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Helios cytof instrument

Manufactured by Standard BioTools
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The Helios CyTOF instrument is a mass cytometry system designed for high-dimensional single-cell analysis. It utilizes time-of-flight mass spectrometry to precisely measure the abundance of metal-tagged antibodies bound to individual cells, enabling the simultaneous detection of over 40 parameters per cell.

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Lab products found in correlation

Multimetal labeling kits, by Standard BioTools (1 mentions) Lsr 2, by BD (1 mentions) Cell id cisplatin, by Standard BioTools (1 mentions) Maxpar cell staining buffer, by Standard BioTools (1 mentions) Intercalation solution, by Standard BioTools (1 mentions) Fc receptor blocking solution, by Miltenyi Biotec (1 mentions) Intercalator ir, by Standard BioTools (1 mentions)

4 protocols using helios cytof instrument

1

Mass Cytometry Analysis of Immune Cells

Cited 3 times
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The antibodies used for mass cytometry analysis are listed in Table S2. Metal conjugated antibodies were purchased from the vendor, or conjugation was performed in house using metal-conjugation kit (Multimetal Labeling Kits, Fluidigm). All antibodies were titrated and validated for mass cytometry analysis. To control for run-to-run variation, two anchor samples were included in every mass cytometry analysis. For each analysis, 18 samples including two anchor samples (T0 and T6 mixture) and 16 samples from four subjects, four conditions each, were barcoded, combined, and surface stained as previously reported [41 (link),42 (link)]. Cells were permeabilized by Foxp3/transcription staining buffer (ebioscience, 00-5523-00), intracellularly stained, treated with DNA intercalator, and stored at 4°C until mass cytometry analysis on the following day. Data were acquired on a Helios CyTOF instrument (Fluidigm). Ten batches of mass cytometry analysis were performed over four months. Data were normalized using internal metal isotope bead standards by a previously described method [44 (link)]. Files were de-barcoded by the Matlab Debarcoder tool [44 (link)]. Acquired data were analyzed by computational analysis and manual gating.
Okamato Y., Ghosh T., Okamoto T., Schuyler R.P., Seifert J., Charry L.L., Visser A., Feser M., Fleischer C., Pedrick C., August J., Moss L., Bemis E.A., Norris J.M., Kuhn K.A., Demoruelle M.K., Deane K.D., Ghosh D., Holers V.M, & Hsieh E.W. (2020). Subjects at-risk for future development of rheumatoid arthritis demonstrate a PAD4- and TLR-dependent enhanced histone H3 citrullination and proinflammatory cytokine production in CD14hi monocytes. Journal of autoimmunity, 117, 102581.
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2

NK Cell Phenotyping and Functional Analysis

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NK cell phenotype and function was evaluated using standard protocols for flow- and mass cytometry. Samples were acquired on a BD LSRII, or LSR-Fortessa 18-color flow cytometer (BD Biosciences) or on a Helios CyTOF instrument (Fluidigm), and data were analyzed with FlowJo software V.10 (BD Biosciences) and R (R Core Team, 2019). Details are available in online supplemental materials including flow cytometry antibodies (online supplemental sTable 1) and mass cytometry antibodies (online supplemental sTable 2).
Haroun-Izquierdo A., Vincenti M., Netskar H., van Ooijen H., Zhang B., Bendzick L., Kanaya M., Momayyezi P., Li S., Wiiger M.T., Hoel H.J., Krokeide S.Z., Kremer V., Tjonnfjord G., Berggren S., Wikström K., Blomberg P., Alici E., Felices M., Önfelt B., Höglund P., Valamehr B., Ljunggren H.G., Björklund A., Hammer Q., Kveberg L., Cichocki F., Miller J.S., Malmberg K.J, & Sohlberg E. (2022). Adaptive single-KIR+NKG2C+ NK cells expanded from select superdonors show potent missing-self reactivity and efficiently control HLA-mismatched acute myeloid leukemia. Journal for Immunotherapy of Cancer, 10(11), e005577.
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3

Multiplexed CyTOF Profiling of CAR T Cells

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Purified CD22-CAR;PRODH2 and CD22-CAR;PRODH2(Stop) T cells (without cancer stimulation) were collected and washed with PBS, resuspended cell to 1e7 / mL in PBS and Cell-ID Cisplatin (Fluidigm) was added to a final concentration of 5 μM. Cells were incubated at room temperature for 5 min, then washed with Maxpar Cell Staining Buffer (Fluidigm). 1.5e6 CAR T cells per replicate were used for staining, each group has three replicates. Cells were stained with surface marker antibody cocktail first, then fixed and permeabilized. Second round staining was performed using cytoplasmic / secreted antibody cocktail. Finally, cells were incubated in intercalation solution (Fluidigm) in a final concentration of 125 nM, then incubated overnight at 4 °C. Before running on a CyTOF machine, cell concentrations were adjusted to 5–7e5/ mL with water. All data were collected on a CyTOF Helios instrument (Fluidigm). All surface and cytoplasmic / secreted antibodies were purchased from Fluidigm or Yale CyTOF core.
A list of CyTOF antibodies used in this study can be found in KRT.
Ye L., Park J.J., Peng L., Yang Q., Chow R.D., Dong M.B., Lam S.Z., Guo J., Tang E., Zhang Y., Wang G., Dai X., Du Y., Kim H.R., Cao H., Errami Y., Clark P., Bersenev A., Montgomery R.R, & Chen S. (2022). A genome-scale gain-of-function CRISPR screen in CD8 T cells identifies proline metabolism as a means to enhance CAR-T therapy. Cell metabolism, 34(4), 595-614.e14.
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4

Mass Cytometry Immunophenotyping of T Memory Cells

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The relevant metal was conjugated to each antibody according to the manufacturer’s specifications (Fluidigm Maxpar, South San Francisco, CA, USA). Antibody–metal pairs are shown in Table 1. Following isolation of CD4+CD45RO+ Tmem via magnetic bead separation, Tmem were resuspended with T-cell media containing 1:10,000 benzonase (25 U/mL), washed, blocked with Fc-receptor blocking solution (Miltenyi), and Ab cocktail was added at a volume of 50 μL to each sample for a total staining volume of 100 μL for 30 mintes at room temperature. Samples were then washed and incubated with 1:500 dilution of 25 cisplatin for a final concentration of 50 μM for 1 min at room temperature. Samples were then washed, fixed, and permeabilized (eBioscience) at 4°C overnight. The next day, Ir-intercalator (Fluidigm) was added at 1:1,000 dilution in 1 mL fixation/permeabilization butter for a final concentration of 125 nM at 4°C overnight. Cells were then washed and resuspended in MilliQ water prior to analysis using the CyTOF Helios instrument (Fluidigm). Metal-conjugated antibodies used in the study are listed in Table 1.
Wang S., Song G., Barkestani M.N., Tobiasova Z., Wang Q., Jiang Q., Lopez R., Adelekan-Kamara Y., Fan M., Pober J.S., Tellides G, & Jane-wit D. (2023). Hedgehog costimulation during ischemia-reperfusion injury potentiates cytokine and homing responses of CD4+ T cells. Frontiers in Immunology, 14, 1248027.
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