Following induction, GMDs were stained with 5 µg/mL goat anti-human IgG-PE on ice for 30 minutes. Stained GMDs were washed twice with PBSF and then sorted on a Sony iCyt Synergy SY3200 (excitation 488 nm; emission 585/40 nm) equipped with a 130 micron acrylic nozzle. Backscatter and forward scatter signals were used to exclude free cells and droplet debris. Sorting gates were set to capture the highest 0.05% (high stringency) or 0.5% (low stringency) of GMDs based on PE signal. GMDs were analyzed at >1000 events/second. Flow data was collected using Winlist 3D 7.0, exported as FSC files and analyzed using FlowJo v10. P. pastoris cells from sorted droplets were recovered by outgrowth in YPD media containing 100 μg/mL zeocin.
Icyt synergy sy3200
Manufactured by Sony
The ICyt Synergy SY3200 is a versatile flow cytometry system designed for accurate and reliable cell analysis. It features a compact, streamlined design and supports a range of applications, including immunophenotyping, cell sorting, and fluorescence-activated cell sorting (FACS). The system is equipped with multiple lasers and detectors to enable the simultaneous analysis of multiple parameters for each cell. The ICyt Synergy SY3200 is a valuable tool for researchers and clinicians in the fields of life sciences, immunology, and cell biology.
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Lab products found in correlation
4 protocols using icyt synergy sy3200
1
Flow Cytometric Sorting of Glyco-Engineered Pichia
2
Isolation of Murine Mesenchymal Progenitors
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Hindlimbs from E16.5 mice were homogenized and digested with collagenase. TdTomato+ CXC12-GFP-bright, CD45− CDllb−, Gr1−, B220− cells were sorted using a Sony iCyt Synergy SY3200 cell sorter.
3
Fluorescence-Activated Cell Sorting by Flow Cytometry
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For fluorescence-activated cell sorter (FACS) analysis by flow cytometry, Jurkat cells were suspended in phosphate-buffered saline (PBS) containing 1% FBS (FACS buffer). Dead cells were excluded from all analyses and sorting experiments using propidium iodide (PI). Acquisition was carried out on the fluorescein isothiocyanate (FITC) channel for GFP and on the PE channel for PI. Cell fluorescence was assessed using a BD LSR Fortessa instrument (BD Biosciences), and data were analyzed using FlowJo software, version 10.6 (FlowJo, LLC, Ashland, OR). Infected cells were sorted into GFP+ and GFP− subpopulations by flow cytometry using a FACSAria II (BD Biosciences, Franklin Lakes, NJ) or an iCyt Synergy SY3200 (Sony Biotechnology, San Jose, CA) cell sorter at the flow cytometry core of the University of Michigan.
4
FACS Analysis of Jurkat Cells
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For Fluorescence Activated Cell Sorting (FACS) analysis by flow cytometry, Jurkat cells were suspended in phosphate buffered saline (PBS) containing 1% FBS (FACS buffer). Dead cells were excluded from all analyses and sorting experiments using propidium iodide (PI). Acquisition was carried out on the FITC channel for GFP and PE channel for PI. Cell fluorescence was assessed using BD LRS Fortessa (BD Biosciences) and data were analyzed using FlowJo software, version 10.6 (FlowJo, LLC., Ashland, Oregon). Infected cells were sorted into GFP+ and GFP-subpopulations by flow cytometry using FACS Aria II (BD Biosciences, Franklin Lakes, NJ) or iCyt Synergy SY3200 (Sony Biotechnology, San Jose, CA) cell sorters at the flow cytometry core of the University of Michigan.
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