General
- Flow cytometry basics for the uninitiated. Tips and tricks for starting your own journey to becoming an expert
Antibody titrations for flow cytometry
- This describes the process of titrating antibodies to work with our Activation Induced Marker (AIM) assay, however the basic principles apply to other flow cytometry protocols.
An introduction to automated flow cytometry gating tools and their implementation
- This review describes automated gating tools for use in multi-parameter flow cytometry.
- Citation: Verschoor et al. An introduction to automated flow cytometry gating tools and their implementation. Front. Immunol. 6:380. doi: 10.3389/fimmu.2015.00380
General Flow Cytometry protocol
Human
Myeloid Derived Suppressor Cells (both granulocytic and monocytic)
- Please note that MDSCs must be measured within 3 hours of taking blood.
- This protocol is used for defining classical, intermediate and non-classical monocytes based on CD14 & CD16 expression.
- Citation: Loukov D., et al. Monocyte activation is elevated in women with knee-osteoarthritis and associated with inflammation, BMI and pain. Osteoarthritis Cartilage. 2018 Feb;26(2):255-263
Lymphocytes (T cells, B cells, NK cells)
T cells (naive, memory, TEMRA, terminally differentiated)
Sorting monocyte subsets for RNAseq
- This protocol describes a flow cytometry staining workflow to identify ancestral SARS-CoV-2 spike-specific memory B cells in human peripheral blood mononuclear cells.
T follicular helper cell (Tfh)
Murine
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
- To identify and characterize the number and activation state of microglia and infiltrating immune cells of the mouse brain. This protocol was created by combining aspects from all the references listed. Currently this protocol has only been used with a whole brain. Given the amount of recovery it may be possible to isolate sufficient cells from specific brain regions using this protocol.
Bacteria
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.