For single-cell FACS analysis, lamina propria lymphocytes and splenocytes were isolated. Surface antigens were stained with a mix of antibodies including a viability marker (Supplementary Table S1 ) and incubated at 4 °C for 20 min. After washing with phosphate-buffered solution (PBS) and centrifugation, all the samples were fixed. For fixation, BD Cytofix/Cytoperm (554722) and BD Perm/Wash (554723) were used following the manufacturer’s instructions. The pellet was then resuspended in PBS. Intracellular antigens were stained with a mix of antibodies (Supplementary Table S1 ). Data were acquired on a FACS LSR II Fortessa 4L (BD) and analyzed with the FlowJo software (version 10.2).
Facs lsr 2 fortessa 4l
Manufactured by BD
The FACS LSR II Fortessa 4L is a flow cytometry instrument designed for research and clinical applications. It features a compact design and is capable of analyzing up to 4 laser beams and 18 fluorescence parameters simultaneously. The core function of the FACS LSR II Fortessa 4L is to perform high-throughput and multi-parameter analysis of cells and other biological particles.
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2 protocols using facs lsr 2 fortessa 4l
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Single-cell FACS Analysis of Immune Cells
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Single-cell FACS Analysis of Immune Cells
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For single-cell FACS analysis, lamina propria lymphocytes and splenocytes were isolated. Surface antigens were stained with a mix of antibodies including a viability marker (Supplementary Table S1 ) and incubated at 4 °C for 20 min. After washing with phosphate-buffered solution (PBS) and centrifugation, all the samples were fixed. For fixation, BD Cytofix/Cytoperm (554722) and BD Perm/Wash (554723) were used following the manufacturer’s instructions. The pellet was then resuspended in PBS. Intracellular antigens were stained with a mix of antibodies (Supplementary Table S1 ). Data were acquired on a FACS LSR II Fortessa 4L (BD) and analyzed with the FlowJo software (version 10.2).
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