic field, behind the phosphoric acid PBS damping fluid of pH=6.5 washing magnetic grain 2 times, add a small amount of phosphoric acid PBS damping fluid, shake up, film-making dries up observation of cell form under the rear fluorescent microscope.Separation efficiency 90%.
Embodiment 5:
Take by weighing a certain amount of surface and modified amino magnetic fluorescent nanometer particle (MFNP-NH
2), the phosphoric acid PBS damping fluid that adds a small amount of pH=7.2, the N that adds 0.2mol/L behind the ultrasonic dispersion 15min, N-dicyclohexyl carbimide condensation agent DCC, hatch the γ-aminobutyric acid solution GABA solution that adds 0.05mol/L behind the 15min for 37 ℃, hatch behind the 20min under the magnetic field successively with DMF and distilled water washing magnetic grain for 37 ℃.The magnetic grain adds penicillin and streptomysin mixed liquor, and-20 ℃ of preservations are stand-by.
Get adult housefly alive, place 4 ℃ of refrigerators to place to be immersed in after several minutes 75% alcohol to sterilize, carefully housefly is lain on the surface plate and decaptitate.The tiny cranium of collector toward the phosphoric acid PBS damping fluid that wherein adds pH=7.2, is made behind the suspension by 80 purpose sieves, and filtrate is stand-by.
Get the two anti-suspensions of magnetic fluorescent nanometer particle compound that housefly neurocyte suspension adds a small amount of part, place 37 ℃, saturated humidity, 5%CO
2Hatch 20min in the incubator, discard the stillness of night under the magnetic field, add a small amount of phosphoric acid PBS damping fluid with behind the phosphoric acid PBS damping fluid of the pH=7.2 washing magnetic grain 3 times, shake up, film-making dries up observation of cell form under the rear fluorescent microscope; Separation efficiency 88%.
Embodiment 6:
Implementation result:
The magnetic fluorescent nanometer particle compound of the part that several steps are processed among the process embodiment 1, through atomic force microscope and transmission electron microscope observing, the particle diameter of measuring magnetic fluorescent nanosphere is relatively even, and particle surface is bright and clean, can be used as carrier and probe and is used for research; See Fig. 3, Fig. 4.
Embodiment 7:
Implementation result:
The magnetic fluorescent nanometer particle compound of the part that several steps are processed among the process embodiment 3, through vibrating sample magnetometer (VSM) test, magnetic effect is good, can be used for magnetic and separates.See Fig. 5.
Embodiment 8:
Implementation result:
The magnetic fluorescent nanometer particle compound of the part that several steps are processed among the process embodiment 4, through vibrating sample magnetometer (VSM) test, magnetic effect is good, can be used for magnetic and separates.
The above is the preferred embodiments of the present invention only, is not limited to the present invention, and for a person skilled in the art, the present invention can have change and change.Within the spirit and principles in the present invention all, any modification of doing, improvement etc. all should be included within protection scope of the present invention.
Claims (1)
1. the method for a utilizing magnetic fluorescent nanometer particle to mark housefly nerve cell, step is as follows:
(1) preparation magnetic fluorescent nanometer particle compound:
A takes by weighing the surface and has modified amino magnetic fluorescent nanometer particle, adds the phosphate buffer of pH value 6.5-7.5, ultrasonic dispersion 5-15min;
Amino magnetic fluorescent nanometer particle preparation method has been modified on described surface: be that raw material prepares magnetic fluorescent nanometer particle with magnetic nano-particle, TEOS and RB200, process through N-(2-ethylamino-)-3-aminopropyl trimethyl silane again, obtain the surface and modified amino magnetic fluorescent nanometer particle;
B adds the N of 0.1-0.5mol/L, N-dicyclohexyl carbimide condensation agent in the steps A product; Hatch 10-25min for 37 ℃;
C is hatched 10-20min for 37 ℃ to the γ-aminobutyric acid solution of step B product adding 0.05-0.5mol/L;
D adds magnetic field with step C product, washs the magnetic grain 2 times with DMF and distilled water successively;
E adds penicillin and streptomysin mixed liquor with step D gained magnetic grain, and-20 ℃ save backup;
(2) preparation housefly neurocyte suspension:
A gets adult housefly alive, places 4 ℃ of refrigerators to place 5-8 minutes, immerses 75% alcohol disinfecting;
The housefly that B will sterilize lies on the surface plate, cuts head, the tiny cranium of collector;
C inserts the phosphate buffer of pH6.5-7.5 with the housefly head, makes behind the housefly neurocyte suspension by 80 mesh sieves, filtrate for later use;
(3) magnetic nano-particle mark housefly neurocyte:
A adds the two anti-suspension of the magnetic fluorescent nanometer particle compound that is connected with part to housefly neurocyte suspension; Place 37 ℃, saturated humidity, 5%CO
2Hatch 15-30min in the incubator;
B discards clear liquid under the magnetic field, uses the phosphate buffer of pH6.5-7.5 to wash the magnetic grain 2 times;
C to above-mentioned magnetic grain add pH6.5-7.5 phosphoric acid PBS damping fluid, shake up, film-making, dry up observation of cell form under the fluorescent microscope.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010179854 CN101858908B (en) | 2010-05-21 | 2010-05-21 | Method by utilizing magnetic fluorescent nanometer particle to mark housefly nerve cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010179854 CN101858908B (en) | 2010-05-21 | 2010-05-21 | Method by utilizing magnetic fluorescent nanometer particle to mark housefly nerve cell |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101858908A CN101858908A (en) | 2010-10-13 |
| CN101858908B true CN101858908B (en) | 2013-05-29 |
Family
ID=42944928
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201010179854 Active CN101858908B (en) | 2010-05-21 | 2010-05-21 | Method by utilizing magnetic fluorescent nanometer particle to mark housefly nerve cell |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101858908B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1888007A (en) * | 2006-07-21 | 2007-01-03 | 上海师范大学 | Nanometer magnetic fluorescent microsphere and its prepn and application |
| CN101235187A (en) * | 2008-03-05 | 2008-08-06 | 南京大学 | Magnetic fluorescence nano composite material, preparation method and application thereof |
| CN100505118C (en) * | 2007-03-09 | 2009-06-24 | 上海师范大学 | A making method for biologically compatible magnetic fluorescence nano particle |
-
2010
- 2010-05-21 CN CN 201010179854 patent/CN101858908B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1888007A (en) * | 2006-07-21 | 2007-01-03 | 上海师范大学 | Nanometer magnetic fluorescent microsphere and its prepn and application |
| CN100505118C (en) * | 2007-03-09 | 2009-06-24 | 上海师范大学 | A making method for biologically compatible magnetic fluorescence nano particle |
| CN101235187A (en) * | 2008-03-05 | 2008-08-06 | 南京大学 | Magnetic fluorescence nano composite material, preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101858908A (en) | 2010-10-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109507418B (en) | Magnetic nanoparticle with cell-like structure, immunomagnetic nanoparticle, and preparation method and application thereof | |
| Machałowski et al. | Spider Chitin. The biomimetic potential and applications of Caribena versicolor tubular chitin | |
| CN106198963A (en) | A kind of for immunomagnetic beads capturing leukocyte and preparation method thereof | |
| CN102732475A (en) | Microcarrier used for cell culture, its preparation method and its detection method | |
| CN106405075A (en) | Immunomagnetic bead and preparation method thereof | |
| CN104480101B (en) | Preparation of magnetic nano particle immobilized amino-acylase and products thereof and application | |
| CN107354134B (en) | Target cell capture substrate of nanorod array and preparation method and application thereof | |
| CN106399251A (en) | Antibody-coupled bionic immune magnetic sphere and preparation method thereof | |
| CN106567252B (en) | Fiber carrier and its preparation method and application | |
| CN106699904A (en) | Method for preparing hyperbranched cellulose nanocrystal flocculation material by mixed acid alternate hydrolysis | |
| CN105606684B (en) | A kind of graphene based on protein-single-walled carbon nanotube-nano-Au composite preparation method and applications | |
| CN109593523A (en) | A kind of preparation method and application of hypha,hyphae nitrogen sulphur auto-dope carbon dots | |
| CN109593719A (en) | A method of ferric oxide nano particles are adsorbed on cell membrane | |
| CN1920012A (en) | Reagent box for separating karyocyte in vitro and application method thereof | |
| CN104826126A (en) | Method for preparing high mechanical strength three-component fluorescence hydrogel | |
| Zhu et al. | Observing Thermobifida fusca cellulase binding to pretreated wood particles using time-lapse confocal laser scanning microscopy | |
| CN101858908B (en) | Method by utilizing magnetic fluorescent nanometer particle to mark housefly nerve cell | |
| KR101347205B1 (en) | Enzyme immobilized on Au-doped Magnetic silica nanoparticle, method for producing the same, and hydrolytic degradation method of biomass thereby | |
| WO2016113933A1 (en) | Cellulose nano-structure and method for producing same | |
| CN109880619A (en) | A method of Fe (III) ion is detected using the biomass-based carbon quantum dot of N doping | |
| CN111678900A (en) | Cell membrane and cell wall fluorescence labeling method based on tyrosinase catalysis and application | |
| CN103207202A (en) | Paramagnetic nano Fe-Ni alloy probe based quick detecting method for NMR (nuclear magnetic resonance) food-borne pathogenic bacteria | |
| CN108996530A (en) | A kind of preparation method for nanometer magnesium oxide and a kind of nano magnesia-nanofiber composite felt and its preparation method and application | |
| CN106179176B (en) | A kind of Mn3O4Nano-magnetic aeroge sorbing material and its preparation method and application | |
| CN109111499A (en) | A method of extracting ground bacillus Extracellular polymers |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |