APPLICATION OF SIX-COLOR FLOW CYTOMETRIC ANALYSIS FOR IMMUNE PROFILE MONITORING
https://doi.org/10.15789/1563-0625-2015-1-19-26
Abstract
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+).
About the Authors
I. V. Kudryavtsev
Research Institute of Experimental Medicine, Russian Academy of Medical Sciences
197376, Russian Federation, St. Petersburg, I. Pavlov str., 12.
Far Eastern Federal University, Vladivostok, Russian Federation
Russian Federation
Russian Federation
PhD (Biology), Senior Research Associate, Department of Immunology, Institute of Experimental Medicine, North-West Branch of the Russian Academy
of Medical Sciences, St. Petersburg; Associate Professor, School of Biomedicine, Far Eastern Federal University, Vladivostok, Russian Federation
A. I. Subbotovskaya
E.N.Meshalkin Research Institute of Circulation Pathology, Russian Ministry of Health Care, Novosibirsk, Russian Federation
Russian Federation
Russian Federation
PhD (Medicine), Junior Research Associate, E.N. Meshalkin Research Institute of Circulation Pathology, Russian MInistry of Health Care, Novosibirsk, Russian Federation
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Review
For citations:
Kudryavtsev I.V., Subbotovskaya A.I. APPLICATION OF SIX-COLOR FLOW CYTOMETRIC ANALYSIS FOR IMMUNE PROFILE MONITORING. Medical Immunology (Russia). 2015;17(1):19-26. (In Russ.) https://doi.org/10.15789/1563-0625-2015-1-19-26
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