Simultaneous measurement of five‐cell surface antigens by five‐colour immunofluorescence
Multicolour immunofluorescence and flow cytometry were used for simultaneous measurement of five‐cell surface antigens on murine spleen cells. We have been able to quantitate T‐cells, T‐cell subsets, B cells, and expression of the activation marker I‐Ad from a single sample using four directly conju...
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| Veröffentlicht in: | Cytometry (New York, N.Y.) N.Y.), 1994-04, Vol.15 (4), p.371-376 |
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| container_title | Cytometry (New York, N.Y.) |
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| creator | Beavis, Andrew J. Pennline, Kenneth J. |
| description | Multicolour immunofluorescence and flow cytometry were used for simultaneous measurement of five‐cell surface antigens on murine spleen cells. We have been able to quantitate T‐cells, T‐cell subsets, B cells, and expression of the activation marker I‐Ad from a single sample using four directly conjugated monoclonal antibodies LYT2‐APC, LM‐PE, B220‐RED613, I‐Ad‐FITC and one indirect step THY1.2‐biotin/streptavidin‐Cascade Blue Three excitation wavelengths were used (488 nm, 647 nm, and U. V. 351–364 nm) for fluorescence measurements. The combination of fluorochromes used provided good resolution such that all five fluorescence signals were spectrally resolved. The percentage of cells positive for expression of a specific cell surface marker were almost identical for singlecolour samples and the five‐colour analysis, differing by only 0.3–1.5 percentage points. © 1994 Wiley‐Liss, Inc. |
| doi_str_mv | 10.1002/cyto.990150413 |
| format | Article |
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We have been able to quantitate T‐cells, T‐cell subsets, B cells, and expression of the activation marker I‐Ad from a single sample using four directly conjugated monoclonal antibodies LYT2‐APC, LM‐PE, B220‐RED613, I‐Ad‐FITC and one indirect step THY1.2‐biotin/streptavidin‐Cascade Blue Three excitation wavelengths were used (488 nm, 647 nm, and U. V. 351–364 nm) for fluorescence measurements. The combination of fluorochromes used provided good resolution such that all five fluorescence signals were spectrally resolved. The percentage of cells positive for expression of a specific cell surface marker were almost identical for singlecolour samples and the five‐colour analysis, differing by only 0.3–1.5 percentage points. © 1994 Wiley‐Liss, Inc.</description><subject>Animals</subject><subject>Antigens, Differentiation, B-Lymphocyte - analysis</subject><subject>Antigens, Differentiation, T-Lymphocyte - analysis</subject><subject>Antigens, Ly - analysis</subject><subject>Antigens, Surface - analysis</subject><subject>Flow Cytometry</subject><subject>Fluorescent Antibody Technique</subject><subject>Fluorescent Dyes</subject><subject>Histocompatibility Antigens Class II - analysis</subject><subject>Immunophenotyping - methods</subject><subject>Leukocyte Common Antigens - analysis</subject><subject>Lymphocyte Activation</subject><subject>Lymphocyte Subsets</subject><subject>Male</subject><subject>Membrane Glycoproteins - analysis</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>monoclonal antibodies</subject><subject>multicolour immunofluorescence</subject><subject>murine surface markers</subject><subject>Spleen - cytology</subject><subject>Thy-1 Antigens</subject><issn>0196-4763</issn><issn>1097-0320</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1LxDAQhoMo67p69Sb05K1rPtqkOcriFyzswfUgCCVNJ1JpmjVpld78Cf5Gf4lduqxHT8PM-87LzIPQOcFzgjG90n3r5lJikuKEsAM0JViKGDOKD9EUE8njRHB2jE5CeMMYS56wCZoISdjQTNHLY2W7ulUNuC5EFlToPFho2siZyFQf8PP1raGuo2FulIZINW31Ck2Iin6vu9p1Pqqs7Rpn6s55CBoaDafoyKg6wNmuztDT7c16cR8vV3cPi-tlrKlMWVzytCSU0qLIqNIJ45KWlOkUSCYKlhWiKIfngCsplFIcp4JjnAlTKgEsKw2bocsxd-PdewehzW0VtlePb-WCp1kmEjYY56NRexeCB5NvfGWV73OC8y3OfIsz3-McFi52yV1hodzbd_wGXY76Z1VD_09avnher_6yfwFtHIXq</recordid><startdate>19940401</startdate><enddate>19940401</enddate><creator>Beavis, Andrew J.</creator><creator>Pennline, Kenneth J.</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><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>19940401</creationdate><title>Simultaneous measurement of five‐cell surface antigens by five‐colour immunofluorescence</title><author>Beavis, Andrew J. ; Pennline, Kenneth J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c2953-d65d1222bb82ac43692d23c5e187b38b7bd504e6a97aaa605760087fda7e38df3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>Animals</topic><topic>Antigens, Differentiation, B-Lymphocyte - analysis</topic><topic>Antigens, Differentiation, T-Lymphocyte - analysis</topic><topic>Antigens, Ly - analysis</topic><topic>Antigens, Surface - analysis</topic><topic>Flow Cytometry</topic><topic>Fluorescent Antibody Technique</topic><topic>Fluorescent Dyes</topic><topic>Histocompatibility Antigens Class II - analysis</topic><topic>Immunophenotyping - methods</topic><topic>Leukocyte Common Antigens - analysis</topic><topic>Lymphocyte Activation</topic><topic>Lymphocyte Subsets</topic><topic>Male</topic><topic>Membrane Glycoproteins - analysis</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>monoclonal antibodies</topic><topic>multicolour immunofluorescence</topic><topic>murine surface markers</topic><topic>Spleen - cytology</topic><topic>Thy-1 Antigens</topic><toplevel>online_resources</toplevel><creatorcontrib>Beavis, Andrew J.</creatorcontrib><creatorcontrib>Pennline, Kenneth J.</creatorcontrib><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>Cytometry (New York, N.Y.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Beavis, Andrew J.</au><au>Pennline, Kenneth J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Simultaneous measurement of five‐cell surface antigens by five‐colour immunofluorescence</atitle><jtitle>Cytometry (New York, N.Y.)</jtitle><addtitle>Cytometry</addtitle><date>1994-04-01</date><risdate>1994</risdate><volume>15</volume><issue>4</issue><spage>371</spage><epage>376</epage><pages>371-376</pages><issn>0196-4763</issn><eissn>1097-0320</eissn><abstract>Multicolour immunofluorescence and flow cytometry were used for simultaneous measurement of five‐cell surface antigens on murine spleen cells. We have been able to quantitate T‐cells, T‐cell subsets, B cells, and expression of the activation marker I‐Ad from a single sample using four directly conjugated monoclonal antibodies LYT2‐APC, LM‐PE, B220‐RED613, I‐Ad‐FITC and one indirect step THY1.2‐biotin/streptavidin‐Cascade Blue Three excitation wavelengths were used (488 nm, 647 nm, and U. V. 351–364 nm) for fluorescence measurements. The combination of fluorochromes used provided good resolution such that all five fluorescence signals were spectrally resolved. The percentage of cells positive for expression of a specific cell surface marker were almost identical for singlecolour samples and the five‐colour analysis, differing by only 0.3–1.5 percentage points. © 1994 Wiley‐Liss, Inc.</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>7913009</pmid><doi>10.1002/cyto.990150413</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Cytometry (New York, N.Y.), 1994-04, Vol.15 (4), p.371-376 |
| issn | 0196-4763 1097-0320 |
| language | eng |
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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Animals Antigens, Differentiation, B-Lymphocyte - analysis Antigens, Differentiation, T-Lymphocyte - analysis Antigens, Ly - analysis Antigens, Surface - analysis Flow Cytometry Fluorescent Antibody Technique Fluorescent Dyes Histocompatibility Antigens Class II - analysis Immunophenotyping - methods Leukocyte Common Antigens - analysis Lymphocyte Activation Lymphocyte Subsets Male Membrane Glycoproteins - analysis Mice Mice, Inbred BALB C monoclonal antibodies multicolour immunofluorescence murine surface markers Spleen - cytology Thy-1 Antigens |
| title | Simultaneous measurement of five‐cell surface antigens by five‐colour immunofluorescence |
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