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Experiment Overview

Repository ID: FR-FCM-ZZQ9 Experiment name: WGA 488 PBMC staining MIFlowCyt score: 97.75%
Primary researcher: Alan Stern PI/manager: Alan Stern Uploaded by: Alan Stern
Experiment dates: 2015-08-13 - 2015-08-13 Dataset uploaded: Mar 2016 Last updated: Oct 2016
Keywords: [human PBMC] [WGA] [wheat germ agglutinin ] [cell size] Manuscripts: [27768827] Cytalogo
Organizations: Icahn School of Medicine at Mount Sinai , New York, NY (United States of America)
Purpose: We have previously determined that Wheat Germ agglutinin Alexa Fluor 488 staining intensity correlates well with FSC in both HEK293 and U87 cells(FR-FCM-ZZQ7, FR-FCM-ZZQ8). A typical flow cytometry application is quantification of the proportions of granulocytes, monocytes and lymphocytes in whole blood samples based on FSC and SSC scatterplots. Since we found that WGA staining intensity correlates well with both cell size and FSC, we wanted to investigate whether it could be used as a surrogate in such whole blood analyses.
Conclusion: We gated cells according to conventional standards to determine granulocytes, monocytes and lymphocytes based on FSC vs. SSC (Fig. 1F). The cells within these conventional gates mapped very cleanly onto a scatter plot where FSC was replaced with WGA staining intensity (Fig. 1G). These data further support the notion that WGA staining intensity is a suitable metric for cell size, and suggest that it can be used interchangeably with FSC in routine flow cytometry analyses, such as quantification of granulocyte, monocyte and lymphocyte proportions in whole blood samples.
Comments: None
Funding: Funding was provided by the Icahn School of Medicine at Mount Sinai and the NIH Grants P50GM071558 (Systems Biology Center New York), R21CA196418, R01GM104184, U54HG008098 (LINCS Center), and U24AI118644 and T32 GM062754
Quality control: Unstained WGA PBMCs were utilized to determine base line PMT voltages for FITC (Ex 488, Em 530/30)


Experiment variables

Doses
· 1ug/mL WGA Specimen_003_8006wga_P1_010.fcs · Specimen_003_8007wga_P1_009.fcs · Specimen_003_8008wga_P1_008.fcs · Specimen_003_8009wga_P1_007.fcs · Specimen_003_8010wga_P1_006.fcs
· Mock Specimen_002_8006-wga_P1_005.fcs · Specimen_002_8007-wga_P1_004 - Copy.fcs · Specimen_002_8008-wga_P1_003 - Copy.fcs · Specimen_002_8009-wga_P1_002.fcs · Specimen_002_8010 -wga_P1_001.fcs

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