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dc.contributor.author
Salzman, Valentina
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dc.contributor.author
Porro, Valentina
dc.contributor.author
Bollati Fogolín, Mariela
dc.contributor.author
Aguilar, Pablo Sebastián
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dc.date.available
2018-06-19T18:03:37Z
dc.date.issued
2015-09
dc.identifier.citation
Salzman, Valentina; Porro, Valentina; Bollati Fogolín, Mariela; Aguilar, Pablo Sebastián; Quantitation of yeast cell-cell fusion using multicolor flow cytometry; Wiley-liss, Div John Wiley & Sons Inc; Cytometry Part A; 87; 9; 9-2015; 843-854
dc.identifier.issn
1552-4922
dc.identifier.uri
http://hdl.handle.net/11336/49348
dc.description.abstract
Mating of haploid Saccharomyces cerevisiae cells of opposite sex provides a powerful model system to study the cell-cell fusion. However, a rapid and standardized method is much needed for quantitative assessment of fusion efficiency. The gold standard method relies on counting mating pairs in fluorescence microscopy images. This current method is limited by expectancy bias and it is time consuming, restricting the number of both cell-cell fusion events and strains that can be analyzed at once. Automatic approaches present a solution to these limitations. Here, we describe a novel flow cytometric approach that is able to quickly both identify mating pairs within a mixture of gametes and quantify cell fusion efficiency. This method is based on staining the cell wall of yeast populations with different Concanavalin A-fluorophore conjugates. The mating subpopulation is identified as the two-colored events set and fused and unfused mating pairs are subsequently discriminated by green fluorescent protein bimolecular complementation. A series of experiments was conducted to validate a simple and reliable protocol. Mating efficiency in each sample was determined by flow cytometry and compared with the one obtained with the current gold standard technique. The results show that mating pair counts using both methods produce indistinguishable outcomes and that the flow cytometry-based method provides quantitative relevant information in a short time, making possible to quickly analyze many different cell populations. In conclusion, our data show multicolor flow cytometry-based fusion quantitation to be a fast, robust, and reliable method to quantify the cell-cell fusion in yeast.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Wiley-liss, Div John Wiley & Sons Inc
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dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Bi-Molecular Complementation
dc.subject
Fertilization
dc.subject
Flow Cytometry
dc.subject
Mating
dc.subject
Microscopy
dc.subject
Saccharomyces Cerevisiae
dc.subject.classification
Otras Ciencias Biológicas
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dc.subject.classification
Ciencias Biológicas
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dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
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dc.title
Quantitation of yeast cell-cell fusion using multicolor flow cytometry
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2018-06-19T16:51:12Z
dc.journal.volume
87
dc.journal.number
9
dc.journal.pagination
843-854
dc.journal.pais
Estados Unidos
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dc.journal.ciudad
Nueva York
dc.description.fil
Fil: Salzman, Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Instituto Pasteur de Montevideo; Uruguay
dc.description.fil
Fil: Porro, Valentina. Instituto Pasteur de Montevideo; Uruguay
dc.description.fil
Fil: Bollati Fogolín, Mariela. Instituto Pasteur de Montevideo; Uruguay
dc.description.fil
Fil: Aguilar, Pablo Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Instituto Pasteur de Montevideo; Uruguay
dc.journal.title
Cytometry Part A
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dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1002/cyto.a.22701/abstract
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1002/cyto.a.22701
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