Moflo high speed sorter

Manufactured by Beckman Coulter

The MoFlo high-speed sorter is a state-of-the-art flow cytometry instrument designed for high-speed cell sorting. It is capable of efficiently sorting cells at a rate of up to 70,000 events per second.

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Lab products found in correlation

Xyclone, by Hamilton Thorne (1 mentions) Moflo astrio, by Beckman Coulter (1 mentions) Fluo 4 nw, by Thermo Fisher Scientific (1 mentions) Dynabeads untouched mouse cd4 cells kit, by Thermo Fisher Scientific (1 mentions) Lsrii flow cytometer, by BD (1 mentions)

Close spelling variants of this product

MoFlo XDP high-speed flow cytometry sorter MoFlo Astrios EQ high speed cell sorter MoFlo high-speed cell sorter Legacy MoFlo MLS High Speed Cell Sorter MoFlo XDP high-speed sorter MoFlo MLS high-speed cell sorter MoFlo XDP High-Speed Cell Sorter MoFlo Astrios High Speed Sorter MoFlo sorter

3 protocols using moflo high speed sorter

Embryo Harvesting and ESC Injection

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Embryos were harvested from F1 (C57BL/6×CBA) or C57BL/6×F1 crosses. Females were selected by morphological identification of oestrus (Champlin et al., 1973 (link)). Detection of a copulation plug on the following day confirmed mating. Embryo staging was based on the assumption that mating occurred at midnight, so that at 12 noon the next day embryos are assigned E (embryonic day) 0.5 or 12 hpc. E2.5 (60 hpc) embryos were flushed from oviducts in M2 (Sigma) and cultured in BlastAssist (Origio) under embryo-tested mineral oil (Sigma) at 37°C and 7% CO₂ in air. ESCs (3-8) were injected via a laser-generated perforation in the zona pellucida using XYClone (Hamilton Thorne Biosciences). For experiments comparing naive pluripotent versus differentiating donor cells, ESCs were sorted for Rex1-GFPd2^high (top 5% of population) or Rex1-GFPd2^low (bottom 5% of population) expression using a Beckman Coulter MoFlo high-speed sorter immediately before injection.

Alexandrova S., Kalkan T., Humphreys P., Riddell A., Scognamiglio R., Trumpp A, & Nichols J. (2016). Selection and dynamics of embryonic stem cell integration into early mouse embryos. Development (Cambridge, England), 143(1), 24-34.

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Purification of Hematopoietic Stem and Progenitor Cells

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Prior to sorting, stained bone marrow cells were suspended in flow cytometry buffer containing 10% FCS, filtered using Partec CellTrics sterile filters (Sysmex-Partec, Görlitz, Germany) and stored on ice. All of the HSPC populations were sorted twice to ensure high purity. Cells were sorted into LightCycler 480 384-well plates (Roche, Basel, Switzerland) for qRT-PCR analysis or into culture medium for in vitro manipulation. Sorting was carried out using either a MoFlo High Speed Sorter (Beckman Coulter) controlled by Summit v4.3 software or a MoFlo Astrios (Beckman Coulter) controlled by Summit v6.2.3 software.

Mooney C.J., Cunningham A., Tsapogas P., Toellner K.M, & Brown G. (2017). Selective Expression of Flt3 within the Mouse Hematopoietic Stem Cell Compartment. International Journal of Molecular Sciences, 18(5), 1037.

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Murine T Cell Isolation and Signaling

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After red blood cell lysis, murine LN and spleen T cells were enriched to >90% purity using Dynabeads Untouched Mouse CD4 Cells Kit (Invitrogen). For some experiments, naïve cells (CD44^lo CD62L^hi) were sorted using a MoFlo high-speed sorter (Beckman Coulter). Cells were analyzed on a LSR II flow cytometer (BD) and analyzed with FlowJo software (Tree Star).
For TCR proximal signaling experiments, purified T cells were stimulated with anti-CD3 mAb followed by crosslinking with goat anti-hamster IgG for the indicated times at 37°C with gentle shaking. For stimulations >1 h, purified T cells were stimulated with plate-coated anti-CD3 and anti-CD28, PMA plus ionomycin or concanavalin A as indicated. Calcium flux was measured using Fluo-4 NW (Invitrogen) according to the manufacturer’s instructions with modifications. Briefly, 4 × 10⁶ purified CD4 T cells were loaded with Fluo-4 NW dye mix for 30 min at 37°C followed by 30 min at 25°C. After labeling, cells were recorded for background fluorescence by flow cytometry before addition of anti-CD3 at 30 s and goat anti-hamster IgG at 120 s. Detailed protocols are described in Supplemental Experimental Procedures.

Whang M.I., Tavares R.M., Benjamin D.I., Kattah M.G., Advincula R., Nomura D.K., Debnath J., Malynn B.A, & Ma A. (2017). The Ubiquitin Binding Protein TAX1BP1 Mediates Autophagasome Induction and the Metabolic Transition of Activated T Cells. Immunity, 46(3), 405-420.

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