Evaluation of confocal laser scanning microscopy for enumeration of virus-like particles in aquatic systems
Abundances of virus-like particles (VLPs, mostly bacteriophages) are high in aquatic environments; therefore, techniques for precise enumeration are essential in ecological monitoring. VLPs were determined after staining with SYBR Gold by conventional epifluorescence microscopy and compared to enume...
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| Veröffentlicht in: | Environmental monitoring and assessment 2013-07, Vol.185 (7), p.5411-5418 |
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| description | Abundances of virus-like particles (VLPs, mostly bacteriophages) are high in aquatic environments; therefore, techniques for precise enumeration are essential in ecological monitoring. VLPs were determined after staining with SYBR Gold by conventional epifluorescence microscopy and compared to enumerations performed by confocal laser scanning microscopy (CLSM). In order to assess the potential of CLSM for viral direct counts (VDCs), we processed samples from different freshwater and marine systems. Optical sectioning by CLSM and production of an overlay picture of multiple scans enables the often uneven whole investigated filter area to be brought to the plane of focus. This allows for subsequent image analysis of digitally created high-quality images. Another advantage using the CLSM was that the short spot excitation of the stain via laser beam minimized fading of the stain. The VDC results show that there is no significant difference between the two methods. Regarding the known difficulties of viral abundance estimates on particulate material, CLSM was further applied to enumerate VLPs on a small set of marine transparent exopolymeric particles sampled from the Atlantic Ocean. Our data suggest that CLSM is a useful tool to count viruses in water samples as well as attached to certain types of aquatic aggregates. |
| doi_str_mv | 10.1007/s10661-012-2955-8 |
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VLPs were determined after staining with SYBR Gold by conventional epifluorescence microscopy and compared to enumerations performed by confocal laser scanning microscopy (CLSM). In order to assess the potential of CLSM for viral direct counts (VDCs), we processed samples from different freshwater and marine systems. Optical sectioning by CLSM and production of an overlay picture of multiple scans enables the often uneven whole investigated filter area to be brought to the plane of focus. This allows for subsequent image analysis of digitally created high-quality images. Another advantage using the CLSM was that the short spot excitation of the stain via laser beam minimized fading of the stain. The VDC results show that there is no significant difference between the two methods. Regarding the known difficulties of viral abundance estimates on particulate material, CLSM was further applied to enumerate VLPs on a small set of marine transparent exopolymeric particles sampled from the Atlantic Ocean. Our data suggest that CLSM is a useful tool to count viruses in water samples as well as attached to certain types of aquatic aggregates.</description><identifier>ISSN: 0167-6369</identifier><identifier>EISSN: 1573-2959</identifier><identifier>DOI: 10.1007/s10661-012-2955-8</identifier><identifier>PMID: 23108709</identifier><identifier>CODEN: EMASDH</identifier><language>eng</language><publisher>Dordrecht: Springer Netherlands</publisher><subject>Aggregates ; Analysis ; Animal, plant and microbial ecology ; Applied ecology ; Aquatic ecosystems ; Aquatic environment ; Atlantic Ocean ; Atmospheric Protection/Air Quality Control/Air Pollution ; Bacteria ; bacteriophages ; Biofilms ; Biological and medical sciences ; confocal laser scanning microscopy ; Confocal microscopy ; Conservation, protection and management of environment and wildlife ; Earth and Environmental Science ; Ecological monitoring ; Ecology ; Ecotoxicology ; Enumeration ; Environment ; Environmental Management ; Environmental Monitoring ; Environmental Monitoring - methods ; filters ; Fresh Water ; Fresh Water - virology ; freshwater ; Fundamental and applied biological sciences. Psychology ; growth & development ; image analysis ; Image processing ; isolation & purification ; Lasers ; Marine systems ; methods ; Microscopy ; Microscopy, Confocal ; monitoring ; Monitoring/Environmental Analysis ; Particulate matter ; Phages ; Seawater ; Seawater - virology ; Sectioning ; Stains & staining ; Studies ; Virion ; Virion - growth & development ; Virion - isolation & purification ; virology ; Virus-like particles ; Viruses ; Water analysis ; Water sampling</subject><ispartof>Environmental monitoring and assessment, 2013-07, Vol.185 (7), p.5411-5418</ispartof><rights>Springer Science+Business Media Dordrecht 2012</rights><rights>2014 INIST-CNRS</rights><rights>Springer Science+Business Media Dordrecht 2013</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c566t-c83e00ff4a768bbd7c1ec3d6f85e4bfb2a99bedd707fff6a4438eadc916c42543</citedby><cites>FETCH-LOGICAL-c566t-c83e00ff4a768bbd7c1ec3d6f85e4bfb2a99bedd707fff6a4438eadc916c42543</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s10661-012-2955-8$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s10661-012-2955-8$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>230,314,777,781,882,27905,27906,41469,42538,51300</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=27605709$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23108709$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Peduzzi, Peter</creatorcontrib><creatorcontrib>Agis, Martin</creatorcontrib><creatorcontrib>Luef, Birgit</creatorcontrib><title>Evaluation of confocal laser scanning microscopy for enumeration of virus-like particles in aquatic systems</title><title>Environmental monitoring and assessment</title><addtitle>Environ Monit Assess</addtitle><addtitle>Environ Monit Assess</addtitle><description>Abundances of virus-like particles (VLPs, mostly bacteriophages) are high in aquatic environments; therefore, techniques for precise enumeration are essential in ecological monitoring. VLPs were determined after staining with SYBR Gold by conventional epifluorescence microscopy and compared to enumerations performed by confocal laser scanning microscopy (CLSM). In order to assess the potential of CLSM for viral direct counts (VDCs), we processed samples from different freshwater and marine systems. Optical sectioning by CLSM and production of an overlay picture of multiple scans enables the often uneven whole investigated filter area to be brought to the plane of focus. This allows for subsequent image analysis of digitally created high-quality images. Another advantage using the CLSM was that the short spot excitation of the stain via laser beam minimized fading of the stain. The VDC results show that there is no significant difference between the two methods. Regarding the known difficulties of viral abundance estimates on particulate material, CLSM was further applied to enumerate VLPs on a small set of marine transparent exopolymeric particles sampled from the Atlantic Ocean. Our data suggest that CLSM is a useful tool to count viruses in water samples as well as attached to certain types of aquatic aggregates.</description><subject>Aggregates</subject><subject>Analysis</subject><subject>Animal, plant and microbial ecology</subject><subject>Applied ecology</subject><subject>Aquatic ecosystems</subject><subject>Aquatic environment</subject><subject>Atlantic Ocean</subject><subject>Atmospheric Protection/Air Quality Control/Air Pollution</subject><subject>Bacteria</subject><subject>bacteriophages</subject><subject>Biofilms</subject><subject>Biological and medical sciences</subject><subject>confocal laser scanning microscopy</subject><subject>Confocal microscopy</subject><subject>Conservation, protection and management of environment and wildlife</subject><subject>Earth and Environmental Science</subject><subject>Ecological monitoring</subject><subject>Ecology</subject><subject>Ecotoxicology</subject><subject>Enumeration</subject><subject>Environment</subject><subject>Environmental Management</subject><subject>Environmental Monitoring</subject><subject>Environmental Monitoring - methods</subject><subject>filters</subject><subject>Fresh Water</subject><subject>Fresh Water - virology</subject><subject>freshwater</subject><subject>Fundamental and applied biological sciences. 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VLPs were determined after staining with SYBR Gold by conventional epifluorescence microscopy and compared to enumerations performed by confocal laser scanning microscopy (CLSM). In order to assess the potential of CLSM for viral direct counts (VDCs), we processed samples from different freshwater and marine systems. Optical sectioning by CLSM and production of an overlay picture of multiple scans enables the often uneven whole investigated filter area to be brought to the plane of focus. This allows for subsequent image analysis of digitally created high-quality images. Another advantage using the CLSM was that the short spot excitation of the stain via laser beam minimized fading of the stain. The VDC results show that there is no significant difference between the two methods. Regarding the known difficulties of viral abundance estimates on particulate material, CLSM was further applied to enumerate VLPs on a small set of marine transparent exopolymeric particles sampled from the Atlantic Ocean. Our data suggest that CLSM is a useful tool to count viruses in water samples as well as attached to certain types of aquatic aggregates.</abstract><cop>Dordrecht</cop><pub>Springer Netherlands</pub><pmid>23108709</pmid><doi>10.1007/s10661-012-2955-8</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Environmental monitoring and assessment, 2013-07, Vol.185 (7), p.5411-5418 |
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| language | eng |
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| source | MEDLINE; Springer Nature - Complete Springer Journals |
| subjects | Aggregates Analysis Animal, plant and microbial ecology Applied ecology Aquatic ecosystems Aquatic environment Atlantic Ocean Atmospheric Protection/Air Quality Control/Air Pollution Bacteria bacteriophages Biofilms Biological and medical sciences confocal laser scanning microscopy Confocal microscopy Conservation, protection and management of environment and wildlife Earth and Environmental Science Ecological monitoring Ecology Ecotoxicology Enumeration Environment Environmental Management Environmental Monitoring Environmental Monitoring - methods filters Fresh Water Fresh Water - virology freshwater Fundamental and applied biological sciences. Psychology growth & development image analysis Image processing isolation & purification Lasers Marine systems methods Microscopy Microscopy, Confocal monitoring Monitoring/Environmental Analysis Particulate matter Phages Seawater Seawater - virology Sectioning Stains & staining Studies Virion Virion - growth & development Virion - isolation & purification virology Virus-like particles Viruses Water analysis Water sampling |
| title | Evaluation of confocal laser scanning microscopy for enumeration of virus-like particles in aquatic systems |
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