Fluorescent Staining and Flow Cytometry Analysis of Lymphocytes

Single-cell preparations of spleens and mesenteric lymph nodes were prepared from tissues recovered from sacrificed animals in experiment 2 [31 (link)]. Approximately 10⁶ cells were surface stained with fluorescein isothiocyanate-labeled goat anti-Syrian hamster immunoglobulin G (FITC-IgG) and phycoerythrin-labeled anti-mouse CD4 [PE-L3T4; eBiosciences (San Diego, CA)] previously shown to interact with hamster cells [31 (link),34 (link)]. Cell surface determinant data were acquired for 10⁵ cells per sample, using a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA). Lymphocytes were selected by forward scatter gating on size, and data were analyzed using the FlowJo software program (Treestar, Ashland, OR).

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Bungiro R.D., Harrison L.M., Dondji B, & Cappello M. (2022). Comparison of percutaneous vs oral infection of hamsters with the hookworm Ancylostoma ceylanicum: Parasite development, pathology and primary immune response. PLoS Neglected Tropical Diseases, 16(1), e0010098.

Publication 2022

FITC-IgG FACSCalibur flow cytometer

Animals Anti immunoglobulin Cells Fitc Fluorescein Goat Hamster Isothiocyanate Lymph nodes Lymphocytes Mesenteric Mouse Phycoerythrin Syrian hamster Tissues

Corresponding Organization : Central Washington University

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Variable analysis

dependent variables

Cell surface determinant data for 10^5 cells per sample

control variables

Sacrificed animals in experiment 2
Size of lymphocytes selected by forward scatter gating

controls

Positive control: FITC-labeled goat anti-Syrian hamster immunoglobulin G (FITC-IgG)
Positive control: phycoerythrin-labeled anti-mouse CD4 [PE-L3T4; eBiosciences (San Diego, CA)] previously shown to interact with hamster cells [31 (link),34 (link)]

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