Icyt synergy cell sorter

Manufactured by Sony

The ICyt Synergy cell sorter is a high-performance flow cytometry instrument designed for efficient cell sorting. It features a compact and modular design that enables flexible configuration to meet the needs of various research applications. The ICyt Synergy provides reliable and accurate cell separation, ensuring the integrity of the sorted cells.

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Lab products found in correlation

Agilent microarray scanner g2565ca, by Agilent Technologies (1 mentions) Mircury lna universal rt microrna pcr, by Qiagen (1 mentions) Feature extraction software v10, by Agilent Technologies (1 mentions) Mirna 107 sep09 scanning protocol, by Agilent Technologies (1 mentions) Rneasy micro kit, by Qiagen (1 mentions) 2100 bioanalyzer, by Agilent Technologies (1 mentions) Rna 6000 pico kit, by Agilent Technologies (1 mentions) Fixation permeabilization reagent, by Thermo Fisher Scientific (1 mentions) Data analysis software, by FlowJo (1 mentions) Cd11c apc cy7, by BioLegend (1 mentions) Brefeldin a solution, by BioLegend (1 mentions) Anti mouse antibodies, by BioLegend (1 mentions) Dynabead, by Thermo Fisher Scientific (1 mentions)

3 protocols using icyt synergy cell sorter

Isolation and Analysis of ILC2 Transcriptome

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ILC2 (CD3⁻CD4⁻Lin⁻ICOS⁺ cells) were isolated from naïve, IL-33 or N. brasiliensis treated mice using fluorescent cell sorting (Sony iCyt Synergy cell sorter) and stored after the addition of TRIzol LS. RNA was extracted using chloroform phase separation, followed by DNase treatment and further purification using Qiagen RNeasy micro kits (performing washes with 100% ethanol to preserve short RNA sequences). RNA integrity was checked using a RNA6000 Pico Kit on an Agilent 2100 Bioanalyzer. Labeling, microarray hybridization, washing, scanning and feature extraction were performed as described previously (22 (link)). As up to 150 mice had to be used to extract sufficient microRNA for microarray analysis of naïve mice, only a single measurement was taken due to cost and ethical considerations. Microarrays were scanned on an Agilent Microarray Scanner (G2565CA) using miRNA_107_Sep09 scanning protocol (Agilent). Spot intensities were then extracted using Agilent's Feature Extraction software v10.7.3. Microarray results were compared to qPCR, obtained using Exiqon's miRCURY LNA Universal RT microRNA PCR Mouse&Rat plates I and II V3.M.

Knolle M.D., Chin S.B., Rana B.M., Englezakis A., Nakagawa R., Fallon P.G., Git A, & McKenzie A.N. (2018). MicroRNA-155 Protects Group 2 Innate Lymphoid Cells From Apoptosis to Promote Type-2 Immunity. Frontiers in Immunology, 9, 2232.

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Multiparametric Flow Cytometry of SVF Cells

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Purified SVF cells were blocked with anti-mouse CD16/32 antibody for 10 minutes on ice. For analysis of immune cell populations, cells were further incubated for 30 minutes on ice in the dark with the following anti-mouse antibodies obtained from BioLegend: APC-CD11b, APC-Cy7-CD11c, PE-CD206 or PE/Cy7-CD206, PerCP/Cy5.5-Ly6G, FITC-CD3, PerCP/Cy5.5-CD19, APC/Cy7-NK1.1. After antibody conjugation, cells were washed with Cell Staining Buffer and analyzed on a Becton-Dickinson LSRII analytical flow cytometer. For analysis of intracellular IL-6, purified SVF cells obtained by pooling the SVF of two AL mice were incubated with 1× Brefeldin A solution (BioLegend) for 4h in culture medium at 37°C. Blocking and cell surface staining were performed as described above followed by fixation and permeabilization using Fixation/Permeabilization Reagents and the protocol recommended by the manufacturer (eBioscience, San Diego, CA). Intracellular staining of IL-6 was performed using anti-mouse PE-IL-6 and negative control (PE-Rat IgG1, κ Isotype control). Cells were analyzed using iCyt Synergy Cell Sorter (Sony Biotechnology San Jose, CA). Analysis of flow cytometry data was performed using FlowJo data analysis software (FlowJo, LLC, Ashland, OR).

Starr M.E., MSteele A., Cohen D.A, & Saito H. (2016). Short-Term Dietary Restriction Rescues Mice From Lethalabdominal Sepsis and Endotoxemia, and Reduces the Inflammatory/Coagulant Potential of Adipose Tissue. Critical care medicine, 44(7), e509-e519.

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Expansion and Transduction of ILC2

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MLN from IL-2/anti-IL-2/IL-25 stimulated mice were collected and pooled. The mouse tissue cell supsension was then depleted of lineage positive cells using biotinylated antibodies (B220, CD3e, CD4, CD5, CD8, CD11b, CD11c, FceRIa, Gr-1, NK 1.1, TER119) and Dynabeads (Thermo Fisher) for magnetic separation. Lineage negative cells were cultured for 24 h with IL-33 (10 ng/ml) and IL-7 (10 ng/ml). 2 × 10⁵ ILC2 were seeded per well and transduced with viral supernatant. The cells were then centrifuged at 900 × g, at 32°C for 90 min and incubated in the presence of IL-33 (10 ng/ml) and IL-7 (10 ng/ml). After 72 h, transduced cells (Live, Lineage⁻ICOS⁺KLRG1⁺mCherry⁺) were sorted using an iCyt Synergy cell sorter (Sony). Isolated ILC2 were cultured with IL-33 (10 ng/ml) and IL-7 (10 ng/ml). Cells were counted every day for 5 days.

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