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Lsrfortessa 6 laser flow cytometer

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The BD LSRFortessa 6-Laser flow cytometer is a high-performance laboratory instrument designed for multiparameter analysis of cells and other biological particles. It features six lasers that provide excitation across a wide range of fluorescent dyes, enabling the simultaneous detection and measurement of multiple cellular characteristics.

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Lab products found in correlation

Fixation permeabilization concentrate, by Thermo Fisher Scientific (2 mentions) Fc blocking ab, by Thermo Fisher Scientific (2 mentions) Anti mouse cd16 cd32, by Thermo Fisher Scientific (2 mentions) Permeabilization buffer, by Thermo Fisher Scientific (2 mentions) Fixation permeabilization diluent, by Thermo Fisher Scientific (2 mentions) Foxp3 fix perm buffer set, by BioLegend (2 mentions)

Spelling variants of this product

LSRFortessa (6355 citations) LSRFortessa flow cytometer (2456 citations) LSRFortessa 6-Laser (3 citations)

5 protocols using lsrfortessa 6 laser flow cytometer

1

Flow Cytometry Antibody Staining Protocol

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Flow cytometry was performed as previously described (27 (link)). All the conjugated antibodies were purchased from eBioscience and detailed information is given in Supplementary Table 2. Fc Blocking Ab (catalog # 14-9161-73), anti-mouse CD16/CD32 (catalog # 14-0161-82), fixation/Permeabilization concentrate (catalog# 00-5123-56), fixation/Permeabilization diluent (catalog # 00-5223-56), and permeabilization buffer (catalog# 00-8333-56) were also purchased from eBioscience. Cytometry data were acquired on a BD LSRFortessa 6-Laser flow cytometer and analyzed on Flowjo 6 software. This work was supported by the Northwestern University – Flow Cytometry Core Facility supported by Cancer Center Support Grant (NCI CA060553).
Zhai L., Bell A., Ladomersky E., Lauing K.L., Bollu L., Nguyen B., Genet M., Kim M., Chen P., Mi X., Wu J.D., Schipma M.J., Wray B., Griffiths J., Unwin R.D., Clark S.J., Acharya R., Bao R., Horbinski C., Lukas R.V., Schiltz G.E, & Wainwright D.A. (2021). Tumor Cell IDO Enhances Immune Suppression and Decreases Survival Independent of Tryptophan Metabolism in Glioblastoma. Clinical Cancer Research, 27(23), 6514-6528.
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2

Flow Cytometric Analysis of Leukocytes

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White blood cells were obtained after eliminating red blood cells by treatment three times with ammonium-chloride-potassium (ACK) lysis buffer (Invivogen). Splenocytes and LN cells were prepared as described previously. Anti-mouse CD16/CD32 was used to block FcRs and Zombie Aqua fixable viability dye was used to determine live/dead cells. For flow cytometric analysis, cells were stained using cocktails of fluorophore-conjugated anti-mouse antibodies (Table. S1). After washes, cells were suspended in cell staining buffer (eBioscience) and fixed by IC cell fixation buffer (eBioscience). Intracellular staining of Foxp3 was performed using Foxp3 Fix/Perm Buffer Set (Biolegend). Flow cytometry was performed with BD LSRFortessa 6-Laser flow cytometer (BD Biosciences) and data were analyzed with FlowJo software.
Yi S., Zhang X., Sangji H., Liu Y., Allen S.D., Xiao B., Bobbala S., Braverman C.L., Cai L., Hecker P.I., DeBerge M., Thorp E.B., Temel R.E., Stupp S.I, & Scott E.A. (2019). Surface engineered polymersomes for enhanced modulation of dendritic cells during cardiovascular immunotherapy. Advanced functional materials, 29(42), 1904399.
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3

Immune Cell Profiling in Spleen and Lymph Nodes

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After 2 months of treatment, spleen and lymph nodes (two brachial and two axillary from both sides of the mouse) were collected from all groups. Single-cell suspensions from spleen and LNs were prepared as described previously. RBC lysis buffer was used to eliminate red blood cells in spleen samples. Anti-mouse CD16/CD32 was used to block FcRs and Zombie Aqua fixable viability dye was used to determine live/dead cells. For flow cytometric analysis, cells were stained using cocktails of fluorophore-conjugated anti-mouse antibodies: BUV396 anti-CD45, FTIC anti-CD3, PerCP/Cy5.5 anti-CD4, PE anti-CD25, and Alex Fluor 647 anti-Foxp3. After washes, cells were suspended in cell staining buffer and then fixed by IC cell fixation buffer. Intracellular staining of Foxp3 was performed using Foxp3 Fix/Perm Buffer Set following the instruction (Biolegend). At least 200,000 events were recorded per tube on a BD LSRFortessa 6-Laser flow cytometer (BD Biosciences) and data were analyzed with FlowJo software.
Yi S., Karabin N.B., Zhu J., Bobbala S., Lyu H., Li S., Liu Y., Frey M., Vincent M, & Scott E.A. (2020). An Injectable Hydrogel Platform for Sustained Delivery of Anti-inflammatory Nanocarriers and Induction of Regulatory T Cells in Atherosclerosis. Frontiers in Bioengineering and Biotechnology, 8, 542.
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4

Nanoparticle Biodistribution in ApoE-/- Mice

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ApoE−/− male mice (n=4–6), were injected i.v. with 150 μl of PS, P-D2-PS, or P-D2-PEG5-PS labeled with DyLight680 (block copolymer concentration of 15 mg/ml). After 24h, mice were euthanized under CO2 anesthesia and spleen and aorta were harvested and organ NIRF imaging was performed by an IVIS Lumina with filter of 680/800 nm. The single cell suspensions were then prepared from various organs as described previously [11 (link)]. Cells were stained with anti-mouse CD16/CD32 to block FcRs and Zombie Aqua fixable viability dye prior to antibody staining. After wash, cells were then stained with multiple cocktails of fluorophore-conjugated anti-mouse antibodies (Table S1). Flow cytometry was performed with BD LSRFortessa 6-Laser flow cytometer (BD Biosciences) and data were analyzed with FlowJo software. The gating strategies were shown in Figures S13-14.
Yi S., Zhang X., Sangji H., Liu Y., Allen S.D., Xiao B., Bobbala S., Braverman C.L., Cai L., Hecker P.I., DeBerge M., Thorp E.B., Temel R.E., Stupp S.I, & Scott E.A. (2019). Surface engineered polymersomes for enhanced modulation of dendritic cells during cardiovascular immunotherapy. Advanced functional materials, 29(42), 1904399.
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5

Flow Cytometry Analysis Protocol

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Flow cytometry was performed as previously described (10 (link)). All of the conjugated antibodies were purchased from eBioscience and detailed information is given in Supplementary Table S2. Fc Blocking Ab (catalog # 14–9161–73), anti-mouse CD16/CD32 (catalog # 14–0161–82), fixation/permeabilization concentrate (catalog # 00–5123–56), fixation/permeabilization diluent (catalog # 00–5223–56), and permeabilization buffer (catalog # 00–8333–56) were also purchased from eBioscience. Cytometry data were acquired on a BD LSRFortessa 6-Laser flow cytometer and analyzed on FlowJo 6 software. This work was supported by the Northwestern University – Flow Cytometry Core Facility supported by NCI CA060553.
Zhai L., Bell A., Ladomersky E., Lauing K.L., Bollu L., Nguyen B., Genet M., Kim M., Chen P., Mi X., Wu J.D., Schipma M.J., Wray B., Griffiths J., Unwin R.D., Clark S.J., Acharya R., Bao R., Horbinski C., Lukas R.V., Schiltz G.E, & Wainwright D.A. (2021). Tumor Cell IDO Enhances Immune Suppression and Decreases Survival Independent of Tryptophan Metabolism in Glioblastoma. Clinical Cancer Research, 27(23), 6514-6528.
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