For primary CD4+ T cells, the percentage of cells that were both GFP+ and mCherry+ was monitored by flow cytometry three and four days after the second infection using a BD LSRFortessa™ X-20 Flow Cytometer (BD Biosciences). J-Lat cells were treated with the indicated concentration of drugs or left untreated. After 18 h, the percentage of GFP+ cells was determined using a MACSQuant VYB FACS analyzer (Miltenyi Biotech GmbH) or FACSCalibur Flow Cytometer (BD Biosciences). Cell viability of primary CD4+ T cells and J-Lat cells was monitored by eBioscience™ fixable viability dye eFluor™ 780 (Invitrogen) and forward-and-side scatter measurement. Analysis was conducted on 3 × 20,000–1Mio. live cells per condition. Data were analyzed using FlowJo 9.5 to 10.8 (Tree Star). All flow cytometry sorting gate strategies are provided in Figure S2 .
Macsquant vyb facs analyzer
Manufactured by Miltenyi Biotec
The MACSQuant VYB FACS analyzer is a flow cytometry instrument designed for cell analysis and sorting. It provides high-performance multicolor detection and precise cell sorting capabilities. The device utilizes advanced optics and fluidics systems to enable accurate and efficient analysis of diverse cell populations.
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3 protocols using macsquant vyb facs analyzer
1
Flow Cytometry Analysis of HIV Latency
2
Flow Cytometry Analysis of HIV Latency
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For primary CD4+ T cells, the percentage of cells that were both GFP+ and mCherry+ was monitored by flow cytometry three and four days after the second infection using a BD LSRFortessa™ X-20 Flow Cytometer (BD Biosciences). J-Lat cells were treated with the indicated concentration of drugs or left untreated. After 18 h, the percentage of GFP+ cells was determined using a MACSQuant VYB FACS analyzer (Miltenyi Biotech GmbH) or FACSCalibur Flow Cytometer (BD Biosciences). Cell viability of primary CD4+ T cells and J-Lat cells was monitored by eBioscience™ fixable viability dye eFluor™ 780 (Invitrogen) and forward-and-side scatter measurement. Analysis was conducted on 3 × 20,000–1Mio. live cells per condition. Data were analyzed using FlowJo 9.5 to 10.8 (Tree Star). All flow cytometry sorting gate strategies are provided in Figure S2 .
3
Multi-color flow cytometry analysis
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Prior to flow cytometry, cells were fixed for 15 min in 4% paraformaldehyde at room temperature. Expression of eGFP/mKO2 or tCD34 was monitored using a MACS Quant VYB FACS analyzer (Miltenyi Biotech GmbH) using a 488 nm, 50 mW DPSS (diode-pumped solid-state) and a 561 nm, 100 mW diode laser respectively and 525/50 nm–586/15 nm band pass filters. A total of at least 30,000 live cells were counted, as determined on the basis of forward scatter channel/side scatter channel (FSC-H/SSC-H) and doublets were excluded based on the FSC-A/FSC-H or SSC-A/SSC-H plot. Single reporter controls where used for compensation purposes. Data were analyzed using third party software (FlowJo).
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