Influx high speed sorter

Manufactured by BD

The Influx High Speed Sorter is a piece of laboratory equipment designed for the rapid and precise sorting of cells or particles. It utilizes advanced fluidic and optical technologies to enable high-throughput sorting with minimal sample loss. The core function of this product is to efficiently separate and isolate specific cell populations or particles based on their physical and fluorescent properties.

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Lab products found in correlation

Custom primers, by Merck Group (1 mentions) Mpk10025, by Thermo Fisher Scientific (1 mentions) Redtaq readymix, by Merck Group (1 mentions) Neon transfection system mpk5000, by Thermo Fisher Scientific (1 mentions) Sytox red, by Thermo Fisher Scientific (1 mentions) Ultracomp ebeads, by Thermo Fisher Scientific (1 mentions) Fixation permeabilization solution kit, by BD (1 mentions) Fcs express, by De Novo Software (1 mentions) Foxp3 transcription factor staining buffer, by Thermo Fisher Scientific (1 mentions) Mojosort human cd3 t cell isolation kit, by BioLegend (1 mentions) Fortessa, by BD (1 mentions) Lsr 2, by BD (1 mentions)

Close spelling variants of this product

Cytopeia Influx High Speed Cell Sorter Ultra-high-speed Influx cell sorter BD Influx High-Speed Cell Sorter BD Influx sorter

4 protocols using influx high speed sorter

CRISPR-Mediated Knockout of DND41 PE Cells

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DND41 PE^+/− cells were generated following a previously described protocol (59 (link)). Briefly, custom primers (Sigma-Aldrich) (Supplementary Table S3) were used to generate sgRNA DNA templates. Evaluation for potential off-targets was performed through CRISPRscan (https://www.crisprscan.org). In vitro transcription of sgRNAs from templates was performed following the protocol guidelines. 500ng of each purified sgRNA was independently incubated with 500ng of Cas9 (1074181, IDT) for 20 minutes at RT. Subsequently, one million DND41 cells were electroporated with 1μg of each complex per transfection using the Neon Transfection System (MPK5000, Thermo Fisher Scientific) and 100μL electroporating tips (MPK10025, Thermo Fisher Scientific). Electroporation conditions: pulse voltage 1350v, pulse width 10ms, pulse number 3, cell density 107/ml. Single cell clones were sorted using the Influx High Speed Sorter (BD Biosciences) and were subsequently grown in tissue culture and screened for PE loss by PCR using REDTaq ReadyMix (R2523–100RXN, Sigma Aldrich) following standard conditions and using custom primers (Supplementary Table S3).

Tottone L., Lancho O., Loh J.W., Singh A., Kimura S., Roels J., Kuchmiy A., Strubbe S., Lawlor M.A., da Silva-Diz V., Luo S., Gachet S., García-Prieto C.A., Hagelaar R., Esteller M., Meijerink J.P., Soulier J., Taghon T., Van Vlierberghe P., Mullighan C.G., Khiabanian H., Rocha P.P, & Herranz D. (2020). A Tumor Suppressor Enhancer of PTEN in T-cell Development and Leukemia. Blood Cancer Discovery, 2(1), 92-109.

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Isolation and Analysis of Lymphoid Fractions

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WKM was isolated and resuspended in PBS with 0.9% foetal bovine serum. Lymphoid and precursor fractions were sorted and analysed on an Influx high speed sorter (BD Biosciences) by utilizing their unique forward and side scatter characteristics19 (link). Sytox red (Life technologies) was used as a cell viability stain.

Wolf A., Aggio J., Campbell C., Wright F., Marquez G., Traver D, & Stachura D.L. (2017). Zebrafish Caudal Haematopoietic Embryonic Stromal Tissue (CHEST) Cells Support Haematopoiesis. Scientific Reports, 7, 44644.

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Multicolor Flow Cytometry Immunophenotyping

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For flow cytometry analysis, single-cell suspensions were stained with fluorochrome-conjugated antibodies (See Table S4 for all antibodies used in this study) in staining buffer (PBS/1% fetal bovine serum/0.1% sodium azide). Intracellular staining was performed using the Fixation/Permeabilization Solution Kit (BD Biosciences) for detection of cytokines and Foxp3/Transcription Factor Staining Buffer (Ebiosciences) for detection of transcription factors. Control samples included unstained, single fluorochrome–stained compensation beads (UltraComp eBeads, eBioscience), and fluorescence minus one (FMO) controls. Stained cells were acquired using a BD LSRII or BD Fortessa. Data were analyzed using FlowJo software (Tree Star) and FCS Express (De Novo Software). FCS Express software was used for generating t-SNE plots. For isolation of subsets by fluorescent-activated cell sorting, lymphocyte suspensions were enriched for T cells using the MojoSort Human CD3 T cell Isolation Kit (Biolegend), stained for surface markers as indicated, and sorted using the BD Influx high-speed sorter (BD Biosciences).

Kumar B.V., Ma W., Miron M., Granot T., Guyer R.S., Carpenter D.J., Senda T., Sun X., Ho S.H., Lerner H., Friedman A.L., Shen Y, & Farber D.L. (2017). Human tissue-resident memory T cells are defined by core transcriptional and functional signatures in lymphoid and mucosal sites. Cell reports, 20(12), 2921-2934.

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Isolation and Sorting of Immune Cell Subsets

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PBMC and PMN were isolated from EDTA-anti-coagulated buffy coats of healthy donors by centrifugation on double-layer histopaque-1077/1119 (Sigma). The erythrocytes in the granulocyte layer were lysed with ammonium chloride buffer. All subsequent steps were performed at 4°C. The purity of the isolated PMN typically was >90% as determined by CD16 staining. Cell sorting of mP3-positive and mP3-negative PMN was done on a BD influx high-speed sorter with use of BD Software (BD Biosciences, Franklin Lakes, New Jersey). Briefly, PMN were stained with anti-CD16-PECy7 (Clone 3G8) for 15 minutes prior to addition of mouse anti-CD177-APC (Clone MEM-166) for 30 minutes, followed by subsequent washing steps. Cells were sorted for CD177⁺CD16⁺ and CD177⁻CD16⁺ populations to obtain mP3-expressing and mP3-lacking PMN, respectively (12 (link)), the purity of the sorted PMN typically was >90%. PBMC were stained with anti-CD4-PE (Clone RPA-T4, Biolegend, San Diego, California) and anti-CD8-APC (Clone RPA-T8, Biolegend) mAbs, then CD4 and CD8 T cells were sort-purified using BD influx high-speed sorter, the purity of CD4 and CD8 cells were >95%.

Yang T.H., St. John L.S., Garber H.R., Kerros C., Ruisaard K.E., Clise-Dwyer K., Alatrash G., Ma Q, & Molldrem J.J. (2018). Membrane-Associated Proteinase 3 on Granulocytes and Acute Myeloid Leukemia Inhibits T Cell Proliferation. The Journal of Immunology Author Choice, 201(5), 1389-1399.

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