Iridium

Manufactured by Standard BioTools

The Iridium is a high-performance mass spectrometer designed for a wide range of analytical applications. It features a compact and modular design, offering flexibility and ease of use. The Iridium provides accurate and reliable mass analysis capabilities, making it a valuable tool for researchers and scientists.

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Lab products found in correlation

Maxpar reagent, by Standard BioTools (2 mentions) Cytobank web server, by Beckman Coulter (2 mentions) Rhodium, by Standard BioTools (2 mentions) Paraformaldehyde (pfa), by Merck Group (2 mentions) Trustain fcx, by BioLegend (1 mentions) Paraformaldehyde (pfa), by Polysciences (1 mentions) Cisplatin, by Abcam (1 mentions) Cytof2 mass cytometer, by Standard BioTools (1 mentions) Eq four element calibration beads, by Standard BioTools (1 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions) Foxp3 fixation permeabilization buffer, by Thermo Fisher Scientific (1 mentions) Foxp3 permeabilization buffer, by Thermo Fisher Scientific (1 mentions) Facs tube, by BD (1 mentions) Helios mass cytometer, by Standard BioTools (1 mentions)

Close spelling variants of this product

Iridium intercalator solution Iridium intercalator Iridium Cell-ID™ Intercalator-Ir Iridium-containing DNA intercalator Iridium interchelator 191/193Ir Iridium intercalator solution Iridium-containing intercalator Iridium 191/193 intercalator Iridium DNA intercalator Cell-ID Iridium intercalator solution

5 protocols using iridium

Mass Cytometry Analysis of CNS Cells

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Purified CNS-single cells (3 × 10⁶ per sample) were first incubated with FcR blocking reagent (cat. 130-092-575; Miltenyi Biotec) according to manufacturer’s protocol and then stained (100-μl final staining reaction volume; 1 h; 4 °C) with a mixture of metal-tagged anti-cytokine/chemokine/growth antibodies (a complete list of antibodies is provided in Additional table 1) conjugated using the MAXPAR reagent (Fluidigm Inc.). Rhodium (1:2,000; Fluidigm Inc.) was added to the cells in the last 20 min of staining. Cells were then washed twice with staining buffer, fixed in 1.6% PFA (Sigma–Aldrich) in PBS (1 h, RT), stained with iridium (1:2,000, 20 min, RT; Fluidigm Inc.), washed again in ultrapure H₂O (to prevent cell loss, the sample was centrifuged at 10,000 g for 1 min), and analyzed on a CyTOF I machine (Fluidigm Inc.), with events acquired at approximately 500 events per second. Internal metal-isotope bead standards were added for sample normalization. Acquired data were uploaded to a Cytobank webserver (Cytobank Inc.) for data processing and for gating out of dead cells and normalization of beads. Rhodium (Rh) and iridium (Ir) (Fluidigm Inc.) inter-chelators were used to identify live/dead cells. At least 20,000 live single cells were analyzed in each sample. The Rh gating was used to assess live/dead cells [29 (link)].

Zilkha-Falb R., Rachutin-Zalogin T., Cleaver L., Gurevich M, & Achiron A. (2020). RAM-589.555 favors neuroprotective and anti-inflammatory profile of CNS-resident glial cells in acute relapse EAE affected mice. Journal of Neuroinflammation, 17, 313.

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CyTOF Staining and Analysis of Fetal Liver Cells

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Total FL cells were collected and incubated with TruStain fcX (Biolegend) for FC blocking. After being washed twice with staining buffer (PBS containing 1% bovine serum albumin and 0.05% sodium azide), the FL cells were stained with a mixture of metal‐tagged antibodies (see Table S1, Supporting Information, for the complete antibodies list). All antibodies were conjugated using the MAXPAR reagent (Fluidigm Inc.). Rhodium (1:2000; Fluidigm Inc.) was added to the cells for the last 20 min of staining. Cells were fixed with 1.6% PFA (Sigma‐Aldrich) in PBS and stained with iridium (Fluidigm Inc.).The samples were analyzed on a CyTOF III machine (Fluidigm Inc.). Acquired data were processed using a Cytobank web server (Cytobank Inc.). Rhodium (Rh) and iridium (Ir) (Fluidigm Inc.) intercalators were used to identify live/dead cells.

Geng X., Zhang C., Li M., Wang J., Ji F., Feng H., Xing M., Li F., Zhang L., Li W., Chen Z., Hickson I.D., Shen H, & Ying S. (2022). PICH Supports Embryonic Hematopoiesis by Suppressing a cGAS‐STING‐Mediated Interferon Response. Advanced Science, 9(7), 2103837.

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Cisplatin-Induced Mass Cytometry Analysis

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Up to 4×10⁶ splenocytes were pulsed with 12.5μM Cisplatin (BioVision) in PBS for 1 min prior to quenching with CyTOF staining media (Mg⁺/Ca⁺ HBSS containing 2% FBS (Multicell), 10mM HEPES (Cornning), and FBS underlay. Cells were then resuspended in staining media containing metal-tagged surface antibodies (Table S1) and Fc block (CD16/32; in house) for 30 min at 4°C. Cells were fixed, permeablized and stained with metal tagged intracellular antibodies (Table S1) using the eBioscience™ Foxp3 / Transcription Factor Staining Buffer Set according to manufacturer’s instructions. All antibody concentrations were used at saturating concentrations previously determined by titration. Cells were then incubated overnight in PBS (Multicell) containing 0.3% (ws/v) saponin, 1.6% (v/v) paraformaldehyde (diluted from 16%; Polysciences Inc) and 50 nM Iridium (Fluidigm). Cells were analyzed on a Helios or CyTOF-2 mass cytometer (Fluidigm). EQ Four Element Calibration Beads (Fluidigm) were used to normalize signal intensity over time and data analysis was performed. P14 T cells were gated on (DNA/Iridium⁺, single event length, Cisplatin⁻, B220^lo NK1.1^lo, TCRβ⁺, CD8a⁺ Thy1.1⁺). t-SNE analyses were performed on the P14 cells (perplexity = 30, theta = 0.5, iterations = 1000, equal sampling).

Snell L.M., MacLeod B.L., Law J.C., Osokine I., Elsaesser H.J., Hezaveh K., Dickson R.J., Gavin M.A., Guidos C.J., McGaha T.L, & Brooks D.G. (2018). CD8+ T cell priming in established chronic viral infection preferentially directs differentiation of memory-like cells for sustained immunity. Immunity, 49(4), 678-694.e5.

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Dissociation and Labeling of Human Islet Cells

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Approximately 20,000 human islet equivalents (IEQs) were dissociated using 0.05% trypsin; trypsin was neutralized by the addition of an equal volume of 100% FBS. Cells were passed through a BD FACS tube with a 40 μm strainer top (BD Biosciences, 352235). Dissociated cells were washed once with PBS containing 10% FBS and once with PBS. Cells were resuspended at a density of 10x10⁶/ml in PBS with cisplatin diluted to 1:4000 and incubated at room temperature for 5 min. Cisplatin labelling was stopped by the addition of a 5x volume of PBS with 10% FBS. The cells were then washed twice with PBS with 10% FBS. Dissociated cells were fixed with FoxP3 Fixation/Permeabilization buffer (eBioscience, 00-5123 and 00-5223) at room temperature (RT) for 2 hours followed by twice washing with FoxP3 permeabilization buffer (eBioscience. 00-8333). Antibody labeling was performed in FoxP3 permeabilization buffer for 8 hours at 4°C at a concentration of up to 2 million cells per 300 μl of antibody cocktail, followed by twice washing with FoxP3 permeabilization buffer. Cells were then incubated with the DNA intercalator Iridium (Fluidigm, 201192A) at a dilution of 1:4000 in Fluidigm Fixation and Permeabilization buffer (Fluidigm, 201067) at RT for 1 hour.

Wang Y.J., Golson M.L., Schug J., Traum D., Liu C., Vivek K., Dorrell C., Naji A., Powers A.C., Chang K.M., Grompe M, & Kaestner K.H. (2016). Single-cell mass cytometry analysis of the human endocrine pancreas. Cell metabolism, 24(4), 616-626.

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Multiparametric Profiling of Tumor-Infiltrating Immune Cells

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CD45⁺ cells were enriched from single cell suspensions from orthotopic PDAC tumors using the positive selection kit and biotin labeled anti-CD45.2 antibody StemCell. Enriched cells were resuspended in Maxpar Staining Buffer, blocked with TruStain Fc blocking buffer for 10 minutes and then resuspended in antibody staining cocktail for 30 Cells were washed twice and incubated in 1 μM Cell_Id cisplatin for 5 minutes. Cisplatin was then quenched by addition of PBS + 5% FBS. Cells were then washed, fixed and permeabilized then barcoded using the Cell-ID multiplex Barcoding kit. Pooled cells were then stained with intracellular antibodies using the permeabilization buffer from Foxp3 transcription factor staining kit. They were then washed twice with perm. Buffer and resuspended in 1.6% paraformaldehyde in PBS containing 0.3% saponin and 125 nM Iridium (Fluidigm, Cat. # 201192A). They were stored in this solution at 4°C until the day of acquisition when they were centrifuged down, washed with Maxpar buffer and resuspended in Maxpar water + EQ beads solution, and analyzed on a Helios Mass Cytometer (Fluidigm) at the Sickkids CYTOF facility.

Hezaveh K., Shinde R.S., Klötgen A., Halaby M.J., Lamorte S., Ciudad M.T., Quevedo R., Neufeld L., Liu Z.Q., Jin R., Grünwald B.T., Foerster E.G., Chaharlangi D., Guo M., Makhijani P., Zhang X., Pugh T.J., Pinto D.M., Co I.L., McGuigan A.P., Jang G.H., Khokha R., Ohashi P.S., O’Kane G.M., Gallinger S., Navarre W.W., Maughan H., Philpott D.J., Brooks D.G, & McGaha T.L. (2022). Tryptophan-derived microbial metabolites activate the aryl hydrocarbon receptor in tumor-associated macrophages to suppress anti-tumor immunity. Immunity, 55(2), 324-340.e8.

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