Perceiving Mitosis in Eukaryotic Cells
A sensitive method has been developed for visualizing eukaryotic cells in mitosis (M) phase. It employs Zenker's fixative, which makes the plasma membrane but not the nuclear envelope permeable to immunoglobulins. Zenker's-fixed cells are exposed to an antibody which recognizes a major con...
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| Veröffentlicht in: | In Vitro Cellular & Developmental Biology 1988-02, Vol.24 (2), p.100-107 |
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| container_title | In Vitro Cellular & Developmental Biology |
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| creator | Kim, Hae-Young Dennis Byrne Paul Hwang Sandra Collins Thompson Kitos, Paul A. |
| description | A sensitive method has been developed for visualizing eukaryotic cells in mitosis (M) phase. It employs Zenker's fixative, which makes the plasma membrane but not the nuclear envelope permeable to immunoglobulins. Zenker's-fixed cells are exposed to an antibody which recognizes a major constituent of chromatin. In this case the antibody is a monoclonal (MC 21) which recognizes histone H2b. Because cells in M phase do not have an intact nuclear envelope, the antibody has access to and interacts with their chromatin. The presence of a nuclear envelope in Zenker's-fixed interphase cells precludes access of the antibody to the nuclear chromatin. Consequently, this indirect immunofluorescence procedure selectively labels M-phase cells. At high enough magnification some details of the chromatin figures are revealed. MC 21 recognizes the chromatin of cells of many different species. With appropriate fixation it can be used effectively on cells in culture. With some procedural modifications it can also be used with more complex tissue systems. Detailed mitotic patterns for chick embryos up to Day 3 of development have been obtained by this method. |
| doi_str_mv | 10.1007/bf02623886 |
| format | Article |
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It employs Zenker's fixative, which makes the plasma membrane but not the nuclear envelope permeable to immunoglobulins. Zenker's-fixed cells are exposed to an antibody which recognizes a major constituent of chromatin. In this case the antibody is a monoclonal (MC 21) which recognizes histone H2b. Because cells in M phase do not have an intact nuclear envelope, the antibody has access to and interacts with their chromatin. The presence of a nuclear envelope in Zenker's-fixed interphase cells precludes access of the antibody to the nuclear chromatin. Consequently, this indirect immunofluorescence procedure selectively labels M-phase cells. At high enough magnification some details of the chromatin figures are revealed. MC 21 recognizes the chromatin of cells of many different species. With appropriate fixation it can be used effectively on cells in culture. With some procedural modifications it can also be used with more complex tissue systems. Detailed mitotic patterns for chick embryos up to Day 3 of development have been obtained by this method.</description><identifier>ISSN: 0883-8364</identifier><identifier>EISSN: 2327-431X</identifier><identifier>EISSN: 1475-2689</identifier><identifier>DOI: 10.1007/bf02623886</identifier><identifier>PMID: 3277936</identifier><identifier>CODEN: ICDBEO</identifier><language>eng</language><publisher>Largo, MD: Tissue Culture Association, Inc</publisher><subject>Animal cells ; Animals ; Antibodies ; Antibodies, Monoclonal - immunology ; Antibodies, Monoclonal - metabolism ; Biological and medical sciences ; Cell Line ; Cell lines ; Cell Membrane - metabolism ; Cell nucleus ; Cells, Cultured ; Chick Embryo ; Chromates ; Chromatin ; Chromatin - immunology ; Chromatin. Chromosome ; Cultured cells ; Drug Combinations ; Embryos ; Eukaryotic Cells - cytology ; Fibroblasts ; Fixatives ; Fluorescent Antibody Technique ; Fundamental and applied biological sciences. Psychology ; Hemostatics ; Histones ; Histones - analysis ; Histones - immunology ; Humans ; Hybridomas ; Immunoassay ; Immunoglobulins - immunology ; Immunoglobulins - metabolism ; L cells ; Mercuric Chloride ; Mitosis ; Molecular and cellular biology ; Molecular genetics ; Nuclear membrane ; Species Specificity ; Sulfates ; Tumor Cells, Cultured</subject><ispartof>In Vitro Cellular & Developmental Biology, 1988-02, Vol.24 (2), p.100-107</ispartof><rights>Copyright 1988 Tissue Culture Association, Inc.</rights><rights>1989 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c332t-691783bef1307007e9784dbbd3545df45c0226d2ae62eeec0597a201057d1b403</citedby><cites>FETCH-LOGICAL-c332t-691783bef1307007e9784dbbd3545df45c0226d2ae62eeec0597a201057d1b403</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/4296183$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/4296183$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>314,780,784,803,27923,27924,58016,58249</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=6980911$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/3277936$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kim, Hae-Young</creatorcontrib><creatorcontrib>Dennis Byrne</creatorcontrib><creatorcontrib>Paul Hwang</creatorcontrib><creatorcontrib>Sandra Collins Thompson</creatorcontrib><creatorcontrib>Kitos, Paul A.</creatorcontrib><title>Perceiving Mitosis in Eukaryotic Cells</title><title>In Vitro Cellular & Developmental Biology</title><addtitle>In Vitro Cell Dev Biol</addtitle><description>A sensitive method has been developed for visualizing eukaryotic cells in mitosis (M) phase. It employs Zenker's fixative, which makes the plasma membrane but not the nuclear envelope permeable to immunoglobulins. Zenker's-fixed cells are exposed to an antibody which recognizes a major constituent of chromatin. In this case the antibody is a monoclonal (MC 21) which recognizes histone H2b. Because cells in M phase do not have an intact nuclear envelope, the antibody has access to and interacts with their chromatin. The presence of a nuclear envelope in Zenker's-fixed interphase cells precludes access of the antibody to the nuclear chromatin. Consequently, this indirect immunofluorescence procedure selectively labels M-phase cells. At high enough magnification some details of the chromatin figures are revealed. MC 21 recognizes the chromatin of cells of many different species. With appropriate fixation it can be used effectively on cells in culture. With some procedural modifications it can also be used with more complex tissue systems. Detailed mitotic patterns for chick embryos up to Day 3 of development have been obtained by this method.</description><subject>Animal cells</subject><subject>Animals</subject><subject>Antibodies</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>Antibodies, Monoclonal - metabolism</subject><subject>Biological and medical sciences</subject><subject>Cell Line</subject><subject>Cell lines</subject><subject>Cell Membrane - metabolism</subject><subject>Cell nucleus</subject><subject>Cells, Cultured</subject><subject>Chick Embryo</subject><subject>Chromates</subject><subject>Chromatin</subject><subject>Chromatin - immunology</subject><subject>Chromatin. Chromosome</subject><subject>Cultured cells</subject><subject>Drug Combinations</subject><subject>Embryos</subject><subject>Eukaryotic Cells - cytology</subject><subject>Fibroblasts</subject><subject>Fixatives</subject><subject>Fluorescent Antibody Technique</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hemostatics</subject><subject>Histones</subject><subject>Histones - analysis</subject><subject>Histones - immunology</subject><subject>Humans</subject><subject>Hybridomas</subject><subject>Immunoassay</subject><subject>Immunoglobulins - immunology</subject><subject>Immunoglobulins - metabolism</subject><subject>L cells</subject><subject>Mercuric Chloride</subject><subject>Mitosis</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Nuclear membrane</subject><subject>Species Specificity</subject><subject>Sulfates</subject><subject>Tumor Cells, Cultured</subject><issn>0883-8364</issn><issn>2327-431X</issn><issn>1475-2689</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1988</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo90MFLwzAUBvAgypzTi2eFHmQHofqSlybpUcemwkQPCt5KmqaS2bUzaQX_eztWd3qH78fj4yPknMINBZC3eQlMMFRKHJAxQyZjjvTjkIxBKYwVCn5MTkJYASAIxkZk1BuZohiT6av1xrofV39Gz65tgguRq6N596X9b9M6E81sVYVTclTqKtiz4U7I-2L-NnuMly8PT7O7ZWwQWRuLlEqFuS0pguyb2VQqXuR5gQlPipInBhgTBdNWMGutgSSVmgGFRBY054ATMt393fjmu7OhzdYumL6Brm3ThUwqyjiILbzeQeObELwts413675zRiHbjpLdL_5H6fHl8LXL17bY02GFPr8ach2Mrkqva-PCnolUQUppzy52bBXaxu9jzlJBFeIf9Q9vwA</recordid><startdate>19880201</startdate><enddate>19880201</enddate><creator>Kim, Hae-Young</creator><creator>Dennis Byrne</creator><creator>Paul Hwang</creator><creator>Sandra Collins Thompson</creator><creator>Kitos, Paul A.</creator><general>Tissue Culture Association, Inc</general><general>Society for In Vitro Biology</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>19880201</creationdate><title>Perceiving Mitosis in Eukaryotic Cells</title><author>Kim, Hae-Young ; Dennis Byrne ; Paul Hwang ; Sandra Collins Thompson ; Kitos, Paul A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c332t-691783bef1307007e9784dbbd3545df45c0226d2ae62eeec0597a201057d1b403</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1988</creationdate><topic>Animal cells</topic><topic>Animals</topic><topic>Antibodies</topic><topic>Antibodies, Monoclonal - immunology</topic><topic>Antibodies, Monoclonal - metabolism</topic><topic>Biological and medical sciences</topic><topic>Cell Line</topic><topic>Cell lines</topic><topic>Cell Membrane - metabolism</topic><topic>Cell nucleus</topic><topic>Cells, Cultured</topic><topic>Chick Embryo</topic><topic>Chromates</topic><topic>Chromatin</topic><topic>Chromatin - immunology</topic><topic>Chromatin. Chromosome</topic><topic>Cultured cells</topic><topic>Drug Combinations</topic><topic>Embryos</topic><topic>Eukaryotic Cells - cytology</topic><topic>Fibroblasts</topic><topic>Fixatives</topic><topic>Fluorescent Antibody Technique</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Hemostatics</topic><topic>Histones</topic><topic>Histones - analysis</topic><topic>Histones - immunology</topic><topic>Humans</topic><topic>Hybridomas</topic><topic>Immunoassay</topic><topic>Immunoglobulins - immunology</topic><topic>Immunoglobulins - metabolism</topic><topic>L cells</topic><topic>Mercuric Chloride</topic><topic>Mitosis</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Nuclear membrane</topic><topic>Species Specificity</topic><topic>Sulfates</topic><topic>Tumor Cells, Cultured</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kim, Hae-Young</creatorcontrib><creatorcontrib>Dennis Byrne</creatorcontrib><creatorcontrib>Paul Hwang</creatorcontrib><creatorcontrib>Sandra Collins Thompson</creatorcontrib><creatorcontrib>Kitos, Paul A.</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>In Vitro Cellular & Developmental Biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kim, Hae-Young</au><au>Dennis Byrne</au><au>Paul Hwang</au><au>Sandra Collins Thompson</au><au>Kitos, Paul A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Perceiving Mitosis in Eukaryotic Cells</atitle><jtitle>In Vitro Cellular & Developmental Biology</jtitle><addtitle>In Vitro Cell Dev Biol</addtitle><date>1988-02-01</date><risdate>1988</risdate><volume>24</volume><issue>2</issue><spage>100</spage><epage>107</epage><pages>100-107</pages><issn>0883-8364</issn><eissn>2327-431X</eissn><eissn>1475-2689</eissn><coden>ICDBEO</coden><abstract>A sensitive method has been developed for visualizing eukaryotic cells in mitosis (M) phase. It employs Zenker's fixative, which makes the plasma membrane but not the nuclear envelope permeable to immunoglobulins. Zenker's-fixed cells are exposed to an antibody which recognizes a major constituent of chromatin. In this case the antibody is a monoclonal (MC 21) which recognizes histone H2b. Because cells in M phase do not have an intact nuclear envelope, the antibody has access to and interacts with their chromatin. The presence of a nuclear envelope in Zenker's-fixed interphase cells precludes access of the antibody to the nuclear chromatin. Consequently, this indirect immunofluorescence procedure selectively labels M-phase cells. At high enough magnification some details of the chromatin figures are revealed. MC 21 recognizes the chromatin of cells of many different species. With appropriate fixation it can be used effectively on cells in culture. With some procedural modifications it can also be used with more complex tissue systems. Detailed mitotic patterns for chick embryos up to Day 3 of development have been obtained by this method.</abstract><cop>Largo, MD</cop><pub>Tissue Culture Association, Inc</pub><pmid>3277936</pmid><doi>10.1007/bf02623886</doi><tpages>8</tpages></addata></record> |
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| subjects | Animal cells Animals Antibodies Antibodies, Monoclonal - immunology Antibodies, Monoclonal - metabolism Biological and medical sciences Cell Line Cell lines Cell Membrane - metabolism Cell nucleus Cells, Cultured Chick Embryo Chromates Chromatin Chromatin - immunology Chromatin. Chromosome Cultured cells Drug Combinations Embryos Eukaryotic Cells - cytology Fibroblasts Fixatives Fluorescent Antibody Technique Fundamental and applied biological sciences. Psychology Hemostatics Histones Histones - analysis Histones - immunology Humans Hybridomas Immunoassay Immunoglobulins - immunology Immunoglobulins - metabolism L cells Mercuric Chloride Mitosis Molecular and cellular biology Molecular genetics Nuclear membrane Species Specificity Sulfates Tumor Cells, Cultured |
| title | Perceiving Mitosis in Eukaryotic Cells |
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