Quantifying Antigen-Specific Cells in Murine Blood

Whole blood from euthanized mice was collected by venipuncture directly into sodium citrate (3.2% w/v) at a ratio of 9:1 v/v. Quantification of antigen-specific cell numbers in peripheral blood was measured using a whole blood assay. Blood was stained directly with fluorescent antibody cocktails as previously described^{1 (link)}. AccuCheck fluorescent beads (Invitrogen) were added to each sample before lysing the red blood cells with BD FACS lysing solution (BD Biosciences) and samples analyzed by flow cytometry as described below. Cell concentrations were determined by comparing cellular events to bead events.

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Blair T., Baird J., Bambina S., Kramer G., Gostissa M., Harvey C.J., Gough M.J, & Crittenden M.R. (2022). ICOS is upregulated on T cells following radiation and agonism combined with radiation results in enhanced tumor control. Scientific Reports, 12, 14954.

Publication 2022

AccuCheck fluorescent beads BD FACS lysing solution

Antigen Assay Blood Cellular Flow cytometry Fluorescent antibody Mice Red blood cells Sodium citrate Venipuncture

Corresponding Organization : Oregon Clinic

Other organizations : Providence Portland Medical Center

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Variable analysis

independent variables

Whole blood from euthanized mice

dependent variables

Quantification of antigen-specific cell numbers in peripheral blood

control variables

Whole blood was collected by venipuncture directly into sodium citrate (3.2% w/v) at a ratio of 9:1 v/v
AccuCheck fluorescent beads (Invitrogen) were added to each sample before lysing the red blood cells with BD FACS lysing solution (BD Biosciences)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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