CN108535222A - A kind of method of micro, quick measurement algae oil content - Google Patents
A kind of method of micro, quick measurement algae oil content Download PDFInfo
- Publication number
- CN108535222A CN108535222A CN201810165144.0A CN201810165144A CN108535222A CN 108535222 A CN108535222 A CN 108535222A CN 201810165144 A CN201810165144 A CN 201810165144A CN 108535222 A CN108535222 A CN 108535222A
- Authority
- CN
- China
- Prior art keywords
- algae
- concentration
- micro
- added
- content
- 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
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N15/1434—Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
-
- 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
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
Abstract
The invention discloses a kind of method of micro, quick measurement algae oil content, the method includes：(1) algae solution is taken, the number concentration of microalgae cell in algae solution is calculated；(2) dilute different concentration of algae algae solution；(3) mother liquor is prepared；(4) above-mentioned each concentration of algae takes 2 test tubes respectively, each HCl that 5 10mL algae solutions and 5 10ml1mol/L are added, after being sufficiently mixed, 0.1ul mother liquors are added in one branch pipe of each concentration of algae, the organic solvent in step (3) is added in another branch pipe, manages, mixes well as a contrast；(5) 400 600nm exciting lights of excitation wavelength are used, in fluorescence microscopy microscopic observation and photograph to record Unicell Oils and Fats cumulative change situation；(6) grease volume is calculated；(7) unit of account volume algae solution fat content.It is quick and easy using the method for the present invention detection.The present invention combines microdisk electrode and detection method together, and process is simple and convenient, i.e., measures microalgae grease content by In vivo detection, time more conventional method is needed to significantly reduce.
Description
Technical field
The present invention relates to microalgae field of biological energy source more particularly to a kind of methods of micro, quick measurement algae oil content.
Background technology
The progress of human civilization is largely dependent upon the development of the energy, at present to change in the energy used in the mankind
Based on stone fuel, and economic the increasing rapidly of civilization causes the mankind to rise the demand of the energy rapidly.World today's energy
Crisis is deepened, and the shortage of fossil fuel and its environmental problem brought are also increasingly prominent, therefore finds environmental-friendly renewable
The energy has become countries in the world problem of interest.Biodiesel as a kind of renewable, environmentally friendly novel energy
It is concerned, source is of greatest concern with microalgae, and reason is that microalgae has oil production height, the speed of growth soon and is easy to train
The advantages that supporting, therefore raw materials for production of the optimum as biodiesel.Based on this, microalgae biodiesel has become new energy neck
Research hotspot in domain develops for reducing using and ensureing that national energy security is of great significance for fossil fuel.
The measurement of the screening of oil-rich microalgae, culture and its fat content is to obtain the only way which must be passed of high quality algae, herein
In the process, it generally requires to carry out frequent grease quantitative for more microalgae sample, and for different cultivation period microalgaes oil
The tracking and measuring of fat is also more important.Traditional microalgae grease content assaying method is organic solvent extraction, that is, is weighed certain
Dry algae powder sample is measured, refluxing extraction is carried out using organic solvents such as ethers, n-hexane or chloroforms, extracting solution is collected and waits for that solvent is waved
It dries to constant weight after dry, according to the grease dry weight and algae powder Mass Calculation fat content finally obtained.Sample needed for this method
More (generally 2~5g of dry algae powder), organic solvent consumption is big and complex for operation step, required time is tediously long, easily because of the external world
Factor is interfered and generates error, therefore is unfavorable for system operatio.Emerging fluorescence spectrophotometry is to use fluorescent dye Buddhist nun sieve
It is red to be configured to certain density dye liquor, and the dye liquor is directly added into the microalgae algae solution of known concentration, it is incubated under dark condition
Its fluorescence intensity (fluorescence condition is measured after 5~10min：Excitation wavelength 480nm, 570~590nm of wavelength of transmitted light), then according to
The fluorescence intensity dyed according to calibration curve formula and sample algae solution calculates fat content.The method significantly reduce sample consumptions
Amount and minute, simple and easy to do but expensive fluorescent dye and detection device limit the application of this method in production,
In addition it needs the stronger dimethyl sulfoxide (DMSO) of toxicity (DMSO) or acetone as dyeing liquor solvent, is unfavorable for practical operation.Therefore,
Searching sample consumption is low, safe, low-cost fat content assay method swift to operate is of great significance.
Invention content
The main purpose of the present invention is to provide a kind of methods of micro, quick measurement algae oil content.
To achieve the above object, the method for a kind of micro, quick measurement algae oil content provided by the invention, the method
Include the following steps：
(1) algae solution for carrying out microdisk electrode is taken, the unit for calculating the number concentration C, C of microalgae cell in algae solution is cell/
ml；
(2) wet algae samples of the 0.5-1.0g through desalination point is weighed, 50-100ml distilled water is added, is sufficiently mixed uniformly, then
It is diluted to the algae solution of 1/2,1/3,1/4,1/5,1,/10 5 different concentration of algae；
(3) 0.4-0.8g fluorescent staining lipotropism fluorescent dyes powder is weighed, is dissolved in middle organic solvent, room temperature decentralization
It sets 4-6 days, during which often shakes, filtered after allowing it fully to dissolve, be configured to mother liquor；
(4) above-mentioned each concentration of algae takes 2 test tubes respectively, each HCl that 5-10mL algae solutions and 5-10ml1mol/L is added,
After being sufficiently mixed, 0.1ul mother liquors are added in one branch pipe of each concentration of algae, and the organic solvent in step (3) is added in another branch pipe, makees
For control tube, mix well；
(5) excitation wavelength 400-600nm exciting lights are used, in fluorescence microscopy microscopic observation and photograph to record single cell oil
Fat cumulative change situation；
(6) unicellular olesome number is recorded according to photo, and measures droplet diameter, calculated according to sphere volume formula
Each blob volume, each cell grease content are the adduction of all blob volumes；
(7) contained according to the number concentration C of microalgae cell and individual cells fat content unit of account volume algae solution grease
Amount：Vt=CV, C are the number concentration of microalgae cell, and V is Unicell Oils and Fats volume, and Vt is unit volume fat content.
Preferably, further include before the step (2)：：
(11) after repeating retarder thinner washing, the salinity of removal microalgae cell, centrifugation for several times, algae solution centrifugal concentrating.
Preferably, the retarder thinner in step (11) is distilled water, tap water, deionized water, ultra-clean water or mineral water.
Preferably, the fluorescent staining lipotropism fluorescent dyes be the Sudan three, Sudan IV or Sudan black B, it is described organic
Solvent is 100-200ml70% ethyl alcohol.
Preferably, the step (5) is specially：After microalgae cell dyeing under 507nm excitation wavelengths, swept using laser
It retouches Laser Scanning Confocal Microscope to microalgae cell tomoscan and to photograph to record every 0.01-0.5 μm, by 3D rendering composite software
Oil droplet 3D pictures are rebuild, olesome diameter is measured using microphotograph software.
The method of micro, the quick measurement algae oil content of the present invention includes that (1) takes the algae solution for carrying out microdisk electrode, calculates algae
The unit of the number concentration C, C of microalgae cell are cell/ml in liquid；(2) wet algae samples of the 0.5-1.0g through desalination point is weighed, is added
Enter 50-100ml distilled water, be sufficiently mixed uniformly, is then diluted to 1/2,1/3,1/4,1/5,1,/10 5 different concentration of algae
Algae solution；(3) 0.4-0.8g grams of fluorescent staining lipotropism fluorescent dyes powder is weighed, is dissolved in middle organic solvent, at room temperature
It places 4-6 days, during which often shakes, filtered after allowing it fully to dissolve, be configured to mother liquor；(4) above-mentioned each concentration of algae takes respectively
2 test tubes, each HCl that 5-10mL algae solutions and 5-10ml1mol/L is added, after being sufficiently mixed, one branch pipe of each concentration of algae is added
0.1ul mother liquors, another branch pipe are added the organic solvent in step (3), manage, mix well as a contrast；(5) excitation wave is used
Long 400-600nm exciting lights in fluorescence microscopy microscopic observation and photograph to record Unicell Oils and Fats cumulative change situation；(6) basis
Photo records unicellular olesome number, and measures droplet diameter, calculates each blob volume according to sphere volume formula, often
A cell grease content is the adduction of all blob volumes；(7) according to the number concentration C of microalgae cell and individual cells grease
Content calculation unit volume algae solution fat content：Vt=CV, C are the number concentration of microalgae cell, and V is Unicell Oils and Fats volume,
Vt is unit volume fat content.
1. detection is quick and easy.The present invention combines microdisk electrode and detection method together, and process is simple and convenient, i.e., logical
It crosses In vivo detection and measures microalgae grease content.Time more conventional method is needed to significantly reduce, single sample detection only needs 2 points
Clock.
V=π d1 3/6+πd2 3/6+…πdn 3/6
2. can the almost quantitative content for detecting unit volume grease.Fluorescence microscope simultaneously measures oil droplet number (n)
After droplet diameter (d), using the blob volume for calculating single microalgae, to by growth curve unit of account culture solution
The total amount of grease.
3. at low cost.Detection process does not need high-end instrument and equipment and consumptive material, therefore more conventional method is of low cost.
Specific implementation mode
It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to limit this hair
It is bright.
Embodiment：
(1) algae solution for carrying out microdisk electrode is taken, the unit for calculating the number concentration C, C of microalgae cell in algae solution is cell/
ml；
(11) after repeating retarder thinner washing, the salinity of removal microalgae cell, centrifugation for several times, algae solution centrifugal concentrating；Its
In, the retarder thinner is distilled water, tap water, deionized water, ultra-clean water or mineral water
(2) wet algae samples of the 0.5-1.0g through desalination point is weighed, 50-100ml distilled water is added, is sufficiently mixed uniformly, then
It is diluted to the algae solution of 1/2,1/3,1/4,1/5,1,/10 5 different concentration of algae；
(3) 0.4-0.8g grams of fluorescent staining lipotropism fluorescent dyes powder the Sudan three is weighed, 100-200ml70% is dissolved in
In ethyl alcohol, places 4-6 days at room temperature, during which often shake, filtered after allowing it fully to dissolve, be configured to mother liquor；
(4) above-mentioned each concentration of algae takes 2 test tubes respectively, each HCl that 5-10mL algae solutions and 5-10ml1mol/L is added,
After being sufficiently mixed, 0.1ul mother liquors are added in one branch pipe of each concentration of algae, and the organic solvent in step (3) is added in another branch pipe, makees
For control tube, mix well；
(5) excitation wavelength 400-600nm exciting lights are used, in fluorescence microscopy microscopic observation and photograph to record single cell oil
Fat cumulative change situation；
(6) unicellular olesome number is recorded according to photo, and measures droplet diameter, calculated according to sphere volume formula
Each blob volume, each cell grease content are the adduction of all blob volumes；
(7) contained according to the number concentration C of microalgae cell and individual cells fat content unit of account volume algae solution grease
Amount：Vt=CV, C are the number concentration of microalgae cell, and V is Unicell Oils and Fats volume, and Vt is unit volume fat content.
Specific experiment method：
1, under above-mentioned condition of culture, logarithmic growth phase algae is seeded to above-mentioned training as experiment material for experimental design
In nutrient solution, Initial seeding density is 2.375 ± 0.916 (× 104cell/ml).3 Duplicate Samples are set, it is bent to measure Growth of Platymonas Spp
Line and oil and fat accumulation situation.
2, growth measurement takes 1mL algae solutions every 2d from each experimental group, and 1~2 drop Lugol's is added to fix frustule, with
Blood counting chamber counts frustule under the microscope.Each sample count 3 times, 3 count results compare standard deviation not
More than 20%, the 4th counting is otherwise carried out.
3, light dyeing lipotropism fluorescent dyes the Sudan three is dissolved in organic solvent 100-200ml70% ethyl alcohol, prepares
The mother liquor of 10mM/L.In use, 0.1 μ L mother liquors is taken to inject dyeing 1min or so in 1mL algae solutions.Use fluorescence microscope
(NikonEclipse80i) 6 visuals field are randomly selected at 507nm to observe and photograph to record intracellular fat particles distribution feelings
Condition.
V=π d1 3/6+πd2 3/6+…πdn 3/6
4, after the dyeing of fat content measurement frustule laser scanning co-focusing microscope is used under 507nm excitation wavelengths
(NikonECLIPSETE2000-U) it to frustule tomoscan and is photographed to record every 0.5 μm, by Nikon EZ-C1 softwares
Processing system rebuilds oil droplet 3D pictures and shows the approximate positive sphere of oil droplet.Oil droplet is measured using Nikon NIS-elementsBR softwares
Particle diameter calculates each blob volume according to sphere volume formula, and each cell grease content is adding for all blob volumes
With：(wherein d is droplet diameter, and n is unicellular oil droplet number).It is calculated according to Growth of Platymonas Spp curve and individual cells fat content
Unit volume algae solution fat content：(C is flat algae cell concentration to Vt=CV, and V is Unicell Oils and Fats volume, Vt unit bodies long-pending oils
Fat content).
4) data method tests all data and is represented as mean+SD (mean ± SD), using SPSS 17.0
Statistical software carries out the one-factor analysis of variance (One-WayANOVA) Multiple range test analysis (LSDANOVA), P ＜ 0.05 between group
For the significance level of difference.
Wherein, the microalgae in the present embodiment includes schizochytrium, Isochrysis galbana, the hidden dinoflagellate of bandit's formula, micro- green ball
Algae, pavlova viridis, Botryococcus braunii, Phaeodactylum tricornutum, diamond shape algae, chlorella, scenedesmus or Du Shi algaes.
It these are only the preferred embodiment of the present invention, be not intended to limit the scope of the invention, it is every to utilize this hair
Equivalent structure or equivalent flow shift made by bright description is applied directly or indirectly in other relevant technology necks
Domain is included within the scope of the present invention.
Claims (5)
1. a kind of method of micro, quick measurement algae oil content, which is characterized in that described method includes following steps：
(1) algae solution for carrying out microdisk electrode is taken, the unit for calculating the number concentration C, C of microalgae cell in algae solution is cell/ml；
(2) wet algae samples of the 0.5-1.0g through desalination point is weighed, 50-100ml distilled water is added, is sufficiently mixed uniformly, then dilutes
At the algae solution of 1/2,1/3,1/4,1/5,1,/10 5 different concentration of algae；
(3) 0.4-0.8g fluorescent staining lipotropism fluorescent dyes powder is weighed, is dissolved in organic solvent, is placed 4-6 days at room temperature,
Period often shakes, and is filtered after allowing it fully to dissolve, is configured to mother liquor；
(4) above-mentioned each concentration of algae takes 2 test tubes respectively, each HCl that 5-10mL algae solutions and 5-10ml1mol/L is added, fully mixed
After conjunction, 0.1ul mother liquors are added in one branch pipe of each concentration of algae, and the organic solvent in step (3) is added in another branch pipe, as a contrast
Pipe, mixes well；
(5) excitation wavelength 400-600nm exciting lights are used, in fluorescence microscopy microscopic observation and photograph to record Unicell Oils and Fats accumulation
Situation of change；
(6) unicellular olesome number is recorded according to photo, and measures droplet diameter, each oil is calculated according to sphere volume formula
Drop volume, each cell grease content are the adduction of all blob volumes；
(7) according to the number concentration C of microalgae cell and individual cells fat content unit of account volume algae solution fat content：Vt=
CV, C are the number concentration of microalgae cell, and V is Unicell Oils and Fats volume, and Vt is unit volume fat content.
2. the method for micro, quick measurement algae oil content as described in claim 1, which is characterized in that before the step (2)
Further include：
(11) after the salinity of microalgae cell, centrifugation for several times are washed with retarder thinner, are removed in repetition, algae solution centrifugal concentrating.
3. the method for micro, quick measurement algae oil content as claimed in claim 2, which is characterized in that dilute in step (11)
It is distilled water, tap water, deionized water, ultra-clean water or mineral water to release solvent.
4. the method for micro, quick measurement algae oil content as described in claim 1, which is characterized in that the fluorescent staining parent
Lipid fluorescent dye is the Sudan three, Sudan IV or Sudan black B, and the organic solvent is 100-200ml70% ethyl alcohol.
5. the method for micro, quick measurement algae oil content as described in claim 1, which is characterized in that the step (5) is specific
For：After microalgae cell dyeing under 507nm excitation wavelengths, using laser scanning co-focusing microscope every 0.01-0.5 μm to micro-
Frustule tomoscan simultaneously photographs to record, and oil droplet 3D pictures is rebuild by 3D rendering composite software, using microphotograph software
Measure olesome diameter.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810165144.0A CN108535222A (en) | 2018-02-27 | 2018-02-27 | A kind of method of micro, quick measurement algae oil content |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810165144.0A CN108535222A (en) | 2018-02-27 | 2018-02-27 | A kind of method of micro, quick measurement algae oil content |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108535222A true CN108535222A (en) | 2018-09-14 |
Family
ID=63486201
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810165144.0A Pending CN108535222A (en) | 2018-02-27 | 2018-02-27 | A kind of method of micro, quick measurement algae oil content |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108535222A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113155800A (en) * | 2021-05-04 | 2021-07-23 | 浙江师范大学 | Method for observing and quantifying grease in grape seed material through laser confocal development |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102033059A (en) * | 2010-11-29 | 2011-04-27 | 哈尔滨工业大学 | Method for detecting microalgae oil |
| CN102297842A (en) * | 2010-06-22 | 2011-12-28 | 国家海洋局第三海洋研究所 | Method for rapidly testing lipid content of microalgae with sudan black B |
| CN102565012A (en) * | 2010-12-24 | 2012-07-11 | 中国水产科学研究院黄海水产研究所 | Method for detecting oil content of microalgae |
| CN103163113A (en) * | 2013-03-25 | 2013-06-19 | 哈尔滨工业大学 | Method for detecting content of microalgal oil by ultrasound-assisted fluorescent staining |
| CN103555587A (en) * | 2013-11-07 | 2014-02-05 | 三亚深海科学与工程研究所 | Method of screening high grease algae from natural water body |
-
2018
- 2018-02-27 CN CN201810165144.0A patent/CN108535222A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102297842A (en) * | 2010-06-22 | 2011-12-28 | 国家海洋局第三海洋研究所 | Method for rapidly testing lipid content of microalgae with sudan black B |
| CN102033059A (en) * | 2010-11-29 | 2011-04-27 | 哈尔滨工业大学 | Method for detecting microalgae oil |
| CN102565012A (en) * | 2010-12-24 | 2012-07-11 | 中国水产科学研究院黄海水产研究所 | Method for detecting oil content of microalgae |
| CN103163113A (en) * | 2013-03-25 | 2013-06-19 | 哈尔滨工业大学 | Method for detecting content of microalgal oil by ultrasound-assisted fluorescent staining |
| CN103555587A (en) * | 2013-11-07 | 2014-02-05 | 三亚深海科学与工程研究所 | Method of screening high grease algae from natural water body |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113155800A (en) * | 2021-05-04 | 2021-07-23 | 浙江师范大学 | Method for observing and quantifying grease in grape seed material through laser confocal development |
| CN113155800B (en) * | 2021-05-04 | 2023-11-07 | 浙江师范大学 | Method for observing and quantifying grease in grape seed material by using laser copolymerization Jiao Xianying |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102565012B (en) | Method for detecting oil content of microalgae | |
| CN106124475B (en) | A kind of trace pesticide residue Raman spectrum method for detecting surface reinforcement based on aptamer | |
| CN105675566A (en) | Method for quantitatively analyzing enrichment and distribution of micro-plastic in mammal body | |
| CN102288558B (en) | Method for quick microdetection of content of fat in microalga | |
| CN103555587A (en) | Method of screening high grease algae from natural water body | |
| CN102321729B (en) | Fluorescent microscopic counting method for detecting bacterial count in soil and sediment | |
| Cvjetinovic et al. | Photoacoustic and fluorescence lifetime imaging of diatoms | |
| CN108535222A (en) | A kind of method of micro, quick measurement algae oil content | |
| CN106404693B (en) | A method of two-dimension nano materials concentration in measurement two-dimensional material aaerosol solution | |
| CN102297842A (en) | Method for rapidly testing lipid content of microalgae with sudan black B | |
| CN108037104B (en) | Method for screening optimal oil-producing algae species based on fat-soluble quantum dots | |
| CN107238597A (en) | A kind of method of lignin micro Distribution rule in research wood cell wall | |
| CN105784573A (en) | Method for analyzing wheat root tip cell cycle through flow cytometry | |
| CN111829859A (en) | Efficient transparent dyeing and three-dimensional imaging method for poplar seeds | |
| CN109696393A (en) | A kind of method that determination of the environment tests polystyrene microsphere content under simulated conditions | |
| CN112940021A (en) | High-brightness multi-color washing-free fat-drop fluorescent probe | |
| CN105675375A (en) | Separating medium of inactivated nematode in dead masson pine wood and separation method and kit | |
| CN103018086A (en) | Pretreatment and measuring method for measuring botryococcus lipid content by using fluorescent spectrometry | |
| CN104345052B (en) | A kind of method that fluorescence negative staining detects living cells | |
| Ding et al. | An innovative miniaturized cell imaging system based on integrated coaxial dual optical path structure and microfluidic fixed frequency sample loading strategy | |
| CN112782137B (en) | Flow cytometer detection method for living plankton living cells with 10-50 mu m | |
| Deng et al. | Monitoring the life cycle stages of Haematococcus pluvialis by using FLIM | |
| CN114702688B (en) | Preparation method of centrifugal hydrogel liquid drops | |
| Bodénès et al. | A microfluidic device for the real-time characterization of lipid producing algae cell population submitted to a pulsed electric field | |
| CN110241047B (en) | Method for screening linear spirulina filaments by using triclosan |
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 | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: 3 / F, North Funuo health building, Yulu Avenue, Lankao County, Kaifeng City, Henan Province Applicant after: Funo health Co.,Ltd. Address before: 3 / F, North Funuo health building, Yulu Avenue, Lankao County, Kaifeng City, Henan Province Applicant before: Funo health (Lankao) Co.,Ltd. Address after: 3 / F, North Funuo health building, Yulu Avenue, Lankao County, Kaifeng City, Henan Province Applicant after: Funo health (Lankao) Co.,Ltd. Address before: Room 2511, Dart International Center, 238 Changgang Middle Road, Haizhu District, Guangzhou City, Guangdong Province Applicant before: GUANGZHOU FUNUO HEALTH TECHNOLOGY Co.,Ltd. |
|
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180914 |