Flow Cytometric Analysis of Immune Cell Populations

Cell suspensions were blocked with FcγIII/II receptor antibody (BD), and subsequently stained with antibodies from BD bioscience (San Jose, CA, USA), i.e., PerCP-Cy5.5-conjugated anti-CD45 (30-F11), FITC-conjugated anti-CD11c (HL3), PE-conjugated anti-SiglecF (E50-2440), PE-Cy7 conjugated anti-Gr-1 (cloneRB6-8C5), APC conjugated anti-CD3e (145-2C11). Cells were acquired on a LSRII flow cytometer (BD bioscience) with data analyzed using FlowJo software (v7.6.5, Tree Star Inc., Ashland, OR, USA). Cell surface marker expression patterns were used to identify the following cell types: T cells (CD45⁺ CD3⁺), neutrophils (CD45⁺ Gr-1^high), eosinophils (CD45⁺ SiglecF⁺ CD11c^low), and alveolar macrophages (CD45⁺SiglecF⁺CD11c^high) [28 (link),29 (link),30 (link)].

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Jorquera P.A., Oakley K.E., Powell T.J., Palath N., Boyd J.G, & Tripp R.A. (2015). Layer-By-Layer Nanoparticle Vaccines Carrying the G Protein CX3C Motif Protect against RSV Infection and Disease. Vaccines, 3(4), 829-849.

Publication 2015

FcγIII/II receptor antibody LSRII flow cytometer

Alveolar macrophages Antibodies Cell types Cy5 5 Eosinophils Fitc Neutrophils T cells Tree

Corresponding Organization : University of Georgia

Other organizations : Artificial Cell Technologies (United States)

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Variable analysis

independent variables

Experimental treatment (e.g., cell stimulation, blocking of receptors)

dependent variables

Expression levels of cell surface markers (CD45, CD3, Gr-1, SiglecF, CD11c)

control variables

Cell suspensions
FcγIII/II receptor antibody (BD)
Antibodies from BD bioscience (PerCP-Cy5.5-conjugated anti-CD45, FITC-conjugated anti-CD11c, PE-conjugated anti-SiglecF, PE-Cy7 conjugated anti-Gr-1, APC conjugated anti-CD3e)
LSRII flow cytometer (BD bioscience)
FlowJo software (v7.6.5, Tree Star Inc., Ashland, OR, USA)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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