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Lsr 2 digital flow cytometer

Manufactured by BD
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The BD LSR II digital flow cytometer is an instrument designed for the detection and analysis of cells and particles in a fluid sample. It utilizes laser technology to measure and characterize the physical and fluorescent properties of individual cells or particles as they pass through the instrument's flow cell. The LSR II provides high-performance detection and data acquisition capabilities for a wide range of applications in fields such as immunology, cell biology, and disease research.

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Lab products found in correlation

Collagenase type xi, by Merck Group (2 mentions) Perm wash, by BD (2 mentions) Fix perm solution, by BD (2 mentions) Type 4 bovine pancreatic dnase, by Merck Group (2 mentions) Granzyme b clone gb12, by Thermo Fisher Scientific (2 mentions) Cd69 clone h1.2f3, by BD (2 mentions) Ifnγ clone xmg1, by BD (2 mentions) Golgistop, by BD (2 mentions) Live dead fixable dead cell stain kit, by Thermo Fisher Scientific (2 mentions) Facsdiva software, by BD (2 mentions) Gentlemacs tissue dissociator, by Miltenyi Biotec (2 mentions) Anti mouse cd16 cd32 fc block, by BD (2 mentions) Anti ifn γ and anti tnfα, by BioLegend (1 mentions) Anti gl7, by BioLegend (1 mentions) 7 aminoactinomycin d, by BioLegend (1 mentions) Anti cd19, by BioLegend (1 mentions)

5 protocols using lsr 2 digital flow cytometer

1

Multiparametric Immune Cell Profiling

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Spleens from infected animals were dissociated, and single-cell suspensions were stained with the following antibodies from BioLegend: anti-GL7, anti-CD138, anti-CD19, anti-CD45.1, anti-CD8, anti-CD4, anti-Thy1.1, anti-Thy1.2, anti-CD11c, anti-CD11b, and anti-IgM. 7-Aminoactinomycin D and counting beads (Flow Cytometry Absolute Count Standard) were added to each sample before acquisition to allow calculation of cells per sample. Intracellular cytokine staining was performed by standard methods, staining intracellularly with anti–IFN-γ and anti-TNFα (BioLegend). Intracellular granzyme B and LCMV were detected using anti–granzyme B or anti-LCMV antibodies, respectively. Samples were acquired using an LSR II digital flow cytometer (BD Biosciences), and data were analyzed using FlowJo software version 10.0.7 (Tree Star).
Moseman E.A., Wu T., de la Torre J.C., Schwartzberg P.L, & McGavern D.B. (2016). Type I interferon suppresses virus-specific B cell responses by modulating CD8+ T cell differentiation. Science immunology, 1(4), eaah3565.
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2

Murine Myeloid Cell Profiling

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Blood was collected from B6 mice in heparinized capillary tubes and placed into FACS buffer (PBS plus 2% FBS). Red blood cells were lysed by addition of ACK lysis buffer and were incubated for 5–10 min at RT. Cells were pelleted and stained in FACS buffer with CD11b PE/Cy7 (M1/70), Ly6C Alexa Fluor 488 or Brilliant Violet 570 (HK1.4), Ly6G Pacific Blue (1A8), CD115 APC (AFS98). All directly conjugated antibodies purchased from BioLegend. Samples were acquired using an LSR II digital flow cytometer (BD), and data were analyzed using FlowJo software version 10.0.7 (Tree Star).
Russo M.V., Latour L.L, & McGavern D.B. (2018). Distinct myeloid cell subsets promote meningeal remodeling and vascular repair after mild traumatic brain injury. Nature immunology, 19(5), 442-452.
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3

Flow Cytometry Analysis of Basophils

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We used a LSRII digital flow cytometer (BD Biosciences) equipped with four lasers (535, 488, 633 and 405 nm), two light-scatter detectors (yielding forward and side scatter data) and 18 fluorescent detectors. Analytical gates for basophils included ⩾300 cells [26] .
Gernez Y., Waters J., Mirković B., Lavelle G.M., Dunn C.E., Davies Z.A., Everson C., Tirouvanziam R., Silver E., Wallenstein S., Chotirmall S.H., McElvaney N.G., Herzenberg L.A, & Moss R.B. (2016). Blood basophil activation is a reliable biomarker of allergic bronchopulmonary aspergillosis in cystic fibrosis. The European respiratory journal, 47(1).
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4

Lung dissociation and immune cell analysis

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The left lung lobe was excised post BAL, crudely dissociated using the GentleMACS™ tissue dissociator (Miltenyi Biotech, Germany) and digested upon incubation at 37°C in buffer containing 1 mg/mL collagenase Type XI and 80 units/mL Bovine Pancreatic DNase Type IV (both Sigma-Aldrich, Dorset, UK). For intracellular cytokine staining (ICS) cells were stimulated with PMA (50 ng/mL) and ionomycin (500 ng/mL) with BD GolgiStop™ (BD Biosciences) for 3 h at 37°C. Lung and BAL cells were incubated with anti-mouse CD16/CD32 (FC Block, BD Biosciences) prior to staining for cell surface markers: CD3e (clone 500A2), CD4 (clone RM4-5), CD8a (clone 53-6.7), NK1.1 (clone PK136), NKp46 (clone 29A1.4) and CD69 (clone H1.2F3) (all BD Biosciences). Cells were washed and stained with Live/Dead fixable dead cell stain kit (Invitrogen), followed by incubation with BD Fix/Perm solution (BD Biosciences). For ICS, cells were stained for IFN-γ (clone XMG1.2, BD Biosciences) and granzyme B (clone GB12, Invitrogen) in BD PermWash™ (BD Biosciences). Data were acquired using a BD LSR II digital flow cytometer (BD Biosciences) and BD FACS Diva software. Analysis was performed using FlowJo 9.3.1.2 software.
Jayaraman A., Jackson D.J., Message S.D., Pearson R.M., Aniscenko J., Caramori G., Mallia P., Papi A., Shamji B., Edwards M., Westwick J., Hansel T., Stanciu L.A., Johnston S.L, & Bartlett N.W. (2014). IL-15 complexes induce NK cell and T cell responses independent of type I IFN signalling during rhinovirus infection. Mucosal immunology, 7(5), 1151-1164.
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5

Lung dissociation and immune cell analysis

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The left lung lobe was excised post BAL, crudely dissociated using the GentleMACS™ tissue dissociator (Miltenyi Biotech, Germany) and digested upon incubation at 37°C in buffer containing 1 mg/mL collagenase Type XI and 80 units/mL Bovine Pancreatic DNase Type IV (both Sigma-Aldrich, Dorset, UK). For intracellular cytokine staining (ICS) cells were stimulated with PMA (50 ng/mL) and ionomycin (500 ng/mL) with BD GolgiStop™ (BD Biosciences) for 3 h at 37°C. Lung and BAL cells were incubated with anti-mouse CD16/CD32 (FC Block, BD Biosciences) prior to staining for cell surface markers: CD3e (clone 500A2), CD4 (clone RM4-5), CD8a (clone 53-6.7), NK1.1 (clone PK136), NKp46 (clone 29A1.4) and CD69 (clone H1.2F3) (all BD Biosciences). Cells were washed and stained with Live/Dead fixable dead cell stain kit (Invitrogen), followed by incubation with BD Fix/Perm solution (BD Biosciences). For ICS, cells were stained for IFN-γ (clone XMG1.2, BD Biosciences) and granzyme B (clone GB12, Invitrogen) in BD PermWash™ (BD Biosciences). Data were acquired using a BD LSR II digital flow cytometer (BD Biosciences) and BD FACS Diva software. Analysis was performed using FlowJo 9.3.1.2 software.
Jayaraman A., Jackson D.J., Message S.D., Pearson R.M., Aniscenko J., Caramori G., Mallia P., Papi A., Shamji B., Edwards M., Westwick J., Hansel T., Stanciu L.A., Johnston S.L, & Bartlett N.W. (2014). IL-15 complexes induce NK cell and T cell responses independent of type I IFN signalling during rhinovirus infection. Mucosal immunology, 7(5), 1151-1164.
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