Facs verse flow cytometer

Manufactured by Tree Star

Sourced in United States

The FACS Verse flow cytometer is a device used for the analysis and sorting of cells and other microscopic particles. It utilizes laser technology to detect and measure the physical and chemical characteristics of individual cells or particles as they pass through a fluid stream. The core function of the FACS Verse is to provide high-performance cell analysis and sorting capabilities for research and diagnostic applications.

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Lab products found in correlation

Anti mouse cd11c, by BD (1 mentions) Cd8 antibody, by BD (1 mentions) Flowjov software, by Tree Star (1 mentions) Anti ifn γ, by BD (1 mentions) Cytofix cytoperm solution, by BD (1 mentions) Mhcii, by BD (1 mentions) Anti il 4, by BD (1 mentions) Cd11b, by BioLegend (1 mentions) Golgiplug, by BD (1 mentions) Anti cd11c apc clone b ly6, by BD (1 mentions) Pancoll human, by PAN Biotech (1 mentions) Stereomicroscope, by Leica (1 mentions) Ly6g pecy7, by Thermo Fisher Scientific (1 mentions) Cd19 apc cy7, by Thermo Fisher Scientific (1 mentions) Cd11b pe cy7 m1 70, by Thermo Fisher Scientific (1 mentions) Dnase 1, by Roche (1 mentions) Collagenase d, by Roche (1 mentions)

4 protocols using facs verse flow cytometer

Analyzing Mouse Immune Cell Phenotypes

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Mouse BMDCs or splenocytes were washed with PBS. For surface marker extracellular staining, the cells were incubated with anti-mouse CD11c (557,400, BD Biosciences), CD80 (560,526, BD Biosciences), CD86 (552,692, BD Biosciences), MHC-II (562,367, BD Biosciences), CD11b (101,211, Biolegend), CD4 (553,046, BD Biosciences), or CD8 antibodies (551,162, BD Biosciences) at 4 °C for 30 min. For intracellular cytokine staining, the splenocytes were stimulated with DnaJ (10 μg/ml) for 8 h in presence of Golgi plug™ (51-2301KZ, BD Bioscience). The splenocytes were then treated for surface markers (CD4 or CD8), fixed/permeabilised with a Cytofix/Cytoperm solution (51-2090KZ, BD Bioscience) and then stained with anti-IFN-γ (557,735, BD Biosciences) and anti-IL-4 (554,435, BD Biosciences) antibodies at 20–25 °C for 30 min. All events were acquired on a FACSverse flow cytometer and analysed using the FlowJoV software (Tree Star).
To determine cytokine (IFN-γ, TNF-α, IL-1β, IL-10, IL-17a, MCP-1, IL-1α, and IL-6) concentrations in the uterine tissue, multi-analyte flow assay kits (740,446, Biolegend, San Diego, CA, USA) were used as indicated by the manufacturer.

Guo F., Tang Y., Zhang W., Yuan H., Xiang J., Teng W., Lei A., Li R, & Dai G. (2022). DnaJ, a promising vaccine candidate against Ureaplasma urealyticum infection. Applied Microbiology and Biotechnology, 106(22), 7643-7659.

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Phenotypic Profiling of Monocyte Subsets

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Surface antigens were studied on peripheral blood mononuclear cells (PBMC) as previously described20 (link), 21 (link). Briefly, PBMCs were isolated by gradient centrifugation using Lymphocyte Separation Medium LSM 1077 (PAA Laboratories GmbH, Austria) from EDTA-treated blood. PBMC were further suspended in phosphate buffer saline (PBS) containing 1% heat inactivated fetal bovine serum (FBS) (Gibco, Life Technologies, USA) and were used immediately after isolation. 250000 PBMCs were stained for 20 minutes with fluorochrome-conjugated monoclonal antibodies: anti-CD14-PerCP (MΦP9), anti-CD16-APC-H7 (3G8), anti-HLA-DR-PE-Cy7 (L243) (BD, Pharmingen, CA, USA). After staining, cells were washed twice with PBS containing 1% FBS.
Cells were processed in the FACS Verse flow cytometer and analyzed using FlowJo software (TreeStar, USA). Monocytes were gated according to FSC (forward scatter) and SSC (side scatter) signals as described previously21 (link). Subsequently, cells were gated in a HLA-DR/CD14 plot to exclude HLA-DR negative natural killer cells. Finally, we analyzed cells for CD14 and CD16 expression, which allowed for discrimination of major monocyte subpopulations as described23 (link). Absolute monocyte number/mm³ was calculated20 (link).

Mikołajczyk T.P., Osmenda G., Batko B., Wilk G., Krezelok M., Skiba D., Sliwa T., Pryjma J.R, & Guzik T.J. (2015). Heterogeneity of peripheral blood monocytes, endothelial dysfunction and subclinical atherosclerosis in patients with Systemic Lupus Erythematosus. Lupus, 25(1), 18-27.

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Monocyte Subpopulation Characterization

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Surface antigens were studied in peripheral blood mononuclear cells (PBMCs) as previously described [36 (link)]. Briefly, PBMCs were isolated by gradient centrifugation using Pancoll human (PAN-Biotech GmbH, Aidenbach, Germany) from the ethylenediaminetetraacetic acid- (EDTA-) treated blood. PBMCs were further suspended in phosphate-buffered saline (PBS) containing 1% heat-inactivated fetal bovine serum (FBS) (Gibco, Life Technologies, USA) and were used immediately after isolation. A total of 500,000 PBMCs were stained for 20 minutes with fluorochrome-conjugated monoclonal antibodies: anti-CD14-APC-H7 (clone MΦP9), anti-CD16-PE (clone 3G8), anti-human leukocyte antigen (HLA)-DR-PE-Cy7 (L243), anti-CD11b/Mac-1-Pacific Blue (clone ICRF44), anti-CD11c-APC (clone B-LY6), and anti-CD45RA-FITC (clone L48) (BD, Pharmingen, CA, USA). After staining, cells were washed twice with PBS containing 1% FBS. Cells were processed in the FACS Verse flow cytometer and analyzed using FlowJo software (Tree Star, USA). Monocytes were gated according to forward scatter (FSC) and side scatter (SSC) signals as described previously. Subsequently, cells were gated in an HLA-DR/CD14 plot to exclude HLA-DR-negative Natural Killer cells. Finally, we analyzed cells for CD14 and CD16 expression, which allowed for discrimination of major monocyte subpopulations: CD14++CD16−, CD14++CD16+, and CD14+CD16++.

Maga P., Mikolajczyk T.P., Partyka L., Krzanowski M., Malinowski K.P, & Nizankowski R. (2016). Percutaneous Transluminal Angioplasty in Patients with Peripheral Arterial Disease Does Not Affect Circulating Monocyte Subpopulations. BioMed Research International, 2016, 2708957.

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Flow Cytometry Analysis of Mouse Brain Immune Cells

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After perfusion with saline, the ChP was removed under a stereomicroscope (Leica) and collected in 200 µl Dulbecco's Modified Eagle Medium (DMEM) + 10% fetal calf serum (FCS). ChP cells were isolated by incubating the samples in 2 ml of digestion mix [DMEM + 10% FCS + 0.4% DNASEI (#11284932001, Roche) + 3% CollagenaseD (#11088866001, Roche)], 10 min at 37 °C, and then mechanically dissociated. Brain homogenates of both hemispheres were prepared by the same dissociation/digestion protocol. Cerebral mononuclear cells were subsequently isolated using a 70 and 40% discontinuous Ficoll gradient. The following mouse antigen-specific antibodies were purchased from eBioscience: CD45 eF450 (30-F11), CD11b PE-Cy7 (M1/70), CD3 V510 or FITC (17A2), CD19 APC-Cy7 (eBisID3), CD8 PE (53-6.7), CD4 PerCP5.5 (45-0042-82), Ly6C APC (HK1.4), Ly6G PE Cy7 (RB6-8C5) and Ly6G PE (1A8). To quantify the various cell populations, cells were stained with specific antibodies in accordance with the manufacturer's protocols, acquired in a FACSVerse flow cytometer and analyzed with FlowJo software (version 10, Tree Star).

Llovera G., Benakis C., Enzmann G., Cai R., Arzberger T., Ghasemigharagoz A., Mao X., Malik R., Lazarevic I., Liebscher S., Ertürk A., Meissner L., Vivien D., Haffner C., Plesnila N., Montaner J., Engelhardt B, & Liesz A. (2017). The choroid plexus is a key cerebral invasion route for T cells after stroke. Acta neuropathologica, 134(6).

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