Experiment Overview
| Repository ID: | FR-FCM-Z4DV | Experiment name: | An Analysis of Phenotyping and Gating Strategies for B Cells in Human Peripheral Blood | MIFlowCyt score: | 59.00% |
| Primary researcher: | Kathryn Hally | PI/manager: | Kathryn Hally | Uploaded by: | Kathryn Hally |
| Experiment dates: | 2018-08-01 - 2020-12-31 | Dataset uploaded: | Aug 2021 | Last updated: | Dec 2021 |
| Keywords: | [B cells] [Phenotyping] [Flow cytometry.] [peripheral blood mononuclear cells] [gating strategy] | Manuscripts: | |||
| Organizations: | None | ||||
| Purpose: | The purpose of this experiment was to design a 9-marker, 8-colour antibody panel and gating strategy for a conventional 3-laser (violet-blue-red) flow cytometer to phenotype B cells from peripheral blood mononuclear cells (PBMCs) in humans. The panel was designed using a combination of B cell lineage markers and markers to identify specific B cell subsets. | ||||
| Conclusion: | In this experiment, we present a phenotyping panel and optimized gating strategy to identify the following B cell subsets from human PBMCs: Transitional B cells, Naïve B cells, Antibody Secreting Cells (ASC), Unswitched Memory B cells, Switched Memory B cells, IgD Only Memory B cells, IgM Only Memory B cells, Double Negative 1 B cells (DN1) and Double Negative 2 B cells (DN2). Our work demonstrates the importance of choosing a gating strategy to accurately report on the prominent B cell subsets present in human peripheral blood. | ||||
| Comments: | None | ||||
| Funding: | Not disclosed | ||||
| Quality control: | Daily instrument QC, use of application settings (on BD FACSDiva) and FMO controls for appropriate markers. | ||||
