Log In Sign up
The largest database of trusted experimental protocols

5 laser lsr fortessa

Manufactured by BD
Visit Supplier
Sourced in United States

The BD 5-laser LSR Fortessa is a flow cytometry instrument that utilizes five lasers to analyze cells and particles. It is designed to provide high-performance, multi-parameter data acquisition and analysis.

Automatically generated - may contain errors

Lab products found in correlation

Abc total antibody compensation bead kit, by Thermo Fisher Scientific (2 mentions) Trustain fcx, by BioLegend (1 mentions) Arc amine reactive compensation bead kit, by Thermo Fisher Scientific (1 mentions) Brilliant stain buffer, by BD (1 mentions) Live dead fixable aqua dead cell stain, by BD (1 mentions) Macs comp bead kit anti rea, by Miltenyi Biotec (1 mentions) Fluorofix buffer, by BioLegend (1 mentions) Facs lysing solution, by BD (1 mentions) Trucount tube, by BD (1 mentions) Cd11b fitc, by BioLegend (1 mentions) Cd11c pe cy7, by BioLegend (1 mentions) Cd11b apc, by BioLegend (1 mentions) Cd19 percp cy5, by BioLegend (1 mentions) Siglech apc, by BioLegend (1 mentions) B220 fitc, by BD (1 mentions) Mhcii bv510, by BioLegend (1 mentions) Cd8α bv605, by BioLegend (1 mentions) Ly6c bv650, by BioLegend (1 mentions) Cd3 bv785, by BioLegend (1 mentions) Kaluza v1, by Beckman Coulter (1 mentions)

Spelling variants of this product

Fortessa (1232 citations) Fortessa LSR (45 citations)

5 protocols using 5 laser lsr fortessa

1

Intracellular Signaling in Splenocytes

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Splenocytes from both immunized and unimmunized animals were harvested as described above. From each animal, four replicates of five million splenocyte cultures were grown for 20 minutes at 37°C in 5 mL of serum-free (SF) culture medium (RPMI-1640, phenol red free, 1% glutamine and 5% penicillin–streptomycin) or SF containing freshly 400ng/ml IL6-sIL6Rα (= Hyper-IL6; Fischer et al., 1997 (link)) (9038-SR-025, R&D Systems). After this culture period, the cells were pelleted, fixed and then permeabilized to enable intracellular staining using pY705 STAT3-PE (BioLegend) along with extracellular markers [B220-FITC, MHCII-BV510, CD138-BV421, CD3-BV786 and gp130-APC], according to the True-Phos™ Protocol provided by Biolegend (Xu et al., 2017 (link)). The samples were assayed on a 5 laser LSR Fortessa (BD Biosciences) and data was analyzed using FlowJo software version 9.3.2 (TreeStar) as described above.
Yousif A.S., Ronsard L., Shah P., Omatsu T., Sangesland M., Bracamonte Moreno T., Lam E.C., Vrbanac V.D., Balazs A.B., Reinecker H.C, & Lingwood D. (2020). The persistence of interleukin-6 is regulated by a blood buffer system derived from dendritic cells. Immunity, 54(2), 235-246.e5.
+ Open protocol
+ Expand
2

Dermal Fibroblast Phenotyping by Flow Cytometry

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Dermal fibroblasts at a low passage were harvested using Accutase™ following the manufacturer instructions and washed in FACS wash Buffer (composition: 1x PBS without calcium and magnesium, 0.5% BSA, 2mM EDTA and 2mM Sodium Azide). Cells were then blocked with TruStain FcX (BioLegend) and subsequently stained with 2 panels of antibodies and matching isotype controls (Table S3) with brilliant stain buffer (BD Biosciences) and LIVE/DEAD™ Fixable Aqua – Dead cell stain in polypropylene tubes at 4°C. Cells were washed in FACS wash Buffer and fixed using FluoroFix Buffer (BioLegend). Samples were acquired on a 5-laser LSR Fortessa (BD Biosciences) with single color compensation controls (AbC™ Total Antibody Compensation Bead Kit and ArC™ Amine Reactive Compensation Bead kit (Thermo Fisher Scientific) and MACS Comp Bead Kit – anti-REA (Miltenyi Biotec) utilized. Samples were analyzed using Flowjo software v10.8 (BD Biosciences) following the gating strategy illustrated in Figure S2, the geomean fluorescence intensity was normalized to the corresponding isotype and normalized expression over 100 units was deemed positive expression of each marker as described (57 (link)).
Davies E.L., Noor M., Lim E.Y., Houldcroft C.J., Okecha G., Atkinson C., Reeves M.B., Jackson S.E, & Wills M.R. (2022). HCMV carriage in the elderly diminishes anti-viral functionality of the adaptive immune response resulting in virus replication at peripheral sites. Frontiers in Immunology, 13, 1083230.
+ Open protocol
+ Expand
3

Enumeration of Immune Cell Subsets

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The absolute number of immune cells present in whole blood samples was enumerated using Becton Dickinson Trucount tubes (BD Biosciences, Oxford, UK) following the manufacturer’s instructions. Briefly, 50µl of the EDTA treated whole blood samples was stained in the Trucount tube with a pre-mixed antibody cocktail (detailed in Table S1) allowing the identification of monocytes, B cells, CD4+ and CD8+ T cells and T cell memory subsets, NK cells and NKG2C+ NK cells. Following staining, the red blood cells were lysed and the cells fixed using FACS Lysing solution (BD Biosciences) before being stored at -80°C until acquisition (56 (link)). Samples were acquired on a 5-laser LSR Fortessa (BD Biosciences) with Fluorescence Minus One Controls and single color compensation controls (AbC Total Antibody Compensation Bead Kit – Thermo Fisher Scientific) utilized. Samples were analyzed and enumerated using Flowjo software v 10.8 (BD Biosciences) following the gating strategy and the formula illustrated in Figure S1. Results were expressed as the number of each immune cell subset per microliter of blood (cells/µl).
Davies E.L., Noor M., Lim E.Y., Houldcroft C.J., Okecha G., Atkinson C., Reeves M.B., Jackson S.E, & Wills M.R. (2022). HCMV carriage in the elderly diminishes anti-viral functionality of the adaptive immune response resulting in virus replication at peripheral sites. Frontiers in Immunology, 13, 1083230.
+ Open protocol
+ Expand
4

Multiparametric Spleen Cell Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mouse spleens were first disrupted in R10 (RPMI with 10% FBS) containing 1mg/ml collagenase D (Roche collagenase D). After 30 min, the suspension was passed through a 70 μm cell strainer and then lysed with the ACK lysis buffer system to remove erythrocytes (Lonza). The splenocytes were then washed in PBS and then stained with viability dye (Aqua or Blue Viability at 0.025 mg/ml; Thermofisher) before our staining panels were applied. Staining panels were derived from the following fluorescently conjugated anti-murine antibodies, each used at a final dilution of 1:100 in PBS: CD11c-PE Cy7 (N418, Biolegend); B220-BV605 (RA3–6B2, BD Horizon); B220-FITC (RA3–6B2, BD PharMingen); MHCII-BV510 (M5/114.15.2, Biolegend); CD3-BV785 (17A2, Biolegend); CD19-PerCP/Cy5.5 (6D5, Biolegend); IL6R-APC (D7715A7, Biolegend); CD11b-APC (M1/70, Biolegend); B220-FITC (RA3–6B2, BD PharMingen); CD138-BV421 (281–2, BD Horizon); gp130-APC (KGP130, eBioscience); CD11b-FITC (M1/70, Biolegend); Siglec-H-APC (551, Biolegend); CD93 (AA4.1, Biolegend); CD8α-BV605 (53–6.7, Biolegend); DCIR2-PE (33D1, Biolegend); Ly6C-BV650 (HK1.4, Biolegend); F4/80-Alexa 488 (BM8, Thermofisher). Samples were assayed on a 5 laser LSR Fortessa (BD Biosciences) and data was analyzed using FlowJo software version 9.3.2 (TreeStar).
Yousif A.S., Ronsard L., Shah P., Omatsu T., Sangesland M., Bracamonte Moreno T., Lam E.C., Vrbanac V.D., Balazs A.B., Reinecker H.C, & Lingwood D. (2020). The persistence of interleukin-6 is regulated by a blood buffer system derived from dendritic cells. Immunity, 54(2), 235-246.e5.
+ Open protocol
+ Expand
5

Flow Cytometric Analysis of Brain Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
The third cohort of mice was used for flow cytometry analysis. After 24 h of LPS/saline challenge, mice were euthanized by carbon dioxide (CO2) overdose, and the hippocampus and cerebellum were dissected. Dissected brain regions were gently homogenized through 70 µm cell strainers (BD Falcon) in FC buffer (ice-cold, phosphate-buffered saline (PBS) + 1% fetal calf serum). Isolated cells were blocked with 10% rat serum in ice-cold PBS for 1 h. Brain cells were stained with 0.5 μL of anti-mouse CD11b-BV421 (#101251, BioLegend, San Diego, CA, USA), CD45-BV650 (#103151, BioLegend, San Diego, CA, USA), O4-PE (#130-117-357, Miltenyi Biotec, Bergisch Gladbach, Germany), CX3CR1-A488 (#149021, BioLegend San Diego, CA, USA) for 1 h at 4 °C. Cells were washed and resuspended into 0.5 mL PBS. Samples were acquired with a 5-laser LSR Fortessa (BD Biosciences, San Jose, CA, USA) cytometer and analyzed with Kaluza v1.2 software (Beckman Coulter, Indianapolis, IN, USA).
Piirsalu M., Chithanathan K., Jayaram M., Visnapuu T., Lilleväli K., Zilmer M, & Vasar E. (2022). Lipopolysaccharide-Induced Strain-Specific Differences in Neuroinflammation and MHC-I Pathway Regulation in the Brains of Bl6 and 129Sv Mice. Cells, 11(6), 1032.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!