Fluorimetric quantification of cell death in monolayer cultures and cell suspensions
A fluorimetric assay using ethidium bromide (E) was employed to quantify cell death in monolayer cell cultures (MA-104 cells) in situ and isolated cell suspensions (isolated colonic cells and Leishmania). Fluorescence of EB stained cells was measured with a photometer coupled to an inverted microsco...
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| Veröffentlicht in: | Journal of biochemical and biophysical methods 1991-10, Vol.23 (3), p.237-248 |
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| container_title | Journal of biochemical and biophysical methods |
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| creator | Ruiz, Marie-Christene Michelangeli, Fabián Ludert, Juan Ernesto Liprandi, Ferdinando del Castillo, Jesús R. Chemello, María Elena Benaim, Gustavo Cohen, Eleazar |
| description | A fluorimetric assay using ethidium bromide (E) was employed to quantify cell death in monolayer cell cultures (MA-104 cells) in situ and isolated cell suspensions (isolated colonic cells and
Leishmania). Fluorescence of EB stained cells was measured with a photometer coupled to an inverted microscope for cell monolayers or in a spectrofluorometer for cell suspensions. Dead cells stained with trypan blue were fluorescent with EB in all preparations studied, but the latter gave an unequivocal signal. Staining with EB and fluorescein diacetate was mutually exclusive. The relationship between the number of EB fluorescent cells and the intensity of fluorescence measured in the microphotometer was linear for a large range of cell numbers (1–14000) from different types of preparations. Applicability of the method for measuring libing and dead cells in two different time scales (minutes and hours) is shown using MA-104 cell monolayers infected with rotavirus and
Leishmania suspensions treated with amphotericin B. The method is fast, simple sensitive and reliable, enabling quantification of living and dead cells in monolayers and suspensions |
| doi_str_mv | 10.1016/0165-022X(91)90016-P |
| format | Article |
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Leishmania). Fluorescence of EB stained cells was measured with a photometer coupled to an inverted microscope for cell monolayers or in a spectrofluorometer for cell suspensions. Dead cells stained with trypan blue were fluorescent with EB in all preparations studied, but the latter gave an unequivocal signal. Staining with EB and fluorescein diacetate was mutually exclusive. The relationship between the number of EB fluorescent cells and the intensity of fluorescence measured in the microphotometer was linear for a large range of cell numbers (1–14000) from different types of preparations. Applicability of the method for measuring libing and dead cells in two different time scales (minutes and hours) is shown using MA-104 cell monolayers infected with rotavirus and
Leishmania suspensions treated with amphotericin B. The method is fast, simple sensitive and reliable, enabling quantification of living and dead cells in monolayers and suspensions</description><identifier>ISSN: 0165-022X</identifier><identifier>EISSN: 1872-857X</identifier><identifier>DOI: 10.1016/0165-022X(91)90016-P</identifier><identifier>PMID: 1779095</identifier><language>eng</language><publisher>Shannon: Elsevier B.V</publisher><subject>Animals ; Biological and medical sciences ; Cell Death - physiology ; Cell physiology ; Cell proliferation evaluation ; Cell Survival - physiology ; Cells, Cultured ; Cercopithecus aethiops ; Chemotoxicity ; Colon - cytology ; Cytotoxicity evaluation ; Ethidium ; Ethidium bromide ; Fluorometry ; Fundamental and applied biological sciences. Psychology ; In Vitro Techniques ; Isolated intestinal cell ; Leishmania ; Leishmania braziliensis - cytology ; MA-104 cell ; Microfluorometry ; Microscopy, Fluorescence ; Molecular and cellular biology ; Rats ; Reproducibility of Results ; Rotavirus ; Rotavirus Infections - pathology ; Sensitivity and Specificity</subject><ispartof>Journal of biochemical and biophysical methods, 1991-10, Vol.23 (3), p.237-248</ispartof><rights>1991</rights><rights>1992 INIST-CNRS</rights><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c337t-a153d79c4e8e1c6aa04743f177c65e6af1bb7b2b4daa266da93fffcec5c63bb23</citedby><cites>FETCH-LOGICAL-c337t-a153d79c4e8e1c6aa04743f177c65e6af1bb7b2b4daa266da93fffcec5c63bb23</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=5262188$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1779095$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ruiz, Marie-Christene</creatorcontrib><creatorcontrib>Michelangeli, Fabián</creatorcontrib><creatorcontrib>Ludert, Juan Ernesto</creatorcontrib><creatorcontrib>Liprandi, Ferdinando</creatorcontrib><creatorcontrib>del Castillo, Jesús R.</creatorcontrib><creatorcontrib>Chemello, María Elena</creatorcontrib><creatorcontrib>Benaim, Gustavo</creatorcontrib><creatorcontrib>Cohen, Eleazar</creatorcontrib><title>Fluorimetric quantification of cell death in monolayer cultures and cell suspensions</title><title>Journal of biochemical and biophysical methods</title><addtitle>J Biochem Biophys Methods</addtitle><description>A fluorimetric assay using ethidium bromide (E) was employed to quantify cell death in monolayer cell cultures (MA-104 cells) in situ and isolated cell suspensions (isolated colonic cells and
Leishmania). Fluorescence of EB stained cells was measured with a photometer coupled to an inverted microscope for cell monolayers or in a spectrofluorometer for cell suspensions. Dead cells stained with trypan blue were fluorescent with EB in all preparations studied, but the latter gave an unequivocal signal. Staining with EB and fluorescein diacetate was mutually exclusive. The relationship between the number of EB fluorescent cells and the intensity of fluorescence measured in the microphotometer was linear for a large range of cell numbers (1–14000) from different types of preparations. Applicability of the method for measuring libing and dead cells in two different time scales (minutes and hours) is shown using MA-104 cell monolayers infected with rotavirus and
Leishmania suspensions treated with amphotericin B. The method is fast, simple sensitive and reliable, enabling quantification of living and dead cells in monolayers and suspensions</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Cell Death - physiology</subject><subject>Cell physiology</subject><subject>Cell proliferation evaluation</subject><subject>Cell Survival - physiology</subject><subject>Cells, Cultured</subject><subject>Cercopithecus aethiops</subject><subject>Chemotoxicity</subject><subject>Colon - cytology</subject><subject>Cytotoxicity evaluation</subject><subject>Ethidium</subject><subject>Ethidium bromide</subject><subject>Fluorometry</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>In Vitro Techniques</subject><subject>Isolated intestinal cell</subject><subject>Leishmania</subject><subject>Leishmania braziliensis - cytology</subject><subject>MA-104 cell</subject><subject>Microfluorometry</subject><subject>Microscopy, Fluorescence</subject><subject>Molecular and cellular biology</subject><subject>Rats</subject><subject>Reproducibility of Results</subject><subject>Rotavirus</subject><subject>Rotavirus Infections - pathology</subject><subject>Sensitivity and Specificity</subject><issn>0165-022X</issn><issn>1872-857X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1991</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE1LHTEUhoNY9Gr9BxZmIUUX0yaZmWRmUyjiR0GoCwvuwpkzJ5gyN7kmM4L_vrmdi-5chBDO8x7ePIydCv5NcKG-59OUXMrH805cdDw_y_s9thKtlmXb6Md9tnpDDtlRSn8551Ur6wN2ILTueNes2MP1OIfo1jRFh8XzDH5y1iFMLvgi2AJpHIuBYHoqnC_WwYcRXikWOI_THCkV4IcFSnPakE85lz6zTxbGRCe7-5j9ub56uLwt737f_Lr8eVdiVempBNFUg-6wppYEKgBe67qyuRuqhhRY0fe6l309AEilBugqay0SNqiqvpfVMfu67N3E8DxTmszapW0Z8BTmZLRUomkVz2C9gBhDSpGs2eQ_Q3w1gputTLM1ZbamTCfMf5nmPse-7PbP_ZqG99BiL8_PdnNICKON4NGlN6yRSoq2zdiPBaPs4sVRNAkdeaTBRcLJDMF93OMfR1GTHA</recordid><startdate>199110</startdate><enddate>199110</enddate><creator>Ruiz, Marie-Christene</creator><creator>Michelangeli, Fabián</creator><creator>Ludert, Juan Ernesto</creator><creator>Liprandi, Ferdinando</creator><creator>del Castillo, Jesús R.</creator><creator>Chemello, María Elena</creator><creator>Benaim, Gustavo</creator><creator>Cohen, Eleazar</creator><general>Elsevier B.V</general><general>Elsevier Science</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>199110</creationdate><title>Fluorimetric quantification of cell death in monolayer cultures and cell suspensions</title><author>Ruiz, Marie-Christene ; Michelangeli, Fabián ; Ludert, Juan Ernesto ; Liprandi, Ferdinando ; del Castillo, Jesús R. ; Chemello, María Elena ; Benaim, Gustavo ; Cohen, Eleazar</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c337t-a153d79c4e8e1c6aa04743f177c65e6af1bb7b2b4daa266da93fffcec5c63bb23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1991</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Cell Death - physiology</topic><topic>Cell physiology</topic><topic>Cell proliferation evaluation</topic><topic>Cell Survival - physiology</topic><topic>Cells, Cultured</topic><topic>Cercopithecus aethiops</topic><topic>Chemotoxicity</topic><topic>Colon - cytology</topic><topic>Cytotoxicity evaluation</topic><topic>Ethidium</topic><topic>Ethidium bromide</topic><topic>Fluorometry</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>In Vitro Techniques</topic><topic>Isolated intestinal cell</topic><topic>Leishmania</topic><topic>Leishmania braziliensis - cytology</topic><topic>MA-104 cell</topic><topic>Microfluorometry</topic><topic>Microscopy, Fluorescence</topic><topic>Molecular and cellular biology</topic><topic>Rats</topic><topic>Reproducibility of Results</topic><topic>Rotavirus</topic><topic>Rotavirus Infections - pathology</topic><topic>Sensitivity and Specificity</topic><toplevel>online_resources</toplevel><creatorcontrib>Ruiz, Marie-Christene</creatorcontrib><creatorcontrib>Michelangeli, Fabián</creatorcontrib><creatorcontrib>Ludert, Juan Ernesto</creatorcontrib><creatorcontrib>Liprandi, Ferdinando</creatorcontrib><creatorcontrib>del Castillo, Jesús R.</creatorcontrib><creatorcontrib>Chemello, María Elena</creatorcontrib><creatorcontrib>Benaim, Gustavo</creatorcontrib><creatorcontrib>Cohen, Eleazar</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of biochemical and biophysical methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ruiz, Marie-Christene</au><au>Michelangeli, Fabián</au><au>Ludert, Juan Ernesto</au><au>Liprandi, Ferdinando</au><au>del Castillo, Jesús R.</au><au>Chemello, María Elena</au><au>Benaim, Gustavo</au><au>Cohen, Eleazar</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Fluorimetric quantification of cell death in monolayer cultures and cell suspensions</atitle><jtitle>Journal of biochemical and biophysical methods</jtitle><addtitle>J Biochem Biophys Methods</addtitle><date>1991-10</date><risdate>1991</risdate><volume>23</volume><issue>3</issue><spage>237</spage><epage>248</epage><pages>237-248</pages><issn>0165-022X</issn><eissn>1872-857X</eissn><abstract>A fluorimetric assay using ethidium bromide (E) was employed to quantify cell death in monolayer cell cultures (MA-104 cells) in situ and isolated cell suspensions (isolated colonic cells and
Leishmania). Fluorescence of EB stained cells was measured with a photometer coupled to an inverted microscope for cell monolayers or in a spectrofluorometer for cell suspensions. Dead cells stained with trypan blue were fluorescent with EB in all preparations studied, but the latter gave an unequivocal signal. Staining with EB and fluorescein diacetate was mutually exclusive. The relationship between the number of EB fluorescent cells and the intensity of fluorescence measured in the microphotometer was linear for a large range of cell numbers (1–14000) from different types of preparations. Applicability of the method for measuring libing and dead cells in two different time scales (minutes and hours) is shown using MA-104 cell monolayers infected with rotavirus and
Leishmania suspensions treated with amphotericin B. The method is fast, simple sensitive and reliable, enabling quantification of living and dead cells in monolayers and suspensions</abstract><cop>Shannon</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>1779095</pmid><doi>10.1016/0165-022X(91)90016-P</doi><tpages>12</tpages></addata></record> |
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| identifier | ISSN: 0165-022X |
| ispartof | Journal of biochemical and biophysical methods, 1991-10, Vol.23 (3), p.237-248 |
| issn | 0165-022X 1872-857X |
| language | eng |
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| subjects | Animals Biological and medical sciences Cell Death - physiology Cell physiology Cell proliferation evaluation Cell Survival - physiology Cells, Cultured Cercopithecus aethiops Chemotoxicity Colon - cytology Cytotoxicity evaluation Ethidium Ethidium bromide Fluorometry Fundamental and applied biological sciences. Psychology In Vitro Techniques Isolated intestinal cell Leishmania Leishmania braziliensis - cytology MA-104 cell Microfluorometry Microscopy, Fluorescence Molecular and cellular biology Rats Reproducibility of Results Rotavirus Rotavirus Infections - pathology Sensitivity and Specificity |
| title | Fluorimetric quantification of cell death in monolayer cultures and cell suspensions |
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