APPLICATION OF SIX-COLOR FLOW CYTOMETRIC ANALYSIS FOR IMMUNE PROFILE MONITORING
The article deals with applications of six-color flow analysis for studying the immune state parameters by means of flow cytometry. Whole blood from healthy donors was stained with combination of monoclonal antibodies, i.e., HLA-DR-FITC, CD16+56-PE, CD4-ECD, CD19-ECD, CD8-PC5.5, CD3-PC7, CD45-APC (B...
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| Format: | Article |
| Language: | Russian |
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St. Petersburg branch of the Russian Association of Allergologists and Clinical Immunologists
2015-03-01
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| Series: | Медицинская иммунология |
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| Online Access: | https://www.mimmun.ru/mimmun/article/view/803 |
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| author | I. V. Kudryavtsev A. I. Subbotovskaya |
| author_facet | I. V. Kudryavtsev A. I. Subbotovskaya |
| author_sort | I. V. Kudryavtsev |
| collection | DOAJ |
| description | The article deals with applications of six-color flow analysis for studying the immune state parameters by means of flow cytometry. Whole blood from healthy donors was stained with combination of monoclonal antibodies, i.e., HLA-DR-FITC, CD16+56-PE, CD4-ECD, CD19-ECD, CD8-PC5.5, CD3-PC7, CD45-APC (Beckman Coulter, USA) using a “no-wash” technology. To adjust the photomultiplier (PMT) voltage, we used the tubes with each of the tested monoclonal antibodies, PMT voltage was considered optimal when the negative population was located in the middle of the first decade at the logarithmic scale. The compensatory adjustment was performed in automatic mode, using Navios software (Beckman Coulter, USA). We discuss an optimal gating strategy in order to assess the populations of interest: total T cells (CD3+CD19-), T helper cells (CD3+CD4+), cytotoxic T lymphocytes (CD3+CD8+), B cells (CD3-CD19+), NK cells (CD3-CD16+CD56+), NKT cells (CD3+CD16+CD56+), activated T lymphocytes (CD3+HLA-DR+). |
| format | Article |
| id | doaj-art-5b894215711745bd80ecdec4d39b4ad3 |
| institution | Kabale University |
| issn | 1563-0625 2313-741X |
| language | Russian |
| publishDate | 2015-03-01 |
| publisher | St. Petersburg branch of the Russian Association of Allergologists and Clinical Immunologists |
| record_format | Article |
| series | Медицинская иммунология |
| spelling | doaj-art-5b894215711745bd80ecdec4d39b4ad32025-08-20T04:00:09ZrusSt. Petersburg branch of the Russian Association of Allergologists and Clinical ImmunologistsМедицинская иммунология1563-06252313-741X2015-03-01171192610.15789/1563-0625-2015-1-19-26728APPLICATION OF SIX-COLOR FLOW CYTOMETRIC ANALYSIS FOR IMMUNE PROFILE MONITORINGI. V. Kudryavtsev0A. I. Subbotovskaya1Research Institute of Experimental Medicine, Russian Academy of Medical Sciences 197376, Russian Federation, St. Petersburg, I. Pavlov str., 12. Far Eastern Federal University, Vladivostok, Russian FederationE.N.Meshalkin Research Institute of Circulation Pathology, Russian Ministry of Health Care, Novosibirsk, Russian FederationThe article deals with applications of six-color flow analysis for studying the immune state parameters by means of flow cytometry. Whole blood from healthy donors was stained with combination of monoclonal antibodies, i.e., HLA-DR-FITC, CD16+56-PE, CD4-ECD, CD19-ECD, CD8-PC5.5, CD3-PC7, CD45-APC (Beckman Coulter, USA) using a “no-wash” technology. To adjust the photomultiplier (PMT) voltage, we used the tubes with each of the tested monoclonal antibodies, PMT voltage was considered optimal when the negative population was located in the middle of the first decade at the logarithmic scale. The compensatory adjustment was performed in automatic mode, using Navios software (Beckman Coulter, USA). We discuss an optimal gating strategy in order to assess the populations of interest: total T cells (CD3+CD19-), T helper cells (CD3+CD4+), cytotoxic T lymphocytes (CD3+CD8+), B cells (CD3-CD19+), NK cells (CD3-CD16+CD56+), NKT cells (CD3+CD16+CD56+), activated T lymphocytes (CD3+HLA-DR+).https://www.mimmun.ru/mimmun/article/view/803flow cytometrymulticolor analysisimmunophenotyping |
| spellingShingle | I. V. Kudryavtsev A. I. Subbotovskaya APPLICATION OF SIX-COLOR FLOW CYTOMETRIC ANALYSIS FOR IMMUNE PROFILE MONITORING Медицинская иммунология flow cytometry multicolor analysis immunophenotyping |
| title | APPLICATION OF SIX-COLOR FLOW CYTOMETRIC ANALYSIS FOR IMMUNE PROFILE MONITORING |
| title_full | APPLICATION OF SIX-COLOR FLOW CYTOMETRIC ANALYSIS FOR IMMUNE PROFILE MONITORING |
| title_fullStr | APPLICATION OF SIX-COLOR FLOW CYTOMETRIC ANALYSIS FOR IMMUNE PROFILE MONITORING |
| title_full_unstemmed | APPLICATION OF SIX-COLOR FLOW CYTOMETRIC ANALYSIS FOR IMMUNE PROFILE MONITORING |
| title_short | APPLICATION OF SIX-COLOR FLOW CYTOMETRIC ANALYSIS FOR IMMUNE PROFILE MONITORING |
| title_sort | application of six color flow cytometric analysis for immune profile monitoring |
| topic | flow cytometry multicolor analysis immunophenotyping |
| url | https://www.mimmun.ru/mimmun/article/view/803 |
| work_keys_str_mv | AT ivkudryavtsev applicationofsixcolorflowcytometricanalysisforimmuneprofilemonitoring AT aisubbotovskaya applicationofsixcolorflowcytometricanalysisforimmuneprofilemonitoring |