Titlebook: Mass Cytometry; Methods and Protocol Helen M. McGuire,Thomas M. Ashhurst Book 2019 Springer Science+Business Media, LLC, part of Springer N

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Method for Tagging Antibodies with Metals for Mass Cytometry Experimentsodies (Abs) or other cellular probes within single cell suspensions or laser-ablated tissue sections. While many strategies exist for covalently crosslinking to proteins, the Fluidigm MaxPar. process is currently the most widely used and involves first loading a metal-chelating polymer with an eleme

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Scalable Conjugation and Characterization of Immunoglobulins with Stable Mass Isotope Reporters for mited number of metal-labeled antibodies are commercially available. Here we present optimized and scalable protocols for conjugation of lanthanide as well as bismuth ions to immunoglobulin (Ig) using a maleimide-functionalized chelating polymer and for characterization of the conjugate. The maleimi

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Surface Barcoding of Live PBMC for Multiplexed Mass Cytometryset of individual samples can be pooled and further processed and acquired as a large, single sample. For assays that require uncompromised profiling of cell-surface markers on live cells, barcoding by metal-labeled antibodies targeting cell-surface epitopes is the barcoding approach of choice. Here

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Automated Cell Processing for Mass Cytometry Experimentsprobes are coupled to elemental tags, each with a unique mass and detectable at single-cell resolution using an ICP-MS type of instrument. Given the sensitivity of the detection system, any free metal ions must be carefully removed through multiple rounds of washing in order to prevent background si

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Live Cell Barcoding for Efficient Analysis of Small Samples by Mass Cytometryh as flow cytometry. Cell stimulation and the harsh conditions required in the later stages of certain sample preparations also contribute to cell loss. Low starting cell numbers are especially susceptible to these effects, potentially limiting the ability to use mass cytometry. Here is presented a

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Multiplex MHC Class I Tetramer Combined with Intranuclear Staining by Mass Cytometrytibility complex (MHC) class I tetramers allow for a direct detection of such antigen-specific CD8. T cells. Using mass cytometry together with multiplex MHC class I tetramer staining, we are able to screen more than 1000 different antigen candidates simultaneously across tissues in health and disea

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Analysis of the Murine Bone Marrow Hematopoietic System Using Mass and Flow Cytometryan organism. Long-lived hematopoietic stem cells give rise to early progenitors with multi-lineage potential that progressively differentiate into lineage-specific progenitors. Following lineage commitment, these progenitors proliferate and expand, before eventually differentiating into their mature

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Mass Cytometric Cell Cycle Analysisevelopmental of multicellular organisms. While measurement of cell cycle state by fluorescent flow cytometry is well established, mass cytometry allows the cell cycle to be measured along with large numbers of other antigens enabling characterization of the complex interactions between the cell cycl