Immunology Capabilities

Mark Sangster
Director: Mark Sangster Ph.D.
Mark_Sangster@URMC.Rochester.edu

Immunofluorescent Multicolor Flow Cytometry

  • Staining and data acquisition and analysis service for immunophenotyping of cell populations.
  • Evaluation of B cell responses to immunization or infection by flow cytometry
  • Staining of cell populations for cell sorting
  • Developed and validate high-parameter staining panels for multiparameter analysis of cell populations. Established panels for the analysis of human cells include:
    • B cell activation/homing panel (CD3 CD19 CD27 CD38 CD138 CD62L CD103 Ki-67)
    • Nasal wash cell subset panel (CD3 CD4 CD8 CD11b CD11c CD45 CD56 NKp46 TCRgamma/delta MHCII)
    • CD8 T cell function panel (CD3 CD4 CD8 CD14 CD19 CD56 CD69 NKp44 NKp46 IL-2 IFN-gamma TNF-alpha)
  • Identification of antigen-specific T and B cells and function
    • In vitro stimulation and multicolor flow cytometry for cytokine detection from T cells
    • Rapid HLA Class I epitope identification and custom tetramer synthesis using HLA ligand exchange technology
    • Direct ELISpot assays on cell populations to enumerate total or antigen-specific Ab-secreting cells.
    • Measurement of antigen-specific memory B cell frequencies
  • Mass-spectrometer flow cytometry (CyTOF)
    • 35+ parameter cytometry using single-cell time-of-flight mass-spec of lanthanide labeled antibodies
  • Serological assays
    • The core has extensive experience measuring antigen-specific Ab levels in fluids using a range of techniques, such as ELISA and microneutralization assay.