Flow Cytometry of T and B Cells and NK Cells
Posted on Jan 24, 2019
Flow cytometry can differentiate different cell populations based on cell size by forward and side scatter measurements. Specific cell types are also be identified by staining with fluorescent-tagged antibodies that recognize specific cellular markers. Readouts can be cell population analyses, proliferation, apoptosis, death/cell cycle analysis, intracellular or surface proteins, cytokine production, and binding studies.
Analysis of B and T Lymphocytes and Monocytes in Human Peripheral Blood Mononuclear Cells (PBMC)
Human PBMC, lymphocytes and monocytes, are purified and analyzed by flow cytometry. Forward scatter (FSC) measures size and side scatter (SSC) measures internal complexity (e.g. monocyte and granulocyte granules).
Figure 1 scatterplot shows at the bottom left the small lymphocytes in the bright orange region. In the upper region the larger, more granular monocytes are counted. Events at the bottom far left are a few contaminating red blood cells or platelets; these are excluded (gated out) from further analysis.
Our Attune cytometer has four lasers allowing for analysis of up to 16 different cell markers.
Figure 2 analyzes the distribution of CD3+ cells (T lymphocytes, bottom right), CD19+ cells (B lymphocytes, top left) and monocytes, CD3–CD19– (bottom left).
Figure 3 shows Natural Killer (NK) cells are in the CD56+CD16+ NK population at the top right. The bottom left contains no NK cells.
CURRENT PROJECTS INCLUDE
1. Human and animal MLR, for immunostimulation and to detect antigen-specific T cells
2. Antibody binding potency assay
3. Immune cell phenotyping
4. Multiplex ELISA to quantitate multiple cytokines in small volume samples
Tags : NK Cells, T and B Cells