Flow Cytometry Staining of Mycobacterium tuberculosis Infected Macrophage Subpopulations

Published: November 30, 2023

Abstract

Source: Mily, A., et al. Polarization of M1 and M2 Human Monocyte-Derived Cells and Analysis with Flow Cytometry upon Mycobacterium tuberculosis Infection. J. Vis. Exp. (2020)

This video showcases a flow cytometric gating strategy utilizing a multi-color panel to analyze cell viability and specific fluorescent-conjugated anti-human antibody markers. This approach makes it possible to determine the percentage of Mycobacterium tuberculosis-infected cell populations accurately.

Protocol

1. Flow cytometry staining of Mtb-infected monocyte-derived cells NOTE: The following steps must be performed in a BSL-3 facility. The flow cytometry staining could be performed in a 96-well plate instead of tubes. Detach the Mycobacterium tuberculosis, Mtb-infected cells (and uninfected controls) from the wells in the 6 well plate(s) by incubation with 1 mL of FACS buffer per well for at least 30 min at 37 °C and 5% CO2.</…

Declarações

The authors have nothing to disclose.

Materials

Formaldehyde Sigma-Aldrich F8775
Goat anti-mouse IgG Alexa Fluor 594 secondary antibody Invitrogen R37121 Secondary antibody for CD64
Goat anti-Rabbit IgG Alexa Fluor 594 secondary antibody A-11037 Secondary antibody for CD163
Fetal bovine serum (FBS) Sigma-Aldrich F7524
EDTA (0.5 M) Karolinska University hospital, Huddinge N/A
BD Comp bead plus BD 560497

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Citar este artigo
Flow Cytometry Staining of Mycobacterium tuberculosis Infected Macrophage Subpopulations. J. Vis. Exp. (Pending Publication), e21801, doi: (2023).

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