Flow Cytometry

General

An introduction to automated flow cytometry gating tools and their implementation

General Flow Cytometry protocol

Human

Regulatory T cells (Tregs)

Myeloid Derived Suppressor Cells (both granulocytic and monocytic)

  • Please note that MDSCs must be measured within 3 hours of taking blood.

Monocytes

Monocyte Maturity

Lymphocytes (T cells, B cells, NK cells)

T cells (naive, memory, TEMRA, senescent)

Neutrophils

Sorting monocyte subsets for RNAseq

 Mice

Flow cytometry protocol for staining Ly6Chigh (inflammatory)and Ly6Clow (patrolling) monocytes

Flow cytometry for bone marrow progenitors

  • This stain can be used to identify hematopoetic stem cells, common lymphoid progenitors, common myeloid progenitors, megakaryocyte-erythroid progenitors and granulocyte/monocyte progenitors

Flow cytometry protocol for myeloid populations in the lung

T cells (naive, memory, TEMRA)

Microglia and cells recruited to the brain during infection

Bacterial flow cytometry

Measurement of binding to intact bacteria

  • This protocol is used for “bacterial flow cytometry” (running bacteria through the flow cytometer rather than eukaroytic cells). We measure protein binding to the bacteria but it could easily be adapted to antibodies.