Experiment Overview
| Repository ID: | FR-FCM-ZZZ3 | Experiment name: | Identification of B cells through negative gating | MIFlowCyt score: | 89.83% |
| Primary researcher: | Karin Breuer | PI/manager: | Karin Breuer | Uploaded by: | Karin Breuer |
| Experiment dates: | 2007-05-30 - 2007-08-21 | Dataset uploaded: | Apr 2011 | Last updated: | Jan 2013 |
| Keywords: | [Innate Immune Response] [Toll-like receptors] [Activation markers] [B cells] [MIFlowCyt] | Manuscripts: | [20131398] [PMC2878765] |
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| Organizations: |
Child & Family Research Institute, Department of Pediatrics, Vancouver, BC (Canada)
University of Washington Medical Center, Department of Immunology, Seattle, Washington (USA) |
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| Purpose: | The purpose of the experiment presented here was to test whether human B cells can be identified through a negative-gating strategy. | ||||
| Conclusion: | B cells can be identified through a negative-gating strategy. Specifically, B lymphocytes are the MHC II+, CD14neg, CD11cneg, and CD123neg cells. Only on the extreme ends of a response spectrum (i.e., either none of the B cells respond to a given innate stimulation or all/most B cells respond but not monocyte, mDC or pDC) would this B cell negative-gating strategy supply acceptable data. In other words, our negative-gating strategy would potentially be difficult to interpret if both B cell and non-B cell populations responded in low frequency. | ||||
| Comments: | We set out to compare a negative-gating strategy to identify B cells with the usual positive identification of these cells using the canonical B cell surface marker, CD19. We also wanted to assess the functional response of these B cells to Toll-like receptor stimulation using the following TLR ligands: PAM3CSK4, LPS, and CpG-A ODN 2336. The blood draws, PBMC isolation, and TLR stimulations were performed between 30 May and 5 June 2007. The flow cytometry analyses were performed between 8 June and 21 August 2007. | ||||
| Funding: | Not disclosed | ||||
| Quality control: | To standardize voltage settings across samples acquired on different days, single stained controls were included. Voltages were adjusted such that fluorescence intensity was identical for each antibody, regardless of date of acquisition. | ||||
Experiment variables
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| · a2006 | a2006_O1T2pb05i_A1_A01.fcs · a2006_O1T2pb05i_A2_A02.fcs · a2006_O1T2pb05i_A3_A03.fcs · a2006_O1T2pb05i_A4_A04.fcs · a2006_O1T2pb05i_A5_A05.fcs · a2006_O1T2pb05i_A6_A06.fcs · a2006_O1T2pb05i_A7_A07.fcs · a2006_O1T2pb05i_A8_A08.fcs · a2006_O1T2pb05i_B1_B01.fcs · a2006_O1T2pb05i_B2_B02.fcs · a2006_O1T2pb05i_B3_B03.fcs · a2006_O1T2pb05i_B4_B04.fcs · a2006_O1T2pb05i_B5_B05.fcs · a2006_O1T2pb05i_B6_B06.fcs · a2006_O1T2pb05i_B7_B07.fcs · a2006_O1T2pb05i_B8_B08.fcs · a2006_O1T2pb05i_C1_C01.fcs · a2006_O1T2pb05i_C2_C02.fcs · a2006_O1T2pb05i_C3_C03.fcs · a2006_O1T2pb05i_C4_C04.fcs · a2006_O1T2pb05i_C5_C05.fcs · a2006_O1T2pb05i_C6_C06.fcs · a2006_O1T2pb05i_C7_C07.fcs · a2006_O1T2pb05i_C8_C08.fcs · a2006_O1T2pb05i_D1_D01.fcs · a2006_O1T2pb05i_D2_D02.fcs · a2006_O1T2pb05i_D3_D03.fcs · a2006_O1T2pb05i_D4_D04.fcs · a2006_O1T2pb05i_D5_D05.fcs · a2006_O1T2pb05i_D6_D06.fcs · a2006_O1T2pb05i_D7_D07.fcs · a2006_O1T2pb05i_D8_D08.fcs · a2006_O1T2pb05i_E1_E01.fcs · a2006_O1T2pb05i_E2_E02.fcs · a2006_O1T2pb05i_E3_E03.fcs · a2006_O1T2pb05i_E4_E04.fcs · a2006_O1T2pb05i_E5_E05.fcs · a2006_O1T2pb05i_E6_E06.fcs · a2006_O1T2pb05i_E7_E07.fcs · a2006_O1T2pb05i_E8_E08.fcs · a2006_O1T2pb05i_F1_F01.fcs · a2006_O1T2pb05i_F2_F02.fcs · a2006_O1T2pb05i_F3_F03.fcs · a2006_O1T2pb05i_F4_F04.fcs · a2006_O1T2pb05i_controls_G10_G10.fcs · a2006_O1T2pb05i_controls_G11_G11.fcs · a2006_O1T2pb05i_controls_G12_G12.fcs · a2006_O1T2pb05i_controls_G1_G01.fcs · a2006_O1T2pb05i_controls_G2_G02.fcs · a2006_O1T2pb05i_controls_G3_G03.fcs · a2006_O1T2pb05i_controls_G4_G04.fcs · a2006_O1T2pb05i_controls_G5_G05.fcs · a2006_O1T2pb05i_controls_G6_G06.fcs · a2006_O1T2pb05i_controls_G7_G07.fcs · a2006_O1T2pb05i_controls_G8_G08.fcs · a2006_O1T2pb05i_controls_G9_G09.fcs · a2006_O1T2pb05i_controls_H1_H01.fcs · a2006_O1T2pb05i_controls_H2_H02.fcs · a2006_O1T2pb05i_controls_H3_H03.fcs |
| · a2007 | a2007_O1T2pb05i_A1_A01.fcs · a2007_O1T2pb05i_A2_A02.fcs · a2007_O1T2pb05i_A3_A03.fcs · a2007_O1T2pb05i_A4_A04.fcs · a2007_O1T2pb05i_A5_A05.fcs · a2007_O1T2pb05i_A6_A06.fcs · a2007_O1T2pb05i_A7_A07.fcs · a2007_O1T2pb05i_A8_A08.fcs · a2007_O1T2pb05i_B1_B01.fcs · a2007_O1T2pb05i_B2_B02.fcs · a2007_O1T2pb05i_B3_B03.fcs · a2007_O1T2pb05i_B4_B04.fcs · a2007_O1T2pb05i_B5_B05.fcs · a2007_O1T2pb05i_B6_B06.fcs · a2007_O1T2pb05i_B7_B07.fcs · a2007_O1T2pb05i_B8_B08.fcs · a2007_O1T2pb05i_C1_C01.fcs · a2007_O1T2pb05i_C2_C02.fcs · a2007_O1T2pb05i_C3_C03.fcs · a2007_O1T2pb05i_C4_C04.fcs · a2007_O1T2pb05i_C5_C05.fcs · a2007_O1T2pb05i_C6_C06.fcs · a2007_O1T2pb05i_C7_C07.fcs · a2007_O1T2pb05i_C8_C08.fcs · a2007_O1T2pb05i_D1_D01.fcs · a2007_O1T2pb05i_D2_D02.fcs · a2007_O1T2pb05i_D3_D03.fcs · a2007_O1T2pb05i_D4_D04.fcs · a2007_O1T2pb05i_D5_D05.fcs · a2007_O1T2pb05i_D6_D06.fcs · a2007_O1T2pb05i_D7_D07.fcs · a2007_O1T2pb05i_D8_D08.fcs · a2007_O1T2pb05i_E1_E01.fcs · a2007_O1T2pb05i_E2_E02.fcs · a2007_O1T2pb05i_E3_E03.fcs · a2007_O1T2pb05i_E4_E04.fcs · a2007_O1T2pb05i_E5_E05.fcs · a2007_O1T2pb05i_E6_E06.fcs · a2007_O1T2pb05i_E7_E07.fcs · a2007_O1T2pb05i_E8_E08.fcs · a2007_O1T2pb05i_F1_F01.fcs · a2007_O1T2pb05i_F2_F02.fcs · a2007_O1T2pb05i_controls_G10_G10.fcs · a2007_O1T2pb05i_controls_G11_G11.fcs · a2007_O1T2pb05i_controls_G12_G12.fcs · a2007_O1T2pb05i_controls_G1_G01.fcs · a2007_O1T2pb05i_controls_G2_G02.fcs · a2007_O1T2pb05i_controls_G3_G03.fcs · a2007_O1T2pb05i_controls_G4_G04.fcs · a2007_O1T2pb05i_controls_G5_G05.fcs · a2007_O1T2pb05i_controls_G6_G06.fcs · a2007_O1T2pb05i_controls_G7_G07.fcs · a2007_O1T2pb05i_controls_G8_G08.fcs · a2007_O1T2pb05i_controls_G9_G09.fcs · a2007_O1T2pb05i_controls_H1_H01.fcs · a2007_O1T2pb05i_controls_H2_H02.fcs · a2007_O1T2pb05i_controls_H3_H03.fcs |
| · a2008 | a2008_O1T2pb05i_A1_A01.fcs · a2008_O1T2pb05i_A2_A02.fcs · a2008_O1T2pb05i_A3_A03.fcs · a2008_O1T2pb05i_A4_A04.fcs · a2008_O1T2pb05i_A5_A05.fcs · a2008_O1T2pb05i_A6_A06.fcs · a2008_O1T2pb05i_A7_A07.fcs · a2008_O1T2pb05i_A8_A08.fcs · a2008_O1T2pb05i_B1_B01.fcs · a2008_O1T2pb05i_B2_B02.fcs · a2008_O1T2pb05i_B3_B03.fcs · a2008_O1T2pb05i_B4_B04.fcs · a2008_O1T2pb05i_B5_B05.fcs · a2008_O1T2pb05i_B6_B06.fcs · a2008_O1T2pb05i_B7_B07.fcs · a2008_O1T2pb05i_B8_B08.fcs · a2008_O1T2pb05i_C1_C01.fcs · a2008_O1T2pb05i_C2_C02.fcs · a2008_O1T2pb05i_C3_C03.fcs · a2008_O1T2pb05i_C4_C04.fcs · a2008_O1T2pb05i_C5_C05.fcs · a2008_O1T2pb05i_C6_C06.fcs · a2008_O1T2pb05i_C7_C07.fcs · a2008_O1T2pb05i_C8_C08.fcs · a2008_O1T2pb05i_D1_D01.fcs · a2008_O1T2pb05i_D2_D02.fcs · a2008_O1T2pb05i_D3_D03.fcs · a2008_O1T2pb05i_D4_D04.fcs · a2008_O1T2pb05i_D5_D05.fcs · a2008_O1T2pb05i_D6_D06.fcs · a2008_O1T2pb05i_D7_D07.fcs · a2008_O1T2pb05i_D8_D08.fcs · a2008_O1T2pb05i_E1_E01.fcs · a2008_O1T2pb05i_E2_E02.fcs · a2008_O1T2pb05i_E3_E03.fcs · a2008_O1T2pb05i_E4_E04.fcs · a2008_O1T2pb05i_E5_E05.fcs · a2008_O1T2pb05i_E6_E06.fcs · a2008_O1T2pb05i_E7_E07.fcs · a2008_O1T2pb05i_E8_E08.fcs · a2008_O1T2pb05i_F1_F01.fcs · a2008_O1T2pb05i_F2_F02.fcs · a2008_O1T2pb05i_controls_G10_G10.fcs · a2008_O1T2pb05i_controls_G11_G11.fcs · a2008_O1T2pb05i_controls_G12_G12.fcs · a2008_O1T2pb05i_controls_G1_G01.fcs · a2008_O1T2pb05i_controls_G2_G02.fcs · a2008_O1T2pb05i_controls_G3_G03.fcs · a2008_O1T2pb05i_controls_G4_G04.fcs · a2008_O1T2pb05i_controls_G5_G05.fcs · a2008_O1T2pb05i_controls_G6_G06.fcs · a2008_O1T2pb05i_controls_G7_G07.fcs · a2008_O1T2pb05i_controls_G8_G08.fcs · a2008_O1T2pb05i_controls_G9_G09.fcs · a2008_O1T2pb05i_controls_H1_H01.fcs · a2008_O1T2pb05i_controls_H2_H02.fcs · a2008_O1T2pb05i_controls_H3_H03.fcs |
| · a2009 | a2009_O1T2pb05i_A1_A01.fcs · a2009_O1T2pb05i_A2_A02.fcs · a2009_O1T2pb05i_A3_A03.fcs · a2009_O1T2pb05i_A4_A04.fcs · a2009_O1T2pb05i_A5_A05.fcs · a2009_O1T2pb05i_A6_A06.fcs · a2009_O1T2pb05i_A7_A07.fcs · a2009_O1T2pb05i_A8_A08.fcs · a2009_O1T2pb05i_B1_B01.fcs · a2009_O1T2pb05i_B2_B02.fcs · a2009_O1T2pb05i_B3_B03.fcs · a2009_O1T2pb05i_B4_B04.fcs · a2009_O1T2pb05i_B5_B05.fcs · a2009_O1T2pb05i_B6_B06.fcs · a2009_O1T2pb05i_B7_B07.fcs · a2009_O1T2pb05i_B8_B08.fcs · a2009_O1T2pb05i_C1_C01.fcs · a2009_O1T2pb05i_C2_C02.fcs · a2009_O1T2pb05i_C3_C03.fcs · a2009_O1T2pb05i_C4_C04.fcs · a2009_O1T2pb05i_C5_C05.fcs · a2009_O1T2pb05i_C6_C06.fcs · a2009_O1T2pb05i_C7_C07.fcs · a2009_O1T2pb05i_C8_C08.fcs · a2009_O1T2pb05i_D1_D01.fcs · a2009_O1T2pb05i_D2_D02.fcs · a2009_O1T2pb05i_D3_D03.fcs · a2009_O1T2pb05i_D4_D04.fcs · a2009_O1T2pb05i_D5_D05.fcs · a2009_O1T2pb05i_D6_D06.fcs · a2009_O1T2pb05i_D7_D07.fcs · a2009_O1T2pb05i_D8_D08.fcs · a2009_O1T2pb05i_E1_E01.fcs · a2009_O1T2pb05i_E2_E02.fcs · a2009_O1T2pb05i_E3_E03.fcs · a2009_O1T2pb05i_E4_E04.fcs · a2009_O1T2pb05i_E5_E05.fcs · a2009_O1T2pb05i_E6_E06.fcs · a2009_O1T2pb05i_E7_E07.fcs · a2009_O1T2pb05i_E8_E08.fcs · a2009_O1T2pb05i_F1_F01.fcs · a2009_O1T2pb05i_F2_F02.fcs · a2009_O1T2pb05i_controls_G10_G10.fcs · a2009_O1T2pb05i_controls_G11_G11.fcs · a2009_O1T2pb05i_controls_G12_G12.fcs · a2009_O1T2pb05i_controls_G1_G01.fcs · a2009_O1T2pb05i_controls_G2_G02.fcs · a2009_O1T2pb05i_controls_G3_G03.fcs · a2009_O1T2pb05i_controls_G4_G04.fcs · a2009_O1T2pb05i_controls_G5_G05.fcs · a2009_O1T2pb05i_controls_G6_G06.fcs · a2009_O1T2pb05i_controls_G7_G07.fcs · a2009_O1T2pb05i_controls_G8_G08.fcs · a2009_O1T2pb05i_controls_G9_G09.fcs · a2009_O1T2pb05i_controls_H1_H01.fcs · a2009_O1T2pb05i_controls_H2_H02.fcs · a2009_O1T2pb05i_controls_H3_H03.fcs |
| · a2010 | a2010_O1T2pb05i_A1_A01.fcs · a2010_O1T2pb05i_A2_A02.fcs · a2010_O1T2pb05i_A3_A03.fcs · a2010_O1T2pb05i_A4_A04.fcs · a2010_O1T2pb05i_A8_A08.fcs · a2010_O1T2pb05i_B1_B01.fcs · a2010_O1T2pb05i_B2_B02.fcs · a2010_O1T2pb05i_B3_B03.fcs · a2010_O1T2pb05i_B4_B04.fcs · a2010_O1T2pb05i_B5_B05.fcs · a2010_O1T2pb05i_B6_B06.fcs · a2010_O1T2pb05i_B7_B07.fcs · a2010_O1T2pb05i_B8_B08.fcs · a2010_O1T2pb05i_C1_C01.fcs · a2010_O1T2pb05i_C2_C02.fcs · a2010_O1T2pb05i_C3_C03.fcs · a2010_O1T2pb05i_C4_C04.fcs · a2010_O1T2pb05i_C5_C05.fcs · a2010_O1T2pb05i_C6_C06.fcs · a2010_O1T2pb05i_C7_C07.fcs · a2010_O1T2pb05i_C8_C08.fcs · a2010_O1T2pb05i_D1_D01.fcs · a2010_O1T2pb05i_D2_D02.fcs · a2010_O1T2pb05i_D3_D03.fcs · a2010_O1T2pb05i_D4_D04.fcs · a2010_O1T2pb05i_D5_D05.fcs · a2010_O1T2pb05i_D6_D06.fcs · a2010_O1T2pb05i_D7_D07.fcs · a2010_O1T2pb05i_D8_D08.fcs · a2010_O1T2pb05i_E1_E01.fcs · a2010_O1T2pb05i_E2_E02.fcs · a2010_O1T2pb05i_E3_E03.fcs · a2010_O1T2pb05i_E4_E04.fcs · a2010_O1T2pb05i_E5_E05.fcs · a2010_O1T2pb05i_E6_E06.fcs · a2010_O1T2pb05i_E7_E07.fcs · a2010_O1T2pb05i_E8_E08.fcs · a2010_O1T2pb05i_F1_F01.fcs · a2010_O1T2pb05i_F2_F02.fcs · a2010_O1T2pb05i_controls_G10_G10.fcs · a2010_O1T2pb05i_controls_G11_G11.fcs · a2010_O1T2pb05i_controls_G12_G12.fcs · a2010_O1T2pb05i_controls_G1_G01.fcs · a2010_O1T2pb05i_controls_G2_G02.fcs · a2010_O1T2pb05i_controls_G3_G03.fcs · a2010_O1T2pb05i_controls_G4_G04.fcs · a2010_O1T2pb05i_controls_G5_G05.fcs · a2010_O1T2pb05i_controls_G6_G06.fcs · a2010_O1T2pb05i_controls_G7_G07.fcs · a2010_O1T2pb05i_controls_G8_G08.fcs · a2010_O1T2pb05i_controls_G9_G09.fcs · a2010_O1T2pb05i_controls_H1_H01.fcs · a2010_O1T2pb05i_controls_H2_H02.fcs · a2010_O1T2pb05i_controls_H3_H03.fcs |
