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A flow cytometer based protocol for quantitative analysis of bloom-forming cyanobacteria (Microcystis) in lake sediments
Zhou, Quan1,2; Chen, Wei1; Zhang, Huiyong3; Peng, Liang1; Liu, Liming1; Han, Zhiguo3; Wan, Neng4; Li, Lin1; Song, Lirong1; Song, LR (reprint author), Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China.
2012
Source PublicationJOURNAL OF ENVIRONMENTAL SCIENCES-CHINA
ISSN1001-0742
Volume24Issue:9Pages:1709-1716
AbstractA quantitative protocol for the rapid analysis of Microcystis cells and colonies in lake sediment was developed using a modified flow cytometer, the CytoSense. For cell enumeration, diluted sediment samples containing Microcystis were processed with sonication to disintegrate colonies into single cells. An optimized procedure suggested that 5 mg dw (dry weight)/mL dilution combined with 200 W x 2 min sonication yielded the highest counting efficiency. Under the optimized determination conditions, the quantification limit of this protocol was 3.3x10(4) cells/g dw. For colony analysis, Microcystis were isolated from the sediment by filtration. Colony lengths measured by flow cytometry were similar to those measured by microscopy for the size range of one single cell to almost 400 mu m in length. Moreover, the relationship between colony size and cell number was determined for three Microcystis species, including Microcystis flos-aquae, M. aeruginosa and M. wessenbergii. Regression formulas were used to calculate the cell numbers in different-sized colonies. The developed protocol was applied to field sediment samples from Lake Taihu. The results indicated the potential and applicability of flow cytometry as a tool for the rapid analysis of benthic Microcystis. This study provided a new capability for the high frequency monitoring of benthic overwintering and population dynamics of this bloom-forming cyanobacterium.; A quantitative protocol for the rapid analysis of Microcystis cells and colonies in lake sediment was developed using a modified flow cytometer, the CytoSense. For cell enumeration, diluted sediment samples containing Microcystis were processed with sonication to disintegrate colonies into single cells. An optimized procedure suggested that 5 mg dw (dry weight)/mL dilution combined with 200 W x 2 min sonication yielded the highest counting efficiency. Under the optimized determination conditions, the quantification limit of this protocol was 3.3x10(4) cells/g dw. For colony analysis, Microcystis were isolated from the sediment by filtration. Colony lengths measured by flow cytometry were similar to those measured by microscopy for the size range of one single cell to almost 400 mu m in length. Moreover, the relationship between colony size and cell number was determined for three Microcystis species, including Microcystis flos-aquae, M. aeruginosa and M. wessenbergii. Regression formulas were used to calculate the cell numbers in different-sized colonies. The developed protocol was applied to field sediment samples from Lake Taihu. The results indicated the potential and applicability of flow cytometry as a tool for the rapid analysis of benthic Microcystis. This study provided a new capability for the high frequency monitoring of benthic overwintering and population dynamics of this bloom-forming cyanobacterium.
SubtypeArticle
KeywordFlow Cytometry Benthic Microcystis Cell Enumeration Colony Size Measurement
Department[Zhou, Quan; Chen, Wei; Peng, Liang; Liu, Liming; Li, Lin; Song, Lirong] Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China; [Zhou, Quan] Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China; [Zhang, Huiyong; Han, Zhiguo] Zealquest Sci Technol Co Ltd, Zealquest Lab Ecol Res, Shanghai 200333, Peoples R China; [Wan, Neng] Changshu Inst Technol, Changshu 215500, Peoples R China
DOI10.1016/S1001-0742(11)60993-5
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Funding OrganizationNational Basic Research Program (973) of China [2008CB418006]; National Special Program of Water Environment [2009ZX07106-001-002]; National Natural Science Foundation of China [31070355]; National Major Science and Technology Program for Water Pollution Control and Treatment [2009ZX07101-013] ; National Basic Research Program (973) of China [2008CB418006]; National Special Program of Water Environment [2009ZX07106-001-002]; National Natural Science Foundation of China [31070355]; National Major Science and Technology Program for Water Pollution Control and Treatment [2009ZX07101-013] ; National Basic Research Program (973) of China [2008CB418006]; National Special Program of Water Environment [2009ZX07106-001-002]; National Natural Science Foundation of China [31070355]; National Major Science and Technology Program for Water Pollution Control and Treatment [2009ZX07101-013] ; National Basic Research Program (973) of China [2008CB418006]; National Special Program of Water Environment [2009ZX07106-001-002]; National Natural Science Foundation of China [31070355]; National Major Science and Technology Program for Water Pollution Control and Treatment [2009ZX07101-013]
Indexed BySCI
Language英语
WOS Research AreaEnvironmental Sciences & Ecology
WOS SubjectEnvironmental Sciences
WOS IDWOS:000309371600022
WOS KeywordAERUGINOSA KUTZ ; EUTROPHIC LAKE ; MEILIANG BAY ; PHYTOPLANKTON ; CYANOPHYCEAE ; RECRUITMENT ; SHALLOW ; MARINE ; WATER ; CYTOBUOY
Funding OrganizationNational Basic Research Program (973) of China [2008CB418006]; National Special Program of Water Environment [2009ZX07106-001-002]; National Natural Science Foundation of China [31070355]; National Major Science and Technology Program for Water Pollution Control and Treatment [2009ZX07101-013] ; National Basic Research Program (973) of China [2008CB418006]; National Special Program of Water Environment [2009ZX07106-001-002]; National Natural Science Foundation of China [31070355]; National Major Science and Technology Program for Water Pollution Control and Treatment [2009ZX07101-013] ; National Basic Research Program (973) of China [2008CB418006]; National Special Program of Water Environment [2009ZX07106-001-002]; National Natural Science Foundation of China [31070355]; National Major Science and Technology Program for Water Pollution Control and Treatment [2009ZX07101-013] ; National Basic Research Program (973) of China [2008CB418006]; National Special Program of Water Environment [2009ZX07106-001-002]; National Natural Science Foundation of China [31070355]; National Major Science and Technology Program for Water Pollution Control and Treatment [2009ZX07101-013]
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/17282
Collection藻类生物学及应用研究中心_期刊论文
Corresponding AuthorSong, LR (reprint author), Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China.
Affiliation1.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
3.Zealquest Sci Technol Co Ltd, Zealquest Lab Ecol Res, Shanghai 200333, Peoples R China
4.Changshu Inst Technol, Changshu 215500, Peoples R China
Recommended Citation
GB/T 7714
Zhou, Quan,Chen, Wei,Zhang, Huiyong,et al. A flow cytometer based protocol for quantitative analysis of bloom-forming cyanobacteria (Microcystis) in lake sediments[J]. JOURNAL OF ENVIRONMENTAL SCIENCES-CHINA,2012,24(9):1709-1716.
APA Zhou, Quan.,Chen, Wei.,Zhang, Huiyong.,Peng, Liang.,Liu, Liming.,...&Song, LR .(2012).A flow cytometer based protocol for quantitative analysis of bloom-forming cyanobacteria (Microcystis) in lake sediments.JOURNAL OF ENVIRONMENTAL SCIENCES-CHINA,24(9),1709-1716.
MLA Zhou, Quan,et al."A flow cytometer based protocol for quantitative analysis of bloom-forming cyanobacteria (Microcystis) in lake sediments".JOURNAL OF ENVIRONMENTAL SCIENCES-CHINA 24.9(2012):1709-1716.
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