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Influx 2 cell sorter

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The Influx II cell sorter is a laboratory instrument designed for the separation and isolation of specific cell populations from complex samples. It utilizes flow cytometry technology to precisely sort cells based on their physical and fluorescent characteristics. The core function of the Influx II is to enable researchers to isolate and collect distinct cell types for further analysis and experimentation.

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Lab products found in correlation

Calcein am viability dye, by Thermo Fisher Scientific (2 mentions) Anti human cd4 microbeads, by Miltenyi Biotec (2 mentions) Anti tgfβ, by Thermo Fisher Scientific (2 mentions) Anti integrin β8 clone 37e1b5 38, by Thermo Fisher Scientific (2 mentions) Isotype control igg mopc 21, by Thermo Fisher Scientific (2 mentions) Interleukin 2 (il 2), by BioLegend (2 mentions) Active tgfβ, by Thermo Fisher Scientific (2 mentions) Anti cd3 antibody clone okt3, by BioLegend (2 mentions)

Close spelling variants of this product

BD Influx cell sorter BD Influx sorter

5 protocols using influx 2 cell sorter

1

Detailed Single-Cell Proteomics of Cell Lines

Cited 1 time
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HeLa and RAW 264.7 cells were washed by chilled PBS and sorted on the nanoPOTS chips (4 × 3 12, 1.2 mm diameter per well) using the Influx II cell sorter (BD Biosciences, San Jose, CA) as described previously (Zhu et al., 2018a (link)). To build the in-depth spectral library, 50 cells of each cell line (or equivalent peptides of ∼10 ng) were loaded onto the microPOTS chip (3 × 9, 2.2-mm diameter per well). For primary lung cells, the cells were thawed and resuspended in DMEM with 10%FBS for 1 Hr prior to be centrifuged at 800 g for 10 min. The supernatant was removed and cells were washed in DPBS. To gate out dead cells or cell debris, the cells with labeled with Calcein AM viability dye (Thermo Fisher). Similar to the FACS-sorting procedures above, we sort 50 cells into microPOTS chips for library generation and single cells into nanoPOTS chips for analysis.
Woo J., Clair G.C., Williams S.M., Feng S., Tsai C.F., Moore R.J., Chrisler W.B., Smith R.D., Kelly R.T., Paša-Tolić L., Ansong C, & Zhu Y. (2022). Three-dimensional feature matching improves coverage for single-cell proteomics based on ion mobility filtering. Cell systems, 13(5), 426-434.e4.
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2

Detailed Single-Cell Proteomics of Cell Lines

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
HeLa and RAW 264.7 cells were washed by chilled PBS and sorted on the nanoPOTS chips (4 × 3 12, 1.2 mm diameter per well) using the Influx II cell sorter (BD Biosciences, San Jose, CA) as described previously (Zhu et al., 2018a (link)). To build the in-depth spectral library, 50 cells of each cell line (or equivalent peptides of ∼10 ng) were loaded onto the microPOTS chip (3 × 9, 2.2-mm diameter per well). For primary lung cells, the cells were thawed and resuspended in DMEM with 10%FBS for 1 Hr prior to be centrifuged at 800 g for 10 min. The supernatant was removed and cells were washed in DPBS. To gate out dead cells or cell debris, the cells with labeled with Calcein AM viability dye (Thermo Fisher). Similar to the FACS-sorting procedures above, we sort 50 cells into microPOTS chips for library generation and single cells into nanoPOTS chips for analysis.
Woo J., Clair G.C., Williams S.M., Feng S., Tsai C.F., Moore R.J., Chrisler W.B., Smith R.D., Kelly R.T., Paša-Tolić L., Ansong C, & Zhu Y. (2022). Three-dimensional feature matching improves coverage for single-cell proteomics based on ion mobility filtering. Cell systems, 13(5), 426-434.e4.
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3

Single-cell isolation using Influx II

Cited 2 times
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The Influx II cell sorter (BD Biosciences, San Jose, CA) was used to isolate single cells directly into the nanowells (11 (link)). The procedures for microchip fabrication and single cell isolation were described previously (17 (link)).
Tsai C.F., Zhao R., Williams S.M., Moore R.J., Schultz K., Chrisler W.B., Pasa-Tolic L., Rodland K.D., Smith R.D., Shi T., Zhu Y, & Liu T. (2020). An Improved Boosting to Amplify Signal with Isobaric Labeling (iBASIL) Strategy for Precise Quantitative Single-cell Proteomics. Molecular & Cellular Proteomics : MCP, 19(5), 828-838.
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4

Activation of Naive CD4+ T Cells

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Human PBMCs were obtained as above, and CD4+ T cells enriched using anti-human-CD4 microbeads (Miltenyi Biotec) before sorting naive CD4+ CD45RA+ CD25−T cells by flow cytometry using an Influx II cell sorter (BD Bioscience, San Diego, CA, USA). Naive T cells were co-cultured with allogenic moDC in StemXvivo (R&D Systems) serum-free medium in roundbottom plates containing interleukin-2 (10 ng ml−1, Biolegend) and anti-CD3 antibody (clone OKT3, 0.2 µg ml−1, Biolegend). In all, 5 × 103 DCs were plated per 1 × 105 T cells in the presence of either 100 µg ml−1 anti-TGFβ (1D11), 20 µg ml−1 anti-integrin β8 (clone 37e1B5,38 100 µg ml−1 isotype control IgG (MOPC-21), or 5 ng ml−1 active TGFβ (Peprotech).
Fenton T., Kelly A., Shuttleworth E., Smedley C., Atakilit A., Powrie F., Campbell S., Nishimura S., Sheppard D., Levison S., Worthington J., Lehtinen M, & Travis M. (2016). Inflammatory cues enhance TGFβ activation by distinct subsets of human intestinal dendritic cells via integrin αvβ8. Mucosal immunology, 10(3), 624-634.
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5

Activation of Naive CD4+ T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Human PBMCs were obtained as above, and CD4+ T cells enriched using anti-human-CD4 microbeads (Miltenyi Biotec) before sorting naive CD4+ CD45RA+ CD25−T cells by flow cytometry using an Influx II cell sorter (BD Bioscience, San Diego, CA, USA). Naive T cells were co-cultured with allogenic moDC in StemXvivo (R&D Systems) serum-free medium in roundbottom plates containing interleukin-2 (10 ng ml−1, Biolegend) and anti-CD3 antibody (clone OKT3, 0.2 µg ml−1, Biolegend). In all, 5 × 103 DCs were plated per 1 × 105 T cells in the presence of either 100 µg ml−1 anti-TGFβ (1D11), 20 µg ml−1 anti-integrin β8 (clone 37e1B5,38 100 µg ml−1 isotype control IgG (MOPC-21), or 5 ng ml−1 active TGFβ (Peprotech).
Fenton T., Kelly A., Shuttleworth E., Smedley C., Atakilit A., Powrie F., Campbell S., Nishimura S., Sheppard D., Levison S., Worthington J., Lehtinen M, & Travis M. (2016). Inflammatory cues enhance TGFβ activation by distinct subsets of human intestinal dendritic cells via integrin αvβ8. Mucosal immunology, 10(3), 624-634.
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