Multicolor Flow Cytometry Analysis

A sample of EDTA anticoagulated whole blood was incubated with BD Tritest CD4FITC/CD8PE/CD3PerCP reagent, another with CD3FITC/CD19PE, and the third one with CD45FITC/CD14PE (all reagents of BD Biosciences) and isotype controls, according to the manufacturer’s instructions (Díaz et al., 2015 (link)). Stained cells were analyzed with a FACSAria II flow cytometer (BD Biosciences). The percentage of positive cells and the mean fluorescence intensity (arbitrary units) for a specific marker were calculated using FACSDiva software (BD Biosciences). For each sample, 30.000 events were recorded.

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Díaz A., D’Attilio L., Penas F., Bongiovanni B., Massa E., Cevey A., Santucci N., Bottasso O., Goren N, & Bay M.L. (2023). Studies on the contribution of PPAR Gamma to tuberculosis physiopathology. Frontiers in Cellular and Infection Microbiology, 13, 1067464.

Publication 2023

BD Tritest CD4FITC/CD8PE/CD3PerCP reagent CD3FITC/CD19PE FACSAria II flow cytometer FACSDiva software

Blood Edta Fluorescence Isotype

Corresponding Organization : Consejo Nacional de Investigaciones Científicas y Técnicas

Other organizations : University of Buenos Aires, Instituto de Investigaciones Biomédicas en Retrovirus y Sida

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Variable analysis

independent variables

Incubation of whole blood samples with different fluorescent-labeled antibody reagents:
1. BD Tritest CD4FITC/CD8PE/CD3PerCP
2. CD3FITC/CD19PE
3. CD45FITC/CD14PE

dependent variables

Percentage of positive cells for a specific marker
Mean fluorescence intensity (arbitrary units) for a specific marker

control variables

EDTA anticoagulated whole blood samples
Isotype controls for the fluorescent-labeled antibody reagents
Sample size of 30,000 events recorded for each sample

controls

Positive controls: Not explicitly mentioned
Negative controls: Isotype controls for the fluorescent-labeled antibody reagents

Annotations

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