Background
Immuneprofiling by flow cytometry is essential to uncovering novel immunological mechanisms of cancer, infections, autoimmunity, and immunosenescence. Fluorescence-based flow cytometry can be challenging due to spectral overlap, autofluorescence, parameter limitation, and the large number of cells needed for single-stained controls. CyTOF® flow cytometry (CyTOF) utilizesmetal-tagged antibodies and goes beyond the limitations of fluorescence to simultaneously detect 50-plus markers per tube of sample with easy panel design and no need for single-stained or autofluorescence controls. CyTOF provides an efficient and unbiased approach to discovering novel subsets and unique functional states of immune cells, maximizing insights from precious samples.
Materials and Methods
The Maxpar® OnDemand Mouse Immune Profiling Panel Kit (Mouse Immune Profiling Panel) was designed for CyTOF to provide an efficient and comprehensive end-to-end solution for mouse immune profiling studies. The Mouse Immune Profiling Panel comprises 33 antibodies targeting key lineage and functional surface markers to characterize at least 38 lymphocyte and myeloid cell populations in mouse tissues. In support of numerous metal-tagged antibodies provided by Standard BioTools™, the Mouse Immune Profiling Panel can be customized into various high-parameter panels to meet various needs of mouse studies. Based on the Mouse Immune Profiling Panel, preset templates for automated analysis were designed by Standard BioTools to provide easy and unbiased immune profiling analysis by Maxpar Pathsetter™ software.
Results
Immunosenescenceperturbs lung cancer onset and development, yet the mechanisms remain largely unknown. To identify aging-associated immune alteration in mice, the Mouse Immune Profiling Paneland 10 Maxpar antibodies were utilized to develop a 50-parameter CyTOF panel (Figure 1A) to characterize and compare 55 pulmonary immune subsets between aged and young adult mice (Figure 1B). Maxpar Pathsetter software was used for automated high-dimensional analysis.
Conclusions
This study highlights how StandardBioTools mouse antibody products and analytical tools can be incorporated to enable comprehensive single-cell immune profiling of mouse models.
W. Xu: A. Employment (full or part-time); Significant; Standard BioTools. S. Li: A. Employment (full or part-time); Significant; Standard BioTools. A. Bouzekri: A. Employment (full or part-time); Significant; Standard BioTools. L. Tracey: A. Employment (full or part-time); Significant; Standard BioTools. C. Loh: A. Employment (full or part-time); Significant; Standard BioTools.