Moflo flow cytometer

Manufactured by Agilent Technologies

Sourced in Denmark, United States

The MoFlo flow cytometer is a high-performance instrument designed for cell analysis and sorting. It utilizes advanced laser and fluidic technologies to precisely detect and classify individual cells based on their physical and fluorescent properties. The MoFlo flow cytometer provides accurate and reliable data for a wide range of applications in the fields of biology, immunology, and cell research.

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Lab products found in correlation

Summit software, by Agilent Technologies (2 mentions) 96 well plate, by Thermo Fisher Scientific (1 mentions) Fitc brdu flow kit, by BD (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Trizol protocol, by Thermo Fisher Scientific (1 mentions) Tripsin edta, by Merck Group (1 mentions) Flowcellect annexin red kit, by Merck Group (1 mentions) Ack lysing buffer, by Lonza (1 mentions) Lysotracker, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

MoFlo cytometer (12 citations)

15 protocols using moflo flow cytometer

Flow Cytometry for Sperm Sexing

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Example 1

Now referring to FIG. 8, which shows a bivariate plot generated from the analysis of fluorochrome stained sperm cells differentiated based upon the presence of an X-chromosome or a Y-chromosome utilizing a DakoCytomation, Inc., MoFlo® flow cytometer in accordance with the invention. A conventional sheath fluid tank was retrofitted with a variable volume container in accordance with the invention containing about 5 liters of sterile sheath fluid. The sheath fluid was maintained at about 20° C. during use. An amount of gas was delivered to the sealed sheath fluid tank to exert an amount of gas pressure on the exterior surface of the variable volume container resulting in the generation of a fluid stream within the flow path of a DakoCytomation, Inc., MoFlo® flow cytometer. The flow cytometer was then otherwise operated in accordance with the standard operation procedures provided by DakoCytomation, Inc. for a period of about 8 hours to analyze and sort a mixture of sperm cells to generate a viable population of X-chromosome bearing spermatozoa and viable population of Y-chromosome bearing spermatozoa. X-chromosome bearing and Y-chromosome bearing populations enriched were established in discrete collection containers.

US10190964B2. Generating a fluid stream in a microfluidic device (2019-01-29). None [US], None [US], None [US], None [US]. Inventors: Edwin Dean Neas [US], Jerald Edward Kuiken [US], John Louis Schenk [US], Thomas Boyd Gilligan [US].

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Flow Cytometry for Sperm Sexing

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Example 1

US11175213B2. Generating a fluid stream in a microfluidic device (2021-11-16). XY, LLC [US]. Inventors: Edwin Dean Neas [US], Jerald Edward Kuiken [US], John Louis Schenk [US], Thomas Boyd Gilligan [US].

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Extracellular Vesicle Sorting and Quantification

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The MVs purified above and in suspension (20 µL) were labeled with AnnexinV-FITC (which bind phosphatidylserine (PS)) to sort total MVs, or with Annexin V-FITC and anti-CD34-PE (which bind CD34 on EPCs) to sort MVEs. The quantification and sorting were performed after 40 min incubation time, at room temperature (RT) in the dark [4 (link),6 (link)]. Fluorescent beads with diameters of 0.5, 0.9, and 3 µm were used for calibration and antibodies IgG2a-FITC and IgG1-PE were used as negative control for setting up the machine voltages. For these experiments, a flow cytometer MoFlo (Dako, Carpinteria, CA, USA) equipped with a high-speed cell sorter was used. For cell sorting by flow cytometry, the speed was about 10,000–12,000 MVs/sec.

Alexandru N., Andrei E., Safciuc F., Dragan E., Balahura A.M., Badila E, & Georgescu A. (2020). Intravenous Administration of Allogenic Cell-Derived Microvesicles of Healthy Origins Defends Against Atherosclerotic Cardiovascular Disease Development by a Direct Action on Endothelial Progenitor Cells. Cells, 9(2), 423.

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Isolation of Fetal Liver Progenitor Cells

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Single cells suspension of fetal liver was incubated in a 96 well plate (Nunc) on ice for 30 minutes with monoclonal antibodies, which are listed in Table S2. After washing, cells were incubated with streptavidin-conjugated APC-Cy7 florescent probe on ice for 20 min. The fluorescence-labeled cells were analyzed, and c-kit⁻ CD49f^+/low CD29⁺ TER119⁻ CD45⁻ cells were separated with Flowcytometer MoFlo (Dako Cytomation, Denmark; Summit version 4.0) after excluding PI⁺ dead cells.

Koike H., Ouchi R., Ueno Y., Nakata S., Obana Y., Sekine K., Zheng Y.W., Takebe T., Isono K., Koseki H, & Taniguchi H. (2014). Polycomb Group Protein Ezh2 Regulates Hepatic Progenitor Cell Proliferation and Differentiation in Murine Embryonic Liver. PLoS ONE, 9(8), e104776.

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BrdU Incorporation and DNA Fiber Analysis

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BrdU incorporation was analyzed following cell growth in six-well culture dishes and treatment with 5 mM HU or PBS control in growth medium for 1 h 37°C. After 3 PBS washes, 100 μM BrdU was added in fresh growth medium. Samples were harvested at various times following HU release, fixed and processed with the FITC BrdU Flow Kit (BD Biosciences) according to manufacturer’s directions. To correlate BrdU incorporation with cell cycle phase, cells were co-stained with propidium iodide (PI). BrdU and PI fluorescence were analyzed with a MoFlo flow cytometer and Summit software (Dako). DNA fiber analysis was performed as described [43 (link)], with three determinations and >100 fibers scored per cell type per treatment.

Ashley A.K., Shrivastav M., Nie J., Amerin C., Troksa K., Glanzer J.G., Liu S., Opiyo S.O., Dimitrova D.D., Le P., Sishc B., Bailey S.M., Oakley G.G, & Nickoloff J.A. (2014). DNA-PK Phosphorylation of RPA32 Ser4/Ser8 Regulates Replication Stress Checkpoint Activation, Fork Restart, Homologous Recombination and Mitotic Catastrophe. DNA repair, 21, 131-139.

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Isolation of Oligodendrocyte Progenitor Cells

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The following mouse monoclonal primary antibodies were used: NG2 (1:500 dilution), A2B5 (1:10) and O4 (1:3) in an adequate combination to separately harvest NG2⁺/A2B5⁻/O4⁻, NG2⁺/A2B5⁺ and NG2⁻/O4⁺ cells. Primary antibodies were directly added to the OPC proliferation medium of established secondary cultures of OPC, and incubated for 15 min at 37°C. The medium was then removed, and cells were rinse once. New OPC proliferation medium, containing the respective secondary antibodies conjugated with either Alexa Fluor 488 or Cyanine3 was added and incubated 15 min at 37°C. Cells were rinsed once, harvested by gentle pipetting, centrifuged at 100 g for 5 min and resuspended in PBS. A MoFlo flow cytometer (Dako Cytomation) was used to sort our population of interest and cells were collected in PBS. Sorted cells were finally centrifuged at 100 g for 5 min and the pellets were immediately frozen in liquid nitrogen before being store at −80°C until RNA extraction.

Gay O., Gilquin B., Assard N., Stuelsatz P., Delphin C., Lachuer J., Gidrol X, & Baudier J. (2016). Refilins are short-lived Actin-bundling proteins that regulate lamellipodium protrusion dynamics. Biology Open, 5(10), 1351-1361.

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Generation of Engineered U87-Luc/GFP Cell Line

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Human U87 glioblastoma cells that constitutively express firefly luciferase (U87-Luc) were provided by Dr. Andrew Kung (Columbia University Medical Center). In order to generate a GFP-positive U87-Luc cell line, pGIPZ lentiviral particles encoding TurboGFP (provided by Dr. Nhan Tran, TGen) were mixed with 8 µg/mL polybrene and added to subconfluent cultures of U87-Luc cells. Positively transduced cells were enriched by mass sorting the GFP-positive cells using a MoFlo flow cytometer (Dako, Carpinteria, CA). U87-Luc/GFP cells were cultured at 37°C and 5% CO₂ in DMEM (Invitrogen Corp., Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS, Invitrogen Corp.), 0.5 mg/mL G418, and 1% penicillin/streptomycin (Invitrogen Corp.). A mouse embryonic fibroblast (MEF) cell line generated from Fn14-null mice (MEF 3.5−/−) and a derivative stably transfected MEF 3.5−/− cell line expressing human Fn14 (MEF Fn14-V5) [40 (link)] were provided by Dr. Matthew Hayden (Columbia University Medical Center). Both cell lines were maintained at 37°C and 5% CO₂ in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin; the Fn14-V5 cell media also contained 10 ug/ml blasticidin.

Schneider C.S., Perez J.G., Cheng E., Zhang C., Mastorakos P., Hanes J., Winkles J.A., Woodworth G.F, & Kim A.J. (2014). Minimizing the Non-specific Binding of Nanoparticles to the Brain Enables Active Targeting of Fn14-positive Glioblastoma Cells. Biomaterials, 42, 42-51.

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Transcriptional Analysis of Chick Armc10

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EGFP-containing plasmid DNAs were electroporated and neural tubes dissected out 24 h later. Single cell suspension was obtained by 10–15 min incubation in Tripsin-EDTA (SIGMA). GFP⁺ cells were sorted by flow cytometry using a MoFlo flow cytometer (DakoCytomation). Total RNA was extracted following the Trizol protocol (Invitrogen). Reverse transcription and real-time PCR were performed according to the manufacturer's instructions (Applied Biosystems) using a PCR quantitative Real-time ABI Prism 7900HT (Applied Biosystems). Oligonucleotides specific for chick Armc10 were designed and used for amplification and normalization (Fw:5′- CAAAGCTCAAGTGCCATCAC-3′; Rv:5′-ATGCCAGCTTCTGAGCAAAT-3, Sigma). Primers specific for chick Gapdh were used for normalization. PCR amplifications were assessed from pools of electroporated neural tube chick embryos (10 embryos/pool), using two independent pools per experimental condition.

Mirra S., Ulloa F., Gutierrez-Vallejo I., Martì E, & Soriano E. (2016). Function of Armcx3 and Armc10/SVH Genes in the Regulation of Progenitor Proliferation and Neural Differentiation in the Chicken Spinal Cord. Frontiers in Cellular Neuroscience, 10, 47.

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Analysis of GO's Effect on HepG2 Cell Cycle

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Cellular effect of GO on different phases of HepG2 cell cycle was analyzed using a MoFlo flow cytometer (Dako Cytomation, Glostrup) (Nunez R, 2001). HepG2 cells (5x105 cells/well) were plated in 6-well microplates. After treatment with the double concentration of obtained GO IC50 for 24h, cells were washed twice with PBS, suspended in 300μl of PBS (pH 7.3), and finally fixed with 4ml of ice-cold 70% ethanol. To stain with propidium iodide (PI), Cells sedimentation was performed by centrifugation, the ethanol was removed and cells washed once with PBS. The cell pellets were then resuspended in 1ml of PI/Triton X-100 staining solution (0.1% Triton X-100 in PBS, 0.2 mg/ml RNase A, and10mg/ml PI) and incubated for 30 minutes at room temperature.

Loutfy S.A., Salaheldin T.A., Ramadan M.A., Farroh K.Y., Abdallah Z.F, & Youssef T. (2017). Synthesis, Characterization and Cytotoxic Evaluation of Graphene Oxide Nanosheets: In Vitro Liver Cancer Model. Asian Pacific Journal of Cancer Prevention : APJCP, 18(4), 955-961.

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Isolation and Characterization of Endothelial Progenitor Cells from Hamsters

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Peripheral blood mononuclear cells were obtained by density-gradient centrifugation using Histopaque-1077 as described by Georgescu et al. (19 (link), 30 (link)). Briefly, opaque interface containing the mononuclear cells (MNCs) was aspirate and transfer into clean tube, after centrifugation at 400 x g for 30 min, of 1 ml whole blood (collected from the retro-orbital venous sinus, ~1.5 ml per hamster) onto 3 ml Histopaque-1077. Next, the opaque interface was mixt with 10 ml isotonic PBS solution containing 2% fetal bovine serum (FBS) and centrifuged at 256 x g for 10 min. The supernatant was aspirated and discarded, and after three times repetitive washings the MNCs pellet was resuspended in 10 ml PBS.
The EPCs from C hamsters were sorted in PBS, from MNCs, at the same number, 1 x 10⁵/sample, applying the flow cytometry technique (MoFlo flow cytometer equipped with high-speed cell sorter, Dako, USA), using the specific antibodies for VEGF-R2 (KDR or Flk-1) and CD34. The CD34 and VEGF-R2 are markers for late or mature EPCs, that are found more than early EPCs (CD133⁺, CD34⁺, VEGF-R2⁺), in the peripheral circulation of adults (30 (link), 51 (link)). Importantly, the monthly blood collection from C hamsters via the retro-orbital venous sinus, to obtain EPCs, did not affect the health status of animals.

Alexandru N., Constantin A., Nemecz M., Comariţa I.K., Vîlcu A., Procopciuc A., Tanko G, & Georgescu A. (2019). Hypertension Associated With Hyperlipidemia Induced Different MicroRNA Expression Profiles in Plasma, Platelets, and Platelet-Derived Microvesicles; Effects of Endothelial Progenitor Cell Therapy. Frontiers in Medicine, 6, 280.

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