Publication
Development of a short-term assay based on the evaluation of the plasma membrane integrity of the alga Pseudokirchneriella subcapitata
| dc.contributor.author | Machado, Manuela D. | |
| dc.contributor.author | Soares, Eduardo V. | |
| dc.date.accessioned | 2017-09-14T13:42:10Z | |
| dc.date.available | 2017-09-14T13:42:10Z | |
| dc.date.issued | 2012 | |
| dc.description.abstract | Membrane integrity has been used as a criterion for the definition of cell viability. In the present work, staining conditions (time and dye concentration) for the evaluation of membrane integrity in a fluorescence microplate reader, using the membrane-impermeant nucleic-acid dye SYTOX Green, were optimized. Incubating Pseudokirchneriella subcapitata algal cells with 0.5 μmol/l SYTOX Green for 40 min allowed a clear discrimination between live (intact plasma membrane) and dead cells (with compromised plasma membrane). Algal cell suspensions, labelled with SYTOX Green, exhibited a green fluorescence proportional to the fraction of the cells with a permeabilized plasma membrane. The optimized staining conditions were used to assess the toxicity of 1-pentanol on P. subcapitata in a short-term exposure (6 h) assay. The loss of membrane integrity in the cell population increased with the concentration of 1-pentanol. The 6-h EC10 and EC50 values were 7,617 mg/l 1-pentanol (95 % confidence limits 4,670–9,327) and 12,818 mg/l 1-pentanol (95 % confidence limits 10,929–15,183), respectively. The developed microplate-based short-term assay can be useful in the highthroughput screening of toxics or environmental samples using the alga P. subcapitata. | pt_PT |
| dc.description.version | info:eu-repo/semantics/publishedVersion | pt_PT |
| dc.identifier.doi | 10.1007/s00253-012-4185-y | pt_PT |
| dc.identifier.uri | http://hdl.handle.net/10400.22/10262 | |
| dc.language.iso | eng | pt_PT |
| dc.publisher | Springer Verlag | pt_PT |
| dc.relation.ispartofseries | Applied Microbiology and Biotechnology;Vol. 95, Issue 4 | |
| dc.subject | Membrane integrity | pt_PT |
| dc.subject | Microplate assay | pt_PT |
| dc.subject | Selenastrum capricornutum | pt_PT |
| dc.subject | 1-Pentanol | pt_PT |
| dc.subject | SYTOX Green | pt_PT |
| dc.subject | Acute toxicity | pt_PT |
| dc.title | Development of a short-term assay based on the evaluation of the plasma membrane integrity of the alga Pseudokirchneriella subcapitata | pt_PT |
| dc.type | journal article | |
| dspace.entity.type | Publication | |
| oaire.citation.title | Applied Microbiology and Biotechnology | pt_PT |
| person.familyName | Soares | |
| person.givenName | Eduardo | |
| person.identifier | 93637 | |
| person.identifier.ciencia-id | D612-0D0E-4984 | |
| person.identifier.orcid | 0000-0003-2280-5291 | |
| person.identifier.rid | O-5908-2015 | |
| person.identifier.scopus-author-id | 7102060173 | |
| rcaap.rights | openAccess | pt_PT |
| rcaap.type | article | pt_PT |
| relation.isAuthorOfPublication | b3ebdee1-84fa-4183-a1d8-c3cdcaaef219 | |
| relation.isAuthorOfPublication.latestForDiscovery | b3ebdee1-84fa-4183-a1d8-c3cdcaaef219 |