CN103484523A - Method for counting microcystis population cells - Google Patents
Method for counting microcystis population cells Download PDFInfo
- Publication number
- CN103484523A CN103484523A CN201310425334.9A CN201310425334A CN103484523A CN 103484523 A CN103484523 A CN 103484523A CN 201310425334 A CN201310425334 A CN 201310425334A CN 103484523 A CN103484523 A CN 103484523A
- Authority
- CN
- China
- Prior art keywords
- microcystis
- microcystis aeruginosa
- sample
- counting
- described step
- 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.)
- Granted
Links
Images
Landscapes
- Separation Of Suspended Particles By Flocculating Agents (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The method discloses a method for counting microcystis population cells. First, a water sample of 1L is collected from natural water at the outburst acme of microcystis water blooms, then, the collected water sample is filtered through a microfiltration membrane with the pore diameter of about 0.45 microns, next, microcystis population samples attached to the microfiltration membrane after the filtration are suspended in a 100mL beaker containing 10mL of 10<-3>mol/L EDTA solution, after 100 small glass beads are added into the beaker, the solution in the beaker is stirred for 100min magnetically at 1600rm/min, totally dispersed single-cell microcystis samples are obtained and undamaged, and finally the microcystis cells are counted with a blood counting chamber. The method for counting microcystis population cells has the advantages of simplifying experimental steps, and being short in consumed time and capable of being used for accurately counting the microcystis population cells.
Description
Technical field
The invention belongs to the water pollution control field, particularly relate to the somatic method of counting of a kind of Microcystis aeruginosa group.
Background technology
Microcystis aeruginosa (Microcystis sp.) is the bloom blue algae kind of blazoning property of the whole world, is also modal blue-green alga bloom main composition kind in China's eutrophication water.Microcystis aeruginosa usually exists with unicellular or two cells form under the laboratory culture condition, and is to exist with a large amount of macroscopic group forms in natural water in the wild.The now relevant somatic method of counting of field Microcystis aeruginosa group has whirlpool concussion method, boiling method, titanium dioxide or ultraviolet treatment, yet capsule algae colony cell can't be separated into unicellular fully under the effect of whirlpool concussion method, and Microcystis aeruginosa colony cell all has breakage in various degree under boiling method, titanium dioxide or ultraviolet treatment effect, thereby above-mentioned method of counting has all reduced the Cytometric accuracy of Microcystis aeruginosa colony to some extent.
Summary of the invention
Purpose of the present invention just is to provide a kind of Microcystis aeruginosa group somatic method of counting, often break out the monitoring of the water body Microcystis aeruginosa density of blue-green alga bloom for various poisons in freshwaters such as artificial water application's system, lake, reservoir and ponds, for the routine monitoring of natural water microcystis number and early warning and the control of microcystis waterbloom.
In order to solve the problems of the technologies described above, the technical solution used in the present invention is: the somatic method of counting of a kind of Microcystis aeruginosa group comprises the steps:
(1) quantitatively take the water sample of the natural water of microcystis waterbloom outburst Sheng phase;
(2) the water sample via hole diameter collected is not more than to the screen filtration of 2 μ m, the Microcystis aeruginosa sample is attached on screen cloth;
(3) filtering the Microcystis aeruginosa colony sample of postadhesion on screen cloth, again to be suspended in concentration be 10
-3in the beaker of mol/L EDTA solution;
(4) after adding little granulated glass sphere to above-mentioned beaker, magnetic agitation;
(5) draw the unicellular sample of Microcystis aeruginosa that micro-Microscopic observation spreads out fully, adopt blood counting chamber counting microcystis number.
In described step (2), the millipore filtration that described screen cloth is aperture 0.45 μ m or aperture are not more than the bolting silk of 2 μ m.
In described step (3), utilize liquid-transfering gun to draw EDTA solution and repeatedly rinse screen cloth, until Microcystis aeruginosa colony elutes from screen cloth fully.
In described step (4), in the magnetic agitation process, every mistake is checked Microcystis aeruginosa colony cell dispersion process in 5 minutes under the microscope, until Microcystis aeruginosa colony cell is separated into unicellular fully.
In described step (4), turn/min of magnetic agitation speed 1000-2000, magnetic agitation time 90-110min.
In described step (5), before drawing the unicellular sample of Microcystis aeruginosa, draw again after mixing, and blood counting chamber is counted to the microcystis quantity that the microcystis number is converted into every premium on currency sample.
The invention has the beneficial effects as follows: capsule algae colony cell under the effect of whirlpool concussion method can be overcome and the defect that Microcystis aeruginosa colony cell under unicellular and boiling method, titanium dioxide or ultraviolet treatment effect all has breakage in various degree can't be separated into fully, but the somatic number of Microcystis aeruginosa group in the accurate counting natural water.Experimental procedure is simple, consuming time shorter, and can accurately improve the somatic number of counting Microcystis aeruginosa group.Be widely used in the prevention and control of the various poisons in freshwater microcystis waterblooms such as artificial water application's system, lake, reservoir and pond.
The accompanying drawing explanation
Fig. 1 is the variation of Microcystis aeruginosa colony form of the first embodiment under the inventive method effect.
Fig. 2 is the variation of Microcystis aeruginosa colony form of the second embodiment under the inventive method effect.
Embodiment
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail:
The somatic method of counting of Microcystis aeruginosa group of the present invention, comprise the steps:
(1) quantitatively take the water sample of the natural water of microcystis waterbloom outburst Sheng phase;
(2) the water sample via hole diameter collected is not more than to the screen filtration of 2 μ m, the Microcystis aeruginosa sample is attached on screen cloth;
(3) filtering the Microcystis aeruginosa colony sample of postadhesion on screen cloth, again to be suspended in concentration be 10
-3in the beaker of mol/L EDTA solution;
(4) after adding little granulated glass sphere to above-mentioned beaker, magnetic agitation;
(5) draw the unicellular sample of Microcystis aeruginosa that micro-Microscopic observation spreads out fully, adopt blood counting chamber counting microcystis number.
In described step (2), the millipore filtration that described screen cloth is aperture 0.45 μ m or aperture are not more than the bolting silk of 2 μ m.
In described step (3), utilize liquid-transfering gun to draw EDTA solution and repeatedly rinse screen cloth, until Microcystis aeruginosa colony elutes from screen cloth fully.
In described step (4), in the magnetic agitation process, every mistake is checked Microcystis aeruginosa colony cell dispersion process in 5 minutes under the microscope, until Microcystis aeruginosa colony cell is separated into unicellular fully.
In described step (4), turn/min of magnetic agitation speed 1000-2000, magnetic agitation time 90-110min.
In described step (5), before drawing the unicellular sample of Microcystis aeruginosa, draw again after mixing, and blood counting chamber is counted to the microcystis quantity that the microcystis number is converted into every premium on currency sample.
High speed magnetic stirring, in conjunction with the effect of granulated glass sphere, can elute bacterium and fungal cell's exocellular polysaccharide and do not destroy bacterium or fungal cell's integrity under the cooperation of EDTA solution.
Embodiment 1 the inventive method is applied to the example of natural water microcystis measuring density
Microcystis number while using the present invention to count Tianjin, Tianjin river valley eight li platform section outburst microcystis waterblooms on July 3rd, 2013:
(1) utilize the water sampler of 1L to take river, Tianjin microcystis waterbloom to break out the water sample 1L of the natural water of Sheng phase; (2) Tianjin river sample via hole diameter collected is about to the filtering with microporous membrane of 0.45 μ m, the Microcystis aeruginosa sample is attached on millipore filtration;
(3) will filter the Microcystis aeruginosa colony sample of postadhesion on millipore filtration again is suspended in and contains 10mL10
-3in the 100mL beaker of mol/L EDTA solution;
(4) after adding 100 little granulated glass spherees to above-mentioned 100mL beaker, 1600rm/min magnetic agitation 90-110min; (see figure 1)
(5) draw the unicellular sample of Microcystis aeruginosa that micro-Microscopic observation spreads out fully, adopt hemocytometer
Number plate counting microcystis number.
In described step (3), utilize liquid-transfering gun to draw EDTA solution and repeatedly rinse screen cloth, until Microcystis aeruginosa colony elutes from screen cloth fully.
In described step (4), in the magnetic agitation process, every mistake is checked Microcystis aeruginosa colony cell dispersion process in 5 minutes under the microscope, until Microcystis aeruginosa colony cell is separated into unicellular fully.
In described step (5), before drawing the unicellular sample of Microcystis aeruginosa, after mixing, draw again.
Microcystis when the enforcement of method of the present invention obtains the eight li platform section outburst microcystis waterblooms in Jin river July 3 in 2013 (containing verdigris Microcystis aeruginosa M.aeruginosa and fish evil Microcystis aeruginosa M.ichthyoblabe) density is 2.42 * 10
8ind/L.
Embodiment 2 the inventive method are applied to the example of aquaculture water microcystis measuring density
On July 5th, 2013, microcystis (containing verdigris Microcystis aeruginosa M.aeruginosa and the fish evil Microcystis aeruginosa M.ichthyoblabe) density that records No. 3 pool cultivated water bodys of carp seed multiplication farm according to the inventive method was 6.42 * 10 at TanJin Agricultural College carp seed multiplication farm
7ind/L.
Method, with embodiment 1, just changes the millipore filtration of aperture 0.45 μ m into bolting silk that aperture is not more than 2 μ m, the results are shown in Figure 2.
Above-described embodiment is only for illustrating technological thought of the present invention and characteristics, its purpose is to make those skilled in the art can understand content of the present invention and implement according to this, can not only with the present embodiment, limit the scope of the claims of the present invention, be equal variation or the modification that all disclosed spirit is done, still drop in the scope of the claims of the present invention.
Claims (6)
1. the somatic method of counting of Microcystis aeruginosa group, is characterized in that, comprises the steps:
(1) quantitatively take the water sample of the natural water of microcystis waterbloom outburst Sheng phase;
(2) the water sample via hole diameter collected is not more than to the screen filtration of 2 μ m, the Microcystis aeruginosa sample is attached on screen cloth;
(3) filtering the Microcystis aeruginosa colony sample of postadhesion on screen cloth, again to be suspended in concentration be 10
-3in the beaker of mol/L EDTA solution;
(4) after adding little granulated glass sphere to above-mentioned beaker, magnetic agitation;
(5) draw the unicellular sample of Microcystis aeruginosa that micro-Microscopic observation spreads out fully, adopt blood counting chamber counting microcystis number.
2. the Cytometric method of Microcystis aeruginosa colony according to claim 1, is characterized in that, in described step (2), the millipore filtration that described screen cloth is aperture 0.45 μ m or aperture are not more than the bolting silk of 2 μ m.
3. the Cytometric method of Microcystis aeruginosa colony according to claim 1, is characterized in that, in described step (3), utilizes liquid-transfering gun to draw EDTA solution and repeatedly rinse screen cloth, until Microcystis aeruginosa colony elutes from screen cloth fully.
4. the Cytometric method of Microcystis aeruginosa colony according to claim 1, it is characterized in that, in described step (4), in the magnetic agitation process, every mistake is checked Microcystis aeruginosa colony cell dispersion process in 5 minutes under the microscope, until Microcystis aeruginosa colony cell is separated into unicellular fully.
5. the Cytometric method of Microcystis aeruginosa colony according to claim 4, is characterized in that, in described step (4), and turn/min of magnetic agitation speed 1000-2000, magnetic agitation time 90-110min.
6. the Cytometric method of Microcystis aeruginosa colony according to claim 1, it is characterized in that, in described step (5), before drawing the unicellular sample of Microcystis aeruginosa, draw again after mixing, and blood counting chamber is counted to the microcystis quantity that the microcystis number is converted into every premium on currency sample.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310425334.9A CN103484523B (en) | 2013-09-18 | 2013-09-18 | The somatic method of counting of a kind of Microcystis aeruginosa group |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310425334.9A CN103484523B (en) | 2013-09-18 | 2013-09-18 | The somatic method of counting of a kind of Microcystis aeruginosa group |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103484523A true CN103484523A (en) | 2014-01-01 |
| CN103484523B CN103484523B (en) | 2016-01-27 |
Family
ID=49825115
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310425334.9A Expired - Fee Related CN103484523B (en) | 2013-09-18 | 2013-09-18 | The somatic method of counting of a kind of Microcystis aeruginosa group |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103484523B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106018245A (en) * | 2016-05-20 | 2016-10-12 | 河海大学 | Method for enriching and detecting microcystis single cell and colony biomass |
| CN111562210A (en) * | 2020-06-16 | 2020-08-21 | 北京挑战农业科技有限公司 | Method for detecting viable count in pre-coated forage microecological preparation product |
| CN112881632A (en) * | 2021-01-20 | 2021-06-01 | 深圳市水文水质中心 | Method and device for counting algae in water sample |
-
2013
- 2013-09-18 CN CN201310425334.9A patent/CN103484523B/en not_active Expired - Fee Related
Non-Patent Citations (7)
| Title |
|---|
| BI XIANG-DONG ET AL.: "Effects of lead(II) on the extracellular polysaccharide(EPS) production and colony formation of cultured Microcystis aeruginosa", 《WATER SCIENCE & TECHNOLOGY》 * |
| 张祖建等: "水稻胚乳细胞计数方法研究", 《江苏农学院学报》 * |
| 杨波等: "蓝藻伪空胞的测定及其前处理方法研究", 《安徽农业科学》 * |
| 章文军等: "膜过滤细胞直接计数新方法", 《生物学杂志》 * |
| 舒天阁等: "低功率超声波去除铜绿微囊藻技术", 《华侨大学学报(自然科学版)》 * |
| 赵洋甬等: "微囊藻细胞计数法研究", 《福建分析测试》 * |
| 龚守良: "《放射医学实验教程》", 28 February 2009, 原子能出版社 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106018245A (en) * | 2016-05-20 | 2016-10-12 | 河海大学 | Method for enriching and detecting microcystis single cell and colony biomass |
| CN111562210A (en) * | 2020-06-16 | 2020-08-21 | 北京挑战农业科技有限公司 | Method for detecting viable count in pre-coated forage microecological preparation product |
| CN111562210B (en) * | 2020-06-16 | 2023-01-03 | 北京挑战农业科技有限公司 | Method for detecting number of viable bacteria in pre-coated feed microecological preparation product |
| CN112881632A (en) * | 2021-01-20 | 2021-06-01 | 深圳市水文水质中心 | Method and device for counting algae in water sample |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103484523B (en) | 2016-01-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Verlecar et al. | Phytoplankton identification manual | |
| CN103484523B (en) | The somatic method of counting of a kind of Microcystis aeruginosa group | |
| CN104792806A (en) | Plant pollen tube cell calcium ion ultrastructure positioning method | |
| CN103553227A (en) | Ultrafiltration water purifier | |
| Zhang et al. | Harvesting of Isochrysis zhanjiangensis using ultrafiltration: Changes in the contribution ratios of cells and algogenic organic matter to membrane fouling under different cross-flow velocities | |
| CN106018245A (en) | Method for enriching and detecting microcystis single cell and colony biomass | |
| Ma et al. | Effect of filter-feeding fish silver carp on phytoplankton species and size distribution in surface water: A field study in water works | |
| CN105950472A (en) | High-throughput microorganism isolation culture method | |
| CN107964530B (en) | Duckweed protoplast preparation method | |
| CN106035395B (en) | The preparation method of algicide base-material, algicide and its preparation method and application | |
| CN106442488B (en) | method for detecting living organisms with size of more than 50 mu m in ballast water | |
| WO2023074716A1 (en) | Method and system for collecting microplastic from water | |
| Diao et al. | Evaluation of morphological variation and biomass growth of Nostoc commune under laboratory conditions | |
| CN102620974A (en) | Method for enriching and purifying algae cells from suspended solids of natural water | |
| CN104726337A (en) | Method for enriching and purifying cryptosporidium and giardia in alga-containing water, and method for determining content of cryptosporidium and giardia | |
| CN204422311U (en) | A kind of marine bacterioplankton and viral sample fast separation device | |
| CN208902490U (en) | A kind of heavy metal in sea water pretreatment unit | |
| CN107964529A (en) | Horehound method for preparing protoplast | |
| CN207488048U (en) | Flow cytometer sample preparation cell filter | |
| CN106044878B (en) | A kind of blue algae treatment agent and its application method and application | |
| CN206089224U (en) | Water purifying device | |
| CN114292753B (en) | Method for separating cells and discharging nanoparticles from cells | |
| Falkner et al. | Phosphate uptake by blue green algae in vitro and in a lake during an algal bloom: Useful application of a force-flow relationship | |
| Heidman et al. | Selective feeding on nutrient-rich particles by gizzard shad Dorosoma cepedianum does not involve mechanical sorting | |
| CN103940656B (en) | A kind of Usnea Fructus Ananadis comosi nucleus suspension preparation method |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160127 Termination date: 20200918 |