OMIPs Directory

OMIP-076

High-dimensional immunophenotyping of murine T-cell, B-cell, and antibody secreting cell subsets

Cytometry PART A, Volume 99, Issue 9, 888-892 (2021)
Kyle T. Mincham, Jacob D. Young, Deborah H. Strickland

Description: This 19-parameter, 18-color flow cytometry panel was designed and optimized to enable the comprehensive and simultaneous immunophenotyping of distinct T-cell, B-cell, and antibody secreting cell (ASC) subsets within murine tissues. Cellular populations identified by using this OMIP include two major subsets of B-cells (memory and activated), two ASC subsets (plasma cells and plasmablasts), and seven major subsets of CD4+ T-cells (naïve, central memory, effector memory, helper, regulatory, follicular helper, and follicular regulatory). Staining was performed on freshly isolated splenocytes from 21-day-old BALB/c mice, however, due to the omission of mouse strain-specific markers, this OMIP can be implemented across a range of murine models where in-depth immunophenotyping of the diverse repertoire of T-cell, B-cell, and ASC populations is required.

Species: Mouse

Tissue: Spleen

Cell Subset: B cell, B memory, T cell, Ctl, T helper, T memory, Tcrgd, Treg, Tfh

Machine: BD LSR Fortessa

Panel details

Marker Species Clone Fluorophore Purpose Co-expressions groups Antigen Density
CD273 Mouse TY25 Brilliant Ultraviolet 395 Memory B-cells Low
PD-1 Mouse J43 Brilliant Violet 480 T Follicular helper cells T cell, Tfh Low
FVS575V All Species FVS575V Viable cells
CD80 Mouse 16-10A1 Brilliant Violet 650 Activated B-cells B memory Low
IgM Mouse R6-60.2 Brilliant Violet 711 B-cell/ASC subsets B memory, ASC Low
CD4 Mouse RM4-5 Brilliant Violet 786 CD4+ T-cells T cell, T memory, Tfh, Treg High
CD138 Mouse 281-2 Brilliant Blue 515 Plasmablasts/Plasma cells ASC Med
TCR beta Mouse H57-597 Brilliant Blue 700 Pan T-cells T cell, Treg, Tfh, T memory Med
FOXP3 Mouse FJK-16s PE Regulatory T-cells Treg Med
CD45R Mouse RA3-6B2 PE-CF594 B-cell subsets T cell, T memory, Treg, Tfh Med
CD25 Mouse PC61 PE-Cy5 Activated T-cells T cell, Treg, Tfh Low
CD185 Mouse 2G8 PE-Cy7 T Follicular helper cells T cell, Tfh Low
MHC Class II (I-A/I-E) Mouse M5/114.15.2 (also known as M5/114) Alexa Fluor 647 B-cell subsets B memory Low
CD278 Mouse 7E.17G9 Brilliant Violet 421 T Follicular helper/Treg T cell, Treg, T memory, Tfh Low
IgD Mouse AMS 9.1 Brilliant Ultraviolet 496 Activated/memory B-cells B memory High
CD62L Mouse Mel-14 APC-R700 T-cell subsets T cell, T memory Med
CD19 Mouse 1D3 Brilliant Violet 786 B-cell subsets B memory Med
CD45 Mouse 30-F11 APC-Cy7 Pan leukocyte T cell, B memory, Tfh, Treg, T memory, ASC High
CD44 Mouse IM7 Brilliant Ultraviolet 737 T-cell subsets T cell, Treg, T memory Med