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Cell id intercalator ir solution

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The Cell-ID Intercalator-Ir solution is a laboratory reagent used in cellular analysis applications. It is a metal-based DNA intercalator that binds to nucleic acids, allowing for the identification and discrimination of individual cells within a sample.

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Lab products found in correlation

Cytof software, by Standard BioTools (2 mentions) Cell id cisplatin, by Standard BioTools (2 mentions) Fc receptor binding inhibitor, by Thermo Fisher Scientific (2 mentions) Heavy metal conjugated antibodies, by Standard BioTools (2 mentions) Metal conjugated antibodies, by Standard BioTools (2 mentions) Methanol free formaldehyde, by Thermo Fisher Scientific (2 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (2 mentions) Maxpar fix and perm buffer, by Standard BioTools (1 mentions) Eq calibration beads, by Standard BioTools (1 mentions) Helios mass cytometry, by Standard BioTools (1 mentions) Cytobank platform, by Beckman Coulter (1 mentions) Formaldehyde solution, by Thermo Fisher Scientific (1 mentions) Cisplatin, by Standard BioTools (1 mentions) Helios mass cytometer, by Standard BioTools (1 mentions) Cytobank, by Beckman Coulter (1 mentions) Eq four element calibration beads, by Standard BioTools (1 mentions) Facsaria 3, by BD (1 mentions)

4 protocols using cell id intercalator ir solution

1

CyTOF Profiling of Surface and Intracellular Markers

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Frozen cells were thawed at 37°C and washed with PBS twice. Single cell suspensions were stained with Cell-ID Cisplatin (Fluidigm), blocked with Fc Receptor Binding Inhibitor (Thermo Fisher), and stained with heavy metal-conjugated antibodies (Fluidigm) for surface staining. Cells were fixed and permeabilized with FoxP3 Transcription Factor Staining Buffer Set (Thermo Fisher) followed by intracellular staining with metal-conjugated antibodies (Fluidigm). Cells were fixed in methanol-free formaldehyde (Thermo Fisher) and kept in Cell-ID Intercalator-Ir solution (Fluidigm). Stained cells were shipped on ice to the CyTOF Facility at the University of Pennsylvania and acquired on Helios (Fluidigm). CyTOF data were normalized in the CyTOF software (Fluidigm) and analyzed using Cytobank (Cytobank Inc).
Choi H.S., Won T., Hou X., Chen G., Bracamonte-Baran W., Talor M.V., Jurčová I., Szárszoi O., Čurnova L., Stříž I., Hooper J.E., Melenovský V, & Čiháková D. (2020). Innate Lymphoid Cells Play a Pathogenic Role in Pericarditis. Cell reports, 30(9), 2989-3003.e6.
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2

CyTOF Profiling of Surface and Intracellular Markers

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Frozen cells were thawed at 37°C and washed with PBS twice. Single cell suspensions were stained with Cell-ID Cisplatin (Fluidigm), blocked with Fc Receptor Binding Inhibitor (Thermo Fisher), and stained with heavy metal-conjugated antibodies (Fluidigm) for surface staining. Cells were fixed and permeabilized with FoxP3 Transcription Factor Staining Buffer Set (Thermo Fisher) followed by intracellular staining with metal-conjugated antibodies (Fluidigm). Cells were fixed in methanol-free formaldehyde (Thermo Fisher) and kept in Cell-ID Intercalator-Ir solution (Fluidigm). Stained cells were shipped on ice to the CyTOF Facility at the University of Pennsylvania and acquired on Helios (Fluidigm). CyTOF data were normalized in the CyTOF software (Fluidigm) and analyzed using Cytobank (Cytobank Inc).
Choi H.S., Won T., Hou X., Chen G., Bracamonte-Baran W., Talor M.V., Jurčová I., Szárszoi O., Čurnova L., Stříž I., Hooper J.E., Melenovský V, & Čiháková D. (2020). Innate Lymphoid Cells Play a Pathogenic Role in Pericarditis. Cell reports, 30(9), 2989-3003.e6.
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3

CyTOF Immunoprofiling of Fresh Blood

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Fresh blood was processed and stained using the Maxpar Direct Immunoprofiling Assay (MDIPA) (Fluidigm or The Longwood Medical Area CyTOF core, Boston, MA). Following processing and staining, samples were stored at −80° C. The frozen stained blood samples were placed at room temperature (RT) for 30–50 min until fully thawed and then incubated with 1× Thaw-Lyse buffer (Smart Tube Inc.) at RT for 10 min. The lysis steps were repeated up to 3 times until the pellet turned white evidencing complete lysis of red blood cells. The cells were then incubated with Maxpar Fix and Perm Buffer (Fluidigm) containing 125 nM Cell-ID Intercalator-Ir solution (Fluidigm) at 4° C overnight or up to 48 h before sample acquisition. Groups of samples (4–8/day) were assessed by Helios mass cytometry (Fluidigm) in 4 independent experiments using a flow rate of 45 μI/min in the presence of EQ Calibration beads (Fluidigm) for normalization. An average of 400,000 ± 50,000 cells (mean ± SEM) from each sample were acquired and analyzed by Helios. Gating was performed on the Cytobank platform (Cytobank, Inc. Santa Clara, CA) and FlowJo 10.5.3 (BD Biosciences).
Mi Q.S., Dimitrion P., Hamzavi I., Yin C., Loveless I., Toor J., Subedi K., Huggins R., Khalasawi N., Adrianto I., Veenstra J., Vellaichamy G., Hans A., Daveluy S., Athar M., Liao W., Lim H., Ozog D, & Zhou L. (2023). Dysregulated CD38 expression in blood and skin immune cells of patients with hidradenitis suppurativa. Research Square.
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4

Multiparameter Flow Cytometry of PKT Tumors

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Single-cell suspensions from PKT tumors were flow sorted using FACS Aria III (BD Life Sciences) to remove non-cellular contaminants. Samples were resuspended and viability staining performed using 2.5 μM cisplatin (Fluidigm) working solution following by FcR blocking and surface staining for 30 min at room temperature using tagged antibodies as shown in Table S2. Fixation was then performed using 1.6% formaldehyde solution (ThermoFisher). Cells were suspended in 125 nM Cell-ID Intercalator-Ir solution (Fluidigm) overnight at 4°C. The following day, cells were washed and resuspended in 0.1X EQ Four Element Calibration Beads (Fluidigm) before acquisition. Data were acquired by Helios Mass Cytometer (Fluidigm) and analyzed using FlowJo v10 and Cytobank (Beckman Coulter). To generate viSNE plots, individual files from each treatment group were concatenated into a single file and then a subset of equal events were selected at random through FlowJo DownSample plugin. Total number of CD45+ cells from each group were used to create viSNE through Cytobank with 2000 iterations, perplexity of 30, and theta of 0.5.
Datta J., Dai X., Bianchi A., De Castro Silva I., Mehra S., Garrido V., Lamichhane P., Singh S., Zhou Z., Dosch A.R., Messaggio F., Ban Y., Umland O., Hosein P.J., Nagathihalli N.S, & Merchant N.B. (2022). Combined MEK and STAT3 inhibition uncovers stromal plasticity by enriching for cancer-associated fibroblasts with mesenchymal stem cell-like features to overcome immunotherapy resistance in pancreatic cancer. Gastroenterology, 163(6), 1593-1612.
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