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Lsr fortessa 4 15

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The LSR Fortessa 4–15 is a flow cytometer designed for cellular analysis. It features 4 to 15 laser excitation sources, enabling the detection and measurement of multiple parameters within a single sample.

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Lab products found in correlation

Collagenase 1, by Thermo Fisher Scientific (3 mentions) Cd206, by BioLegend (2 mentions) Fv1000, by Olympus (1 mentions) Cd62l, by BD (1 mentions) Cd62l, by BioLegend (1 mentions) Accuri c6 flow cytometer, by BD (1 mentions) Lsr fortessa 4 15 hts, by BD (1 mentions)

Close spelling variants of this product

Fortessa (1232 citations) Fortessa LSR (45 citations) LSR Fortessa 4 (10 citations) LSR-Fortessa 4-15 HTS (5 citations)

5 protocols using lsr fortessa 4 15

1

Tumor-Infiltrating Immune Cell Profiling

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Tumours were harvested, treated with 1 mg/ml collagenase I (Gibco, USA) for 1 h at 37 °C. Cells were filtered through nylon mesh filters with size of 40 µm and washed with PBS. Tumour-draining lymph nodes were collected and directly ground through the cell strainers. The single-cell suspension was incubated with anti-CD16/32 (clone 93) to reduce nonspecific binding to FcRs. Cells were further stained with the following fluorochrome-conjugated antibodies: CD45 (30-F11), CD3ε (145–2C11), TCRβ (H57–597), CD4 (GK1.5), CD8α(53–6.7), Nkp46 (29A1.4), F4/80 (BM8), CD11b (M1/70), Gr-1 (RB6–8C5), CD80 (16–10A1), CD86 (GL1), CD206 (C068C2) and yellow-fluorescent reactive dye (CD45 from BD Bioscience, CD206 from Biolegend, others from eBioscience). Antibodies were used with the dilution of 1:200. Representative gating strategies for different immune cells are shown in Supplementary Fig. 54. LSR Fortessa 4–15 (BD Biosciences, USA) was used for cell acquisition and data analysis was carried out with FlowJo software (Tree Star, USA).
Ni K., Xu Z., Culbert A., Luo T., Guo N., Yang K., Pearson E., Preusser B., Wu T., Riviere P.L., Weichselbaum R.R., Spiotto M.T, & Lin W. (2022). Synergistic checkpoint-blockade and radiotherapy-radiodynamic therapy via an immunomodulatory nanoscale metal-organic framework. Nature biomedical engineering, 6(2), 144-156.
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2

Cellular Uptake and Accumulation of Cu-TBP

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The cellular uptake, intracellular accumulation and decomposition of Cu-TBP were studied systemically and compared with H4TBP. B16F10 cells were seeded on 6-well plates at 1×106/well overnight. Cu-TBP or H4TBP was added to the cells at a TBP concentration of 20 μM. After incubation of 1, 2, 4, and 8 hours, cells were collected and counted with a hemocytometer then frozen and thawed repeatly for digestion. The H4TBP was extracted with 50 μL concentrated phosphoric acid in 450 μL DMSO for UV-Vis quantification. Cells were also incubated with Cu-TBP or H4TBP for 1, 2, 4, 8 and 24 hours for detecting fluorescence of H4TBP under a confocal laser scanning microscope (CLSM, FV1000, Olympus, Japan) and further quantified by flow cytometry (LSRFortessa 4–15, BD, USA). The CLSM images were analyzed with ImageJ.
Ni K., Aung T., Li S., Fatuzzo N., Liang X, & Lin W. (2019). Nanoscale Metal-Organic Framework Mediates Radical Therapy to Enhance Cancer Immunotherapy. Chem, 5(7), 1892-1913.
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3

Characterization of Tumor-Infiltrating Immune Cells

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Tumors from syngeneic mouse models were harvested at day 12, dissected, and treated with 1 mg mL−1 collagenase I (Gibco, USA) for 30 min at 37 °C. Cells were filtered through 40 μm nylon mesh filters, centrifuged at 300 g for 10 min, and washed with PBS. The single-cell suspension was incubated with anti CD16/32 (clone 93) to reduce nonspecific binding to FcRs. Cells were further stained with the following fluorochrome-conjugated antibodies: CD45 (30-F11), CD3ϵ (145–2C11), CD4 (GK1.5), CD8α (53–6.7), PD-L1 (10F.9G2), and yellow-fluorescent reactive dye. LSR Fortessa 4–15 (BD Biosciences, USA) was used for cell acquisition and data analysis was carried out with FlowJo software (Tree Star, USA).
Li Y., Ni K., Chan C., Guo N., Luo T., Han W., Culbert A., Weichselbaum R.R, & Lin W. (2021). Dimethylaminomicheliolide Sensitizes Cancer Cells to Radiotherapy for Synergistic Combination with Immune Checkpoint Blockade. Advanced therapeutics, 5(1), 2100160.
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4

Immune cell profiling of tumor and draining lymph nodes

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Tumors and lymph nodes were harvested, treated with collagenase I (1 mg/ml) (Gibco, USA) for 1 hour at 37°C. Cells were filtered through nylon mesh filters with size of 40 μm and washed with PBS. Tumor-DLNs were collected and directly ground through the cell strainers. The single-cell suspension was incubated with anti-CD16/32 (clone 93) to reduce nonspecific binding to FcRs. Cells were further stained with the following fluorochrome-conjugated antibodies: CD45 (30-F11), CD3ε (145-2C11), CD4 (GK1.5), CD8α (53-6.7), Nkp46 (29A1.4), F4/80 (BM8), CD11b (M1/70), Gr-1 (RB6-8C5), MHC-II (AF6-120), CD80 (16-10A1), CD86 (GL1), CD206 (C068C2), CD44 (IM7), CD62L (MEL-14), H-2Kb SIINFEKL (25-D1.16), PI, and yellow fluorescent reactive dye (CD45 from BD Biosciences, CD206 and CD62L from BioLegend, and others from eBioscience). Antibodies were used with the dilution of 1:200. Representative gating strategies for different immune cells are shown in fig. S9 (F and G). LSRFortessa 4-15 (BD Biosciences, USA) was used for cell acquisition, and data analysis was carried out with FlowJo software (Tree Star, USA).
Ni K., Lan G., Guo N., Culbert A., Luo T., Wu T., Weichselbaum R.R, & Lin W. (2020). Nanoscale metal-organic frameworks for x-ray activated in situ cancer vaccination. Science Advances, 6(40), eabb5223.
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5

Flow Cytometry Data Acquisition

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Stained samples were run on the LSR Fortessa 4-15 HTS (BD Biosciences), the LSR Fortessa 4-15 (BD Biosciences), or the Accuri C6 Flow Cytometer (BD Biosciences) in the University of Chicago Cytometry and Antibody Technology Core Facility. Data were collected by FACSDiva software.
Pirozzolo I., Sepulveda M., Chen L., Wang Y., Lei Y.M., Li Z., Li R., Sattar H., Theriault B., Belkaid Y., Chong A.S, & Alegre M.L. (2022). Host-versus-commensal immune responses participate in the rejection of colonized solid organ transplants. The Journal of Clinical Investigation, 132(17), e153403.
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