Lrsfortessa flow cytometer

Manufactured by BD

Sourced in United States

The LRSFortessa flow cytometer is a laboratory instrument designed for cell analysis. It is capable of detecting and measuring multiple parameters of individual cells or particles within a sample. The LRSFortessa utilizes laser light sources and detectors to analyze the properties of cells, such as size, granularity, and fluorescence. This data can be used to identify and characterize different cell types or populations within a heterogeneous sample.

Automatically generated - may contain errors

Lab products found in correlation

17 protocols using lrsfortessa flow cytometer

Hoechst 33342 Side Population Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

For side population studies, 1 million cells were incubated with Hoechst 33342 (5 μg/ml) for 90 min at 37°C with a shake every 10 min. Cells incubated with the addition of verapamil (50 μM) were used as a control. Side population cells were analyzed on BD LRSFortessa flow cytometer.

Zeng D., Liu M, & Pan J. (2016). Blocking EZH2 methylation transferase activity by GSK126 decreases stem cell-like myeloma cells. Oncotarget, 8(2), 3396-3411.

+ Open protocol

+ Expand

Annexin V Apoptosis Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

1 × 106 cells/ml were harvested and resuspended in 1 × annexin-binding buffer and incubated with 5 μl of Annexin V–488 (Alexa Fluor 488 Annexin V/Dead Cell Apoptosis Kit; Life technologies, Carlsbad, CA, United States) for 15 min at room temperature. Cells were then resuspended in PBS and measured with a BD LRSFortessa Flow cytometer. Flow cytometric data were analyzed with FCSalyzer 0.9.15-alpha software.

Granata A., Kasioulis I., Serrano F., Cooper J.D., Traylor M., Sinha S, & Markus H.S. (2022). The Histone Deacetylase 9 Stroke-Risk Variant Promotes Apoptosis and Inflammation in a Human iPSC-Derived Smooth Muscle Cells Model. Frontiers in Cardiovascular Medicine, 9, 849664.

+ Open protocol

+ Expand

Bacterial Membrane Staining and Analysis

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Control or AEA-treated bacteria were exposed to 1 µg/mL of diamidino-2-phenylindole (DAPI) for 30 min together with 10 µg/mL Nile Red (APExBIO, Boston, MA, USA) for 30 min at 37 °C. After incubation, the samples were analyzed by flow cytometry (LRS-Fortessa flow cytometer, BD Biosciences). The 355 nm laser was used for DAPI, and the fluorescence was collected by the blue (450 nm) filter. DAPI is a blue-fluorescent probe that fluoresces brightly upon selectively binding to DNA [11 (link)]. The Nile Red was analyzed using the 561 nm yellow–green laser excitation, and data were collected using the 635 nm filter [20 (link)].

Wolfson G., Sionov R.V., Smoum R., Korem M., Polacheck I, & Steinberg D. (2023). Anti-Bacterial and Anti-Biofilm Activities of Anandamide against the Cariogenic Streptococcus mutans. International Journal of Molecular Sciences, 24(7), 6177.

+ Open protocol

+ Expand

Multiparametric Flow Cytometry for DC Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols

For DCs flow cytometry, PBMCs were centrifuged, pelleted and washed with Phosphate-buffered saline (PBS) and stained for 35 min at room temperature with LIVE/DEAD Fixable Aqua Dead Cell Stain (Life Technologies), BV421 CD86, BV650 CD11c, BV711 HLA-DR, BV786 CCR7 (CD197), FITC Lin-2 (CD3, CD14, CD19, CD20, CD56), BV605 CD16, PeCF594 PD-L1 (CD274), APC Integrin-β7 (BD Biosciences), PerCPCy5,5 CD4, APCCy7 CD1c, PeCy7 CD141 (BioLegend) and AF700 CD123 (R&D, San Diego, CA) antibodies. Then PBMCs were washed with Permeabilization Buffer 10X diluted 1:10 (eBioscience™), permeabilized by Fixation/Perm buffer (eBioscience™), and intracellularly stained with PE IDO (eBioscience, San Diego, CA, USA) antibody. DCs were gated based on Lin-2 HLA-DR expression. Each subset (mDCs and pDCS) was gated based on CD123 and CD11c expression. mDCs subsets were gated by using CD16, CD1c and CD141 staining, for gating strategy see Supplementary Fig. 1. Flow cytometry analyses were performed on an LRS Fortessa flow cytometer using FACS Diva software (BD Biosciences). Data were analyzed using the FlowJo software (Treestar, Ashland, OR). At least 1 × 10⁶ events were acquired per sample.

Pérez-Gómez A., Vitallé J., Gasca-Capote C., Gutierrez-Valencia A., Trujillo-Rodriguez M., Serna-Gallego A., Muñoz-Muela E., Jiménez-Leon M.D., Rafii-El-Idrissi Benhnia M., Rivas-Jeremias I., Sotomayor C., Roca-Oporto C., Espinosa N., Infante-Domínguez C., Crespo-Rivas J.C., Fernández-Villar A., Pérez-González A., López-Cortés L.F., Poveda E, & Ruiz-Mateos E. (2021). Dendritic cell deficiencies persist seven months after SARS-CoV-2 infection. Cellular and Molecular Immunology, 18(9), 2128-2139.

+ Open protocol

+ Expand

Isolation and Characterization of Testis Interstitial Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Single-cell suspensions from the interstitial space of rat testis were obtained by decapsulating one testis per rat in 3 ml of ice-cold PBS. After removal of the tunica albuginea, 1 ml of DMEM + 1% collagenase was added and seminiferous tubules were gently teased apart using fine forceps. Seminiferous tubules were then rinsed with 1 ml of PBS, snap frozen and stored at −80 °C until use. Interstitial cells were centrifuged (5 min, 1200 rpm, 4°C Beckman table top), filtered through a 70 μm cell strainer, resuspended in 1 ml of PBS, and adjusted to approximately 1 × 10⁶ cells/ml. Cells were centrifuged (1200 rpm at 4°C) for 5 min, resuspended in 200 μl of FACS buffer (PBS + 2% inactivated bovine serum) with anti-CD11b-APC (1:200; eBiosinces, San Diego, CA). Cells were incubated on ice in the dark for 30 min, then washed with 3 ml of FACS buffer, centrifuged at 1200 rpm for 5 min at 4°C and resuspended in 0.5 ml FACS buffer containing propidium iodide (2 μg/ml). For each sample, 50,000 cells were analyzed on a BD LRSFortessa flow cytometer with FACSDiva software and interpreted with FlowJo software. Additionally, single-cell suspensions from lymph nodes were reacted with or without antibodies to set gates, compensation, and autofluorescence. Only live cells were used for analysis. The total numbers of CD11b+ cells (leukocytes) were determined for each rodent based on cell counts.

Stermer A.R., Murphy C.J., Ghaffari R., Di Bona K.R., Voss J.J, & Richburg J.H. (2017). Mono-(2-ethylhexyl) phthalate-induced Sertoli cell injury stimulates the production of pro-inflammatory cytokines in Fisher 344 rats. Reproductive toxicology (Elmsford, N.Y.), 69, 150-158.

+ Open protocol

+ Expand

Apoptosis Quantification by Flow

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were harvested, washed twice with PBS and stained with an annexin V-FITC/PI apoptosis detection kit. The stained cells were analyzed by a BD LRS Fortessa flow cytometer.

Zhang J.Y., Zhou B., Sun R.Y., Ai Y.L., Cheng K., Li F.N., Wang B.R., Liu F.J., Jiang Z.H., Wang W.J., Zhou D., Chen H.Z, & Wu Q. (2021). The metabolite α-KG induces GSDMC-dependent pyroptosis through death receptor 6-activated caspase-8. Cell Research, 31(9), 980-997.

+ Open protocol

+ Expand

Immunophenotyping of Mesenchymal Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For immunophenotyping, MSC lines were passaged and centrifuged and the pellet was resuspended in PBS. A total of 5 × 10⁵ cells was used for labeling with the following antibodies in the concentration 1/50: Sca1PE-Cy7 (BD Biosciences, San Jose, CA, USA), CD45-PerCP (eBioscience, San Diego, CA, USA), CD44-PE (BD Bioscience), CD90-APC (BD Bioscience), CD34-AlexaFluor647 (BD Bioscience), and control isotypes. Cells were incubated in 100 μL of binding buffer (ThermoFisher Scientific) with annexin-V-FITC and 7-AAD (BD Biosciences, San Jose, CA, USA) for 15 minutes in the dark at RT. After the incubation period, cells were washed twice with PBS, and the data acquisition and analysis were performed using a LRSFortessa flow cytometer (BD Biosciences). At least 10,000 events were acquired and analyzed.

Souza B.S., da Silva K.N., Silva D.N., Rocha V.P., Paredes B.D., Azevedo C.M., Nonaka C.K., Carvalho G.B., Vasconcelos J.F., dos Santos R.R, & Soares M.B. (2017). Galectin-3 Knockdown Impairs Survival, Migration, and Immunomodulatory Actions of Mesenchymal Stromal Cells in a Mouse Model of Chagas Disease Cardiomyopathy. Stem Cells International, 2017, 3282656.

+ Open protocol

+ Expand

Anionic Dextran Staining of S. mutans

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Overnight cultures of S. mutans were diluted in fresh BHI media to an OD of 0.2. Then, the cells were incubated in the absence or presence of various concentrations of AEA (1.56–50 µg/mL) and 0.05% ethanol for 2 h in fresh BHI media at 37 °C in 95% air/5% CO₂. After incubation, the samples were stained with 10 µg/mL Alexafluor⁶⁴⁷ -conjugated anionic Dextran 10,000 (Invitrogen Live Technologies Corporation, Eugene, OR, USA) for 20 min at 37 °C in 95% air/5% CO₂. Fluorescence intensity was analyzed by flow cytometry (LRS-Fortessa flow cytometer, BD Biosciences, San Jose, CA, USA) using the 640 nm laser excitation, and the fluorescence was collected using the 670 nm filter [13 (link)].

+ Open protocol

+ Expand

Lung Immune Cell Profiling Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

To determine immune cell populations, lungs were digested (digestion buffer: 7 mL/sample Dulbecco’s modified Eagle’s medium + 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, 2% bovine serum albumin, 0.03 mg/mL Liberase Blendzyme 3 Collagenase Cocktail, 50 U/mL DNAse) and immune cells were isolated. Cells were stimulated for 4 hours with phorbol 12-myristate 13-acetate (50 ng/mL; Sigma-Aldrich), Ionomycin (1 μg/mL; Calbiochem), and brefeldin A. Cells were subsequently labeled with monoclonal antibodies. Specifically, Live/dead (Zombie UV Dye; BioLegend, San Diego, CA, USA), B220 – BV605 (clone RA3–6B2, BioLegend), CD45 – PEDazzle (clone 104, BioLegend), CD11b – eFluor450 (clone M1/70, eBioscience, San Diego, CA, USA), CD11c – BV711 (clone N418, BioLegend), NK1.1 – FITC (clone PK136, BioLegend), F4/80 – AF700 (clone BM8, eBioscience), Gr1 – APC (clone RB6–8C5, eBioscience), CD8 – BV510 (clone 53–6.7, BioLegend), and IL6 – PE (clone MP5–20F3, eBioscience). Data were collected using an LRS Fortessa flow cytometer (BD Biosciences) and analyzed by FlowJo X software (vX10, Tree Star, Ashland, OR, USA).

Alarcon P.C., Damen M.S., Ulanowicz C.J., Sawada K., Oates J.R., Toth A., Wayland J.L., Chung H., Stankiewicz T.E., Moreno-Fernandez M.E., Szabo S., Zacharias W.J, & Divanovic S. (2023). Obesity amplifies influenza virus-driven disease severity in male and female mice. Mucosal immunology, 16(6), 843-858.

+ Open protocol

+ Expand

CFSE Staining and HIV Peptide Stimulation

Check if the same lab product or an alternative is used in the 5 most similar protocols

PBMCs, 1 × 10⁶ cells/mL, were stained at 37°C for 20 minutes with 0.5 μM CellTrace carboxyfluorescein succinimidyl ester (CFSE; Thermo Fisher Scientific) according to the manufacturer’s protocol. Cells were washed twice with RPMI-1640 medium (RPMI) supplemented with 10% FBS (Sigma-Aldrich), and plated in 96-well round-bottom polystyrene plates, 200 μL per well. Experimental samples were incubated with 20 ng/mL of an overlapped HIV (Gag)-specific peptide pool (NIH AIDS Reagent Program). Positive control well was stimulated with 2 μg/mL of SEB (Sigma-Aldrich) and the negative control contained unstimulated PBMCs. Subsequently, cells were cultured for 5 days in R-10 medium (RPMI supplemented with 10% FBS, 100 U/mL penicillin G, 100 μL/mL streptomycin sulfate [Thermo Fisher Scientific], 1.7 mM sodium L-glutamine [Lonza] and 50 IU/mL IL-2 [R&D Systems]). On day 5, cells were collected and stained for viability using Violet LIVE/DEAD Cell Stain kit (Invitrogen) and anti-CD3-APC-H7 (clone SK7; BD Biosciences) and anti-CD8-PE (clone RPA-T8; Biolegend). Finally, cells were washed and fixed for 20 minutes at 4°C with 4% paraformaldehyde solution (PFA; Sigma-Aldrich). Multiparametric flow cytometry analyses were performed on an LRS Fortessa flow cytometer using FACS Diva software (BD Biosciences). Data were analyzed using the FlowJo 10.7.1 software (Treestar).

Gasca-Capote C., Lian X., Gao C., Roseto I.C., Jiménez-León M.R., Gladkov G., Camacho-Sojo M.I., Pérez-Gómez A., Gallego I., Lopez-Cortes L.E., Bachiller S., Vitalle J., Rafii-El-Idrissi Benhnia M., Ostos F.J., Collado-Romacho A.R., Santos J., Palacios R., Gomez-Ayerbe C., Muñoz-Medina L., Ruiz-Sancho A., Frias M., Rivero-Juarez A., Roca-Oporto C., Hidalgo-Tenorio C., Rull A., Olalla J., Lopez-Ruz M.A., Vidal F., Viladés C., Mastrangelo A., Cavassini M., Espinosa N., Perreau M., Peraire J., Rivero A., López-Cortes L.F., Lichterfeld M., Yu X.G, & Ruiz-Mateos E. (2024). The HIV-1 reservoir landscape in persistent elite controllers and transient elite controllers. The Journal of Clinical Investigation, 134(8), e174215.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!