flow cytometry instrument  (Millipore)


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    Name:
    Flow Cytometry Instruments
    Description:
    Cell by cell analysis tailored to your research needs available in three flow cytometry platforms Muse Cell AnalyzerThe Muse system which uses microcapillary fluidics and pre optimized reagents requires little setup or expertise Applications include cell viability apoptosis cell cycle progression DNA damage response cell cycle effects and signaling pathway activation Guava Benchtop Microcapillary Flow CytometersCompared to traditional sheath fluid based instruments the easy to use Guava flow cytometers consume less sample and produce less liquid waste Powerful violet blue green and red lasers offer compatibility with a wide range of fluorochromes Amnis Imaging Flow CytometersAmnis imaging flow cytometers combine microscopy and flow cytometry to address complex biological questions unresolved by traditional flow cytometers The sensitive Amnis instruments enable the detection of rare events and reveal the spatial location of biomolecules and cellular components
    Catalog Number:
    flowcyt
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    Structured Review

    Millipore flow cytometry instrument
    Flow Cytometry Instruments
    Cell by cell analysis tailored to your research needs available in three flow cytometry platforms Muse Cell AnalyzerThe Muse system which uses microcapillary fluidics and pre optimized reagents requires little setup or expertise Applications include cell viability apoptosis cell cycle progression DNA damage response cell cycle effects and signaling pathway activation Guava Benchtop Microcapillary Flow CytometersCompared to traditional sheath fluid based instruments the easy to use Guava flow cytometers consume less sample and produce less liquid waste Powerful violet blue green and red lasers offer compatibility with a wide range of fluorochromes Amnis Imaging Flow CytometersAmnis imaging flow cytometers combine microscopy and flow cytometry to address complex biological questions unresolved by traditional flow cytometers The sensitive Amnis instruments enable the detection of rare events and reveal the spatial location of biomolecules and cellular components
    https://www.bioz.com/result/flow cytometry instrument/product/Millipore
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    flow cytometry instrument - by Bioz Stars, 2020-05
    99/100 stars

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    Related Articles

    Flow Cytometry:

    Article Title: Cdt1 variants reveal unanticipated aspects of interactions with cyclin/CDK and MCM important for normal genome replication
    Article Snippet: .. Flow cytometry analysis for DNA rereplication U2OS cell lines harboring stably integrated individual Cdt1 alleles were cultured in complete medium plus doxycycline for either 48 or 72 h. Cell were pulse-labeled with 10 µM EdU (Sigma) for 30 min before being harvested by trypsinization. ..

    Article Title: TNF blockade induces a dysregulated type I interferon response without autoimmunity in paradoxical psoriasis
    Article Snippet: .. For flow cytometry analysis, mouse skin was digested with Dispase (Sigma-Aldrich) and collagenase (Invitrogen), and stained with anti-B220 FITC (BD Pharmingen, RA3-6B2, 1/400), anti-CD45 PerCp-Cy5.5 (BD Pharmingen, 30-F11, 1/400), anti-CD11c PE (eBioscience, N418, 1/800), and anti-PDCA1 APC (Biolegend, 927, 1/400), anti-CD80 PE (BD Pharmingen, 16-10A1, 1/800), or anti-CD86 PE (BD Pharmingen, GL1, 1/800). .. For flow cytometry analyses of human pDCs, antibodies used include anti-CD123 APC (Biolegend, 6H6, 1/400), anti-BDCA2 PE (Miltenyi, AC144, 1/400), anti-BDCA4 APC (Miltenyi, REA-380, 1/400), anti-CD80 PE (BD Pharmingen, 16-10A1, 1/800), anti-CD83 FITC (eBioscience, HB15e, 1/400), and anti-HLA-DR (BD Pharmingen, G46-6, 1/400).

    Article Title: Integrase Defective Lentiviral Vector as a Vaccine Platform for Delivering Influenza Antigens
    Article Snippet: .. Flow Cytometry and Confocal Laser Scanning Microscopy (CLSM) 293 T Lenti-X cells were plated in six-well microplates for flow cytometry analysis or seeded in 24-well cluster plates onto 12-mm cover glasses previously treated with l -polylysine (SIGMA) for CLSM. .. Cells were then transfected by calcium phosphate with pCMVHACal09 using the Profection Mammalian Transfection System (Promega).

    Article Title: Th17 can regulate silica-induced lung inflammation through an IL-1β-dependent mechanism
    Article Snippet: .. Flow cytometry analysis The cells from spleen and HLNs were stimulated with phorbol-12-myristate-13-acetate (50 ng/ml) and ionomycin (1 mg/ml; Sigma-Aldrich, Palo Alto, CA, USA) for 4 hrs, and monensin (BD GolgiStop Protein Transport Inhibitor) was added, as described previously [ ]. .. Fc receptors (FcRs) were blocked with purified rat antimouse CD16/CD32 (BD Pharmingen, San Jose, CA, USA).

    Article Title: The Study of a Novel Sorafenib Derivative HLC-080 as an Antitumor Agent
    Article Snippet: .. Flow cytometry analysis Cells were incubated with either Sorafenib, HLC-080 or DMSO (control) for 96 h. Then, the cells were washed twice with ice-cold phosphate buffered saline (PBS), harvested, fixed with ice-cold PBS in 70% ethanol, and stored at −20°C for 2 h. After fixation, the cells were incubated with RNase A (Sigma), stained with propidium iodide (Sigma) for 30 min in the dark. .. DNA content was analyzed by flow cytometric analysis.

    Article Title: Downregulated miR-45 Inhibits the G1-S Phase Transition by Targeting Bmi-1 in Breast Cancer
    Article Snippet: .. Flow Cytometry Analysis According to previous report, 20,000 harvested cells were washed, fixed, pelleted, re-suspended, incubated with bovine pancreatic RNAase (Sigma, Saint Louis, MO), and stained with propidium iodide (Sigma-Aldrich) before analyzed on a flow cytometer (FACSCalibur; BD Biosciences). .. Luciferase Assays According to the manufacturer's recommendation of the Lipofectamine 2000 reagent (Invitrogen Co, Carlsbad, CA), 100 ng of p3x IRSMLP -luciferase plasmid, or pGL3-Bmi-1–3′ UTR (wt/mut), or the control-luciferase plasmid, plus 1ng of pRL-TK renilla plasmid (Promega, Madison, WI) was transfected into the indicated cells.

    Article Title: Growth Factor Screening in Dystrophic Muscles Reveals PDGFB/PDGFRB-Mediated Migration of Interstitial Stem Cells
    Article Snippet: .. Flow Cytometry Analysis of Murine Cells The same number of cells was seeded into flat-bottom T75 cell culture flasks (Sigma-Aldrich #CLS3290) coated with 0.1 percent Collagen (Sigma-Aldrich #C9791), cultured to confluence in high glucose DMEM medium (Gibco #61965026) supplemented with 20 percent FBS (Gibco #10082147), 10 percent HS (Gibco #26050088) and 1 percent penicillin–streptomycin (Gibco #15140122). .. The single-cell suspension was diluted to 1 × 106 cells/mL with FACS buffer (2 percent foetal bovine serum (FBS, Gibco #10082147), 10mM HEPES and 10mM NaN3 at a pH of 7.2) and supplemented for 30 min at 4 °C protected from light with the following antibodies: anti-mouse CCR1-APC (R & D systems #FAB5986A), anti-mouse CCR5-PE (Invitrogen #12-1951-82), anti-mouse PDGFRA-APC (Invitrogen #17-1401-81) and anti-mouse PDGFRB-APC (Invitrogen #17-1402-82), anti-mouse CD34-eFluor450 (Invitrogen #48-0341-82), anti-mouse CD44 (Invitrogen #17-0441-82).

    Article Title: Therapeutic effects of thalidomide in myeloma are associated with the expression of fibroblast growth factor receptor 3
    Article Snippet: .. Apoptosis and flow cytometry analysis 1 × 106 MM cells, cultured 48 hours after treatment with Thal were harvested, washed with PBS, fixed with 70% ethanol, and pretreated with 10 mg/mL of RNase (Sigma). .. Cells were stained with propidium iodide (PI; 10 mg/mL, Sigma), and analyzed by flow cytometry.

    Stable Transfection:

    Article Title: Cdt1 variants reveal unanticipated aspects of interactions with cyclin/CDK and MCM important for normal genome replication
    Article Snippet: .. Flow cytometry analysis for DNA rereplication U2OS cell lines harboring stably integrated individual Cdt1 alleles were cultured in complete medium plus doxycycline for either 48 or 72 h. Cell were pulse-labeled with 10 µM EdU (Sigma) for 30 min before being harvested by trypsinization. ..

    Cytometry:

    Article Title: Cdt1 variants reveal unanticipated aspects of interactions with cyclin/CDK and MCM important for normal genome replication
    Article Snippet: .. Flow cytometry analysis for DNA rereplication U2OS cell lines harboring stably integrated individual Cdt1 alleles were cultured in complete medium plus doxycycline for either 48 or 72 h. Cell were pulse-labeled with 10 µM EdU (Sigma) for 30 min before being harvested by trypsinization. ..

    Article Title: TNF blockade induces a dysregulated type I interferon response without autoimmunity in paradoxical psoriasis
    Article Snippet: .. For flow cytometry analysis, mouse skin was digested with Dispase (Sigma-Aldrich) and collagenase (Invitrogen), and stained with anti-B220 FITC (BD Pharmingen, RA3-6B2, 1/400), anti-CD45 PerCp-Cy5.5 (BD Pharmingen, 30-F11, 1/400), anti-CD11c PE (eBioscience, N418, 1/800), and anti-PDCA1 APC (Biolegend, 927, 1/400), anti-CD80 PE (BD Pharmingen, 16-10A1, 1/800), or anti-CD86 PE (BD Pharmingen, GL1, 1/800). .. For flow cytometry analyses of human pDCs, antibodies used include anti-CD123 APC (Biolegend, 6H6, 1/400), anti-BDCA2 PE (Miltenyi, AC144, 1/400), anti-BDCA4 APC (Miltenyi, REA-380, 1/400), anti-CD80 PE (BD Pharmingen, 16-10A1, 1/800), anti-CD83 FITC (eBioscience, HB15e, 1/400), and anti-HLA-DR (BD Pharmingen, G46-6, 1/400).

    Article Title: Integrase Defective Lentiviral Vector as a Vaccine Platform for Delivering Influenza Antigens
    Article Snippet: .. Flow Cytometry and Confocal Laser Scanning Microscopy (CLSM) 293 T Lenti-X cells were plated in six-well microplates for flow cytometry analysis or seeded in 24-well cluster plates onto 12-mm cover glasses previously treated with l -polylysine (SIGMA) for CLSM. .. Cells were then transfected by calcium phosphate with pCMVHACal09 using the Profection Mammalian Transfection System (Promega).

    Article Title: Th17 can regulate silica-induced lung inflammation through an IL-1β-dependent mechanism
    Article Snippet: .. Flow cytometry analysis The cells from spleen and HLNs were stimulated with phorbol-12-myristate-13-acetate (50 ng/ml) and ionomycin (1 mg/ml; Sigma-Aldrich, Palo Alto, CA, USA) for 4 hrs, and monensin (BD GolgiStop Protein Transport Inhibitor) was added, as described previously [ ]. .. Fc receptors (FcRs) were blocked with purified rat antimouse CD16/CD32 (BD Pharmingen, San Jose, CA, USA).

    Article Title: The Study of a Novel Sorafenib Derivative HLC-080 as an Antitumor Agent
    Article Snippet: .. Flow cytometry analysis Cells were incubated with either Sorafenib, HLC-080 or DMSO (control) for 96 h. Then, the cells were washed twice with ice-cold phosphate buffered saline (PBS), harvested, fixed with ice-cold PBS in 70% ethanol, and stored at −20°C for 2 h. After fixation, the cells were incubated with RNase A (Sigma), stained with propidium iodide (Sigma) for 30 min in the dark. .. DNA content was analyzed by flow cytometric analysis.

    Article Title: Downregulated miR-45 Inhibits the G1-S Phase Transition by Targeting Bmi-1 in Breast Cancer
    Article Snippet: .. Flow Cytometry Analysis According to previous report, 20,000 harvested cells were washed, fixed, pelleted, re-suspended, incubated with bovine pancreatic RNAase (Sigma, Saint Louis, MO), and stained with propidium iodide (Sigma-Aldrich) before analyzed on a flow cytometer (FACSCalibur; BD Biosciences). .. Luciferase Assays According to the manufacturer's recommendation of the Lipofectamine 2000 reagent (Invitrogen Co, Carlsbad, CA), 100 ng of p3x IRSMLP -luciferase plasmid, or pGL3-Bmi-1–3′ UTR (wt/mut), or the control-luciferase plasmid, plus 1ng of pRL-TK renilla plasmid (Promega, Madison, WI) was transfected into the indicated cells.

    Article Title: Growth Factor Screening in Dystrophic Muscles Reveals PDGFB/PDGFRB-Mediated Migration of Interstitial Stem Cells
    Article Snippet: .. Flow Cytometry Analysis of Murine Cells The same number of cells was seeded into flat-bottom T75 cell culture flasks (Sigma-Aldrich #CLS3290) coated with 0.1 percent Collagen (Sigma-Aldrich #C9791), cultured to confluence in high glucose DMEM medium (Gibco #61965026) supplemented with 20 percent FBS (Gibco #10082147), 10 percent HS (Gibco #26050088) and 1 percent penicillin–streptomycin (Gibco #15140122). .. The single-cell suspension was diluted to 1 × 106 cells/mL with FACS buffer (2 percent foetal bovine serum (FBS, Gibco #10082147), 10mM HEPES and 10mM NaN3 at a pH of 7.2) and supplemented for 30 min at 4 °C protected from light with the following antibodies: anti-mouse CCR1-APC (R & D systems #FAB5986A), anti-mouse CCR5-PE (Invitrogen #12-1951-82), anti-mouse PDGFRA-APC (Invitrogen #17-1401-81) and anti-mouse PDGFRB-APC (Invitrogen #17-1402-82), anti-mouse CD34-eFluor450 (Invitrogen #48-0341-82), anti-mouse CD44 (Invitrogen #17-0441-82).

    Article Title: Therapeutic effects of thalidomide in myeloma are associated with the expression of fibroblast growth factor receptor 3
    Article Snippet: .. Apoptosis and flow cytometry analysis 1 × 106 MM cells, cultured 48 hours after treatment with Thal were harvested, washed with PBS, fixed with 70% ethanol, and pretreated with 10 mg/mL of RNase (Sigma). .. Cells were stained with propidium iodide (PI; 10 mg/mL, Sigma), and analyzed by flow cytometry.

    Cell Culture:

    Article Title: Cdt1 variants reveal unanticipated aspects of interactions with cyclin/CDK and MCM important for normal genome replication
    Article Snippet: .. Flow cytometry analysis for DNA rereplication U2OS cell lines harboring stably integrated individual Cdt1 alleles were cultured in complete medium plus doxycycline for either 48 or 72 h. Cell were pulse-labeled with 10 µM EdU (Sigma) for 30 min before being harvested by trypsinization. ..

    Article Title: Growth Factor Screening in Dystrophic Muscles Reveals PDGFB/PDGFRB-Mediated Migration of Interstitial Stem Cells
    Article Snippet: .. Flow Cytometry Analysis of Murine Cells The same number of cells was seeded into flat-bottom T75 cell culture flasks (Sigma-Aldrich #CLS3290) coated with 0.1 percent Collagen (Sigma-Aldrich #C9791), cultured to confluence in high glucose DMEM medium (Gibco #61965026) supplemented with 20 percent FBS (Gibco #10082147), 10 percent HS (Gibco #26050088) and 1 percent penicillin–streptomycin (Gibco #15140122). .. The single-cell suspension was diluted to 1 × 106 cells/mL with FACS buffer (2 percent foetal bovine serum (FBS, Gibco #10082147), 10mM HEPES and 10mM NaN3 at a pH of 7.2) and supplemented for 30 min at 4 °C protected from light with the following antibodies: anti-mouse CCR1-APC (R & D systems #FAB5986A), anti-mouse CCR5-PE (Invitrogen #12-1951-82), anti-mouse PDGFRA-APC (Invitrogen #17-1401-81) and anti-mouse PDGFRB-APC (Invitrogen #17-1402-82), anti-mouse CD34-eFluor450 (Invitrogen #48-0341-82), anti-mouse CD44 (Invitrogen #17-0441-82).

    Article Title: Therapeutic effects of thalidomide in myeloma are associated with the expression of fibroblast growth factor receptor 3
    Article Snippet: .. Apoptosis and flow cytometry analysis 1 × 106 MM cells, cultured 48 hours after treatment with Thal were harvested, washed with PBS, fixed with 70% ethanol, and pretreated with 10 mg/mL of RNase (Sigma). .. Cells were stained with propidium iodide (PI; 10 mg/mL, Sigma), and analyzed by flow cytometry.

    Incubation:

    Article Title: The Study of a Novel Sorafenib Derivative HLC-080 as an Antitumor Agent
    Article Snippet: .. Flow cytometry analysis Cells were incubated with either Sorafenib, HLC-080 or DMSO (control) for 96 h. Then, the cells were washed twice with ice-cold phosphate buffered saline (PBS), harvested, fixed with ice-cold PBS in 70% ethanol, and stored at −20°C for 2 h. After fixation, the cells were incubated with RNase A (Sigma), stained with propidium iodide (Sigma) for 30 min in the dark. .. DNA content was analyzed by flow cytometric analysis.

    Article Title: Downregulated miR-45 Inhibits the G1-S Phase Transition by Targeting Bmi-1 in Breast Cancer
    Article Snippet: .. Flow Cytometry Analysis According to previous report, 20,000 harvested cells were washed, fixed, pelleted, re-suspended, incubated with bovine pancreatic RNAase (Sigma, Saint Louis, MO), and stained with propidium iodide (Sigma-Aldrich) before analyzed on a flow cytometer (FACSCalibur; BD Biosciences). .. Luciferase Assays According to the manufacturer's recommendation of the Lipofectamine 2000 reagent (Invitrogen Co, Carlsbad, CA), 100 ng of p3x IRSMLP -luciferase plasmid, or pGL3-Bmi-1–3′ UTR (wt/mut), or the control-luciferase plasmid, plus 1ng of pRL-TK renilla plasmid (Promega, Madison, WI) was transfected into the indicated cells.

    Confocal Laser Scanning Microscopy:

    Article Title: Integrase Defective Lentiviral Vector as a Vaccine Platform for Delivering Influenza Antigens
    Article Snippet: .. Flow Cytometry and Confocal Laser Scanning Microscopy (CLSM) 293 T Lenti-X cells were plated in six-well microplates for flow cytometry analysis or seeded in 24-well cluster plates onto 12-mm cover glasses previously treated with l -polylysine (SIGMA) for CLSM. .. Cells were then transfected by calcium phosphate with pCMVHACal09 using the Profection Mammalian Transfection System (Promega).

    Staining:

    Article Title: TNF blockade induces a dysregulated type I interferon response without autoimmunity in paradoxical psoriasis
    Article Snippet: .. For flow cytometry analysis, mouse skin was digested with Dispase (Sigma-Aldrich) and collagenase (Invitrogen), and stained with anti-B220 FITC (BD Pharmingen, RA3-6B2, 1/400), anti-CD45 PerCp-Cy5.5 (BD Pharmingen, 30-F11, 1/400), anti-CD11c PE (eBioscience, N418, 1/800), and anti-PDCA1 APC (Biolegend, 927, 1/400), anti-CD80 PE (BD Pharmingen, 16-10A1, 1/800), or anti-CD86 PE (BD Pharmingen, GL1, 1/800). .. For flow cytometry analyses of human pDCs, antibodies used include anti-CD123 APC (Biolegend, 6H6, 1/400), anti-BDCA2 PE (Miltenyi, AC144, 1/400), anti-BDCA4 APC (Miltenyi, REA-380, 1/400), anti-CD80 PE (BD Pharmingen, 16-10A1, 1/800), anti-CD83 FITC (eBioscience, HB15e, 1/400), and anti-HLA-DR (BD Pharmingen, G46-6, 1/400).

    Article Title: The Study of a Novel Sorafenib Derivative HLC-080 as an Antitumor Agent
    Article Snippet: .. Flow cytometry analysis Cells were incubated with either Sorafenib, HLC-080 or DMSO (control) for 96 h. Then, the cells were washed twice with ice-cold phosphate buffered saline (PBS), harvested, fixed with ice-cold PBS in 70% ethanol, and stored at −20°C for 2 h. After fixation, the cells were incubated with RNase A (Sigma), stained with propidium iodide (Sigma) for 30 min in the dark. .. DNA content was analyzed by flow cytometric analysis.

    Article Title: Downregulated miR-45 Inhibits the G1-S Phase Transition by Targeting Bmi-1 in Breast Cancer
    Article Snippet: .. Flow Cytometry Analysis According to previous report, 20,000 harvested cells were washed, fixed, pelleted, re-suspended, incubated with bovine pancreatic RNAase (Sigma, Saint Louis, MO), and stained with propidium iodide (Sigma-Aldrich) before analyzed on a flow cytometer (FACSCalibur; BD Biosciences). .. Luciferase Assays According to the manufacturer's recommendation of the Lipofectamine 2000 reagent (Invitrogen Co, Carlsbad, CA), 100 ng of p3x IRSMLP -luciferase plasmid, or pGL3-Bmi-1–3′ UTR (wt/mut), or the control-luciferase plasmid, plus 1ng of pRL-TK renilla plasmid (Promega, Madison, WI) was transfected into the indicated cells.

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  • 99
    Millipore flow cytometry analysis
    Characterization of the Sf Basic cell line and comparison to the parental Sf 21 cell line. ( A ) Cell diameter (Ø) distribution histogram with a representative bright field image. ( B ) Fluorescence microscopy images of propidium iodide stained cells. Mean fluorescence intensity (MFI) of the G1-phase cell population from flow <t>cytometry</t> cell cycle analysis of the same sample is indicated below the corresponding image. ( C ) Cell line population doubling time (PDT) as a function of growth medium pH. The growth medium was Sf-900II-BES-MISS, adjusted to various pH levels by titration with 1 N NaOH.
    Flow Cytometry Analysis, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 395 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/flow cytometry analysis/product/Millipore
    Average 99 stars, based on 395 article reviews
    Price from $9.99 to $1999.99
    flow cytometry analysis - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    88
    Millipore i selex device
    Optimization of the operating flow rates within the <t>I-SELEX</t> device to achieve stringent separation of a model non-interacting aptamer from t <t>RBCs</t> while maximizing t RBC recovery. ( a ) Average fluorescence intensity line scans showing the normalized distribution of unbound FITC-labeled CRP aptamers (200 nM) across the channel width at increasing flow rates. Approximate positions of the product and waste outlets are indicated. Corresponding fluorescence images illustrating flow positions of unbound aptamers are also shown as an inset (yellow dashed lines indicate the approximate position of the microchannel walls). ( b ) Average composite images indicate unbound aptamers move to the outer wall and are diverted into the waste outer outlet. ( c ) Average composite images indicate efficient t RBC focusing to the inner microchannel wall and diversion into the product inner outlet. In both ( b ) and ( c ), the sample input and sheath buffer flow rates are 150 μL min −1 and 1500 μL min −1 , respectively, and the yellow dashed lines indicate approximate positions of the microchannel walls and bifurcation. ( d ) Recovery of t RBCs at the product outlet as a percentage of the cells loaded into the device when operated at different sample input flow rates. In all cases, the sheath buffer flow rate is 10-fold higher than the sample input flow rate.
    I Selex Device, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/i selex device/product/Millipore
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    i selex device - by Bioz Stars, 2020-05
    88/100 stars
      Buy from Supplier

    Image Search Results


    Characterization of the Sf Basic cell line and comparison to the parental Sf 21 cell line. ( A ) Cell diameter (Ø) distribution histogram with a representative bright field image. ( B ) Fluorescence microscopy images of propidium iodide stained cells. Mean fluorescence intensity (MFI) of the G1-phase cell population from flow cytometry cell cycle analysis of the same sample is indicated below the corresponding image. ( C ) Cell line population doubling time (PDT) as a function of growth medium pH. The growth medium was Sf-900II-BES-MISS, adjusted to various pH levels by titration with 1 N NaOH.

    Journal: PLoS ONE

    Article Title: Enhanced Production of Chikungunya Virus-Like Particles Using a High-pH Adapted Spodoptera frugiperda Insect Cell Line

    doi: 10.1371/journal.pone.0094401

    Figure Lengend Snippet: Characterization of the Sf Basic cell line and comparison to the parental Sf 21 cell line. ( A ) Cell diameter (Ø) distribution histogram with a representative bright field image. ( B ) Fluorescence microscopy images of propidium iodide stained cells. Mean fluorescence intensity (MFI) of the G1-phase cell population from flow cytometry cell cycle analysis of the same sample is indicated below the corresponding image. ( C ) Cell line population doubling time (PDT) as a function of growth medium pH. The growth medium was Sf-900II-BES-MISS, adjusted to various pH levels by titration with 1 N NaOH.

    Article Snippet: A Guava Cell Cycle Kit (Millipore) was used to stain the cells for flow cytometry analysis using a Guava EasyCyte8HT capillary flow cytometer (Millipore) and the manufacturer-supplied cell cycle procedure.

    Techniques: Fluorescence, Microscopy, Staining, Flow Cytometry, Cytometry, Cell Cycle Assay, Titration

    Orientation of NHis–Cx43 in the liposomal membrane. a) Schematic illustration of the orientation of Cx43 in living systems. The N‐terminus, CL, and C‐terminus of Cx43 are positioned at the cytoplasm. ELs protrude into the extracellular space. b) Schematic illustration of the detectable proteolytic fragments (top) and western blot analysis of proteolytic fragments of Cx43 and NHis–Cx43, using Cx43‐specific antibodies after trypsin treatment (bottom). The arrows indicate the full‐length Cx43 and NHis–Cx43 (FL) and a proteolytic fragment (a). Results are representative of three independent experiments. c) Schematic illustration of Cx43–EL1FLAG and NHis–Cx43–EL1FLAG fusion proteins. The black arrow indicates the His tag and the red arrow indicates the FLAG tag. d) Schematic illustration of flow cytometry following immunoprecipitation. e) Flow cytometry analysis of Cx43–EL1FLAG‐ or NHis–Cx43–EL1FLAG‐integrated proteoliposomes (green area), using a FLAG‐tag‐specific antibody. Solid lines indicate loading of FLAG‐tag‐specific antibody‐modified beads. Red and blue lines indicate loading of beads reacted with Cx43‐integrated proteoliposomes and NHis–Cx43‐integrated proteoliposomes, respectively.

    Journal: Advanced Science

    Article Title: Proteoliposome Engineering with Cell‐Free Membrane Protein Synthesis: Control of Membrane Protein Sorting into Liposomes by Chaperoning Systems

    doi: 10.1002/advs.201800524

    Figure Lengend Snippet: Orientation of NHis–Cx43 in the liposomal membrane. a) Schematic illustration of the orientation of Cx43 in living systems. The N‐terminus, CL, and C‐terminus of Cx43 are positioned at the cytoplasm. ELs protrude into the extracellular space. b) Schematic illustration of the detectable proteolytic fragments (top) and western blot analysis of proteolytic fragments of Cx43 and NHis–Cx43, using Cx43‐specific antibodies after trypsin treatment (bottom). The arrows indicate the full‐length Cx43 and NHis–Cx43 (FL) and a proteolytic fragment (a). Results are representative of three independent experiments. c) Schematic illustration of Cx43–EL1FLAG and NHis–Cx43–EL1FLAG fusion proteins. The black arrow indicates the His tag and the red arrow indicates the FLAG tag. d) Schematic illustration of flow cytometry following immunoprecipitation. e) Flow cytometry analysis of Cx43–EL1FLAG‐ or NHis–Cx43–EL1FLAG‐integrated proteoliposomes (green area), using a FLAG‐tag‐specific antibody. Solid lines indicate loading of FLAG‐tag‐specific antibody‐modified beads. Red and blue lines indicate loading of beads reacted with Cx43‐integrated proteoliposomes and NHis–Cx43‐integrated proteoliposomes, respectively.

    Article Snippet: Flow Cytometry Analysis of the Topology of Integrated Cx43 Derivatives : Preparation of proteoliposomes integrated with Cx43 derivatives was performed with or without molecular chaperones containing DnaK, DnaJ, and GrpE (DnaK mix; GeneFrontier) as described under subsection “Preparation and Purification of Membrane Protein‐Integrated Proteoliposomes.” PureProteome Protein G magnetic beads (Millipore) (2 × 105 particles per tube) were incubated with 20 µg mL−1 mouse anti‐FLAG M2 IgG (Sigma‐Aldrich, St. Louis, MO, USA) with rotation for 1 h at room temperature.

    Techniques: Western Blot, FLAG-tag, Flow Cytometry, Cytometry, Immunoprecipitation, Modification

    Graphical representation demonstrating the flow cytometry percentage for CD44/CD24 cancer stem cell phenotype in cells. Flow citometry for cells grown in mololayer for control group of (A) CMTU-229 cell line and (B) MCF-7 cell line. Flow citometry for mammospheres for both cell lines (C) CMTU-229 and (D) MCF-7. * P

    Journal: PLoS ONE

    Article Title: Effect of Melatonin in Epithelial Mesenchymal Transition Markers and Invasive Properties of Breast Cancer Stem Cells of Canine and Human Cell Lines

    doi: 10.1371/journal.pone.0150407

    Figure Lengend Snippet: Graphical representation demonstrating the flow cytometry percentage for CD44/CD24 cancer stem cell phenotype in cells. Flow citometry for cells grown in mololayer for control group of (A) CMTU-229 cell line and (B) MCF-7 cell line. Flow citometry for mammospheres for both cell lines (C) CMTU-229 and (D) MCF-7. * P

    Article Snippet: Flow cytometry analysis By using a Guava easyCyte flow cytometer (Millipore), the expression of stem cell markers in a breast cancer panel was distinctly evaluated in cells from mammospheres.

    Techniques: Flow Cytometry, Cytometry

    Optimization of the operating flow rates within the I-SELEX device to achieve stringent separation of a model non-interacting aptamer from t RBCs while maximizing t RBC recovery. ( a ) Average fluorescence intensity line scans showing the normalized distribution of unbound FITC-labeled CRP aptamers (200 nM) across the channel width at increasing flow rates. Approximate positions of the product and waste outlets are indicated. Corresponding fluorescence images illustrating flow positions of unbound aptamers are also shown as an inset (yellow dashed lines indicate the approximate position of the microchannel walls). ( b ) Average composite images indicate unbound aptamers move to the outer wall and are diverted into the waste outer outlet. ( c ) Average composite images indicate efficient t RBC focusing to the inner microchannel wall and diversion into the product inner outlet. In both ( b ) and ( c ), the sample input and sheath buffer flow rates are 150 μL min −1 and 1500 μL min −1 , respectively, and the yellow dashed lines indicate approximate positions of the microchannel walls and bifurcation. ( d ) Recovery of t RBCs at the product outlet as a percentage of the cells loaded into the device when operated at different sample input flow rates. In all cases, the sheath buffer flow rate is 10-fold higher than the sample input flow rate.

    Journal: Scientific Reports

    Article Title: Identification of malaria parasite-infected red blood cell surface aptamers by inertial microfluidic SELEX (I-SELEX)

    doi: 10.1038/srep11347

    Figure Lengend Snippet: Optimization of the operating flow rates within the I-SELEX device to achieve stringent separation of a model non-interacting aptamer from t RBCs while maximizing t RBC recovery. ( a ) Average fluorescence intensity line scans showing the normalized distribution of unbound FITC-labeled CRP aptamers (200 nM) across the channel width at increasing flow rates. Approximate positions of the product and waste outlets are indicated. Corresponding fluorescence images illustrating flow positions of unbound aptamers are also shown as an inset (yellow dashed lines indicate the approximate position of the microchannel walls). ( b ) Average composite images indicate unbound aptamers move to the outer wall and are diverted into the waste outer outlet. ( c ) Average composite images indicate efficient t RBC focusing to the inner microchannel wall and diversion into the product inner outlet. In both ( b ) and ( c ), the sample input and sheath buffer flow rates are 150 μL min −1 and 1500 μL min −1 , respectively, and the yellow dashed lines indicate approximate positions of the microchannel walls and bifurcation. ( d ) Recovery of t RBCs at the product outlet as a percentage of the cells loaded into the device when operated at different sample input flow rates. In all cases, the sheath buffer flow rate is 10-fold higher than the sample input flow rate.

    Article Snippet: Partitioning efficiency and enrichment experiments For partition efficiency experiments, 100 nM of either Toggle-25 or scr Toggle-25 was incubated in 1 mL TBB with 107 t RBCs for 20 minutes at room temperature with continuous gentle inversion before being passed through the I-SELEX device. t RBCs were recovered directly from the sample outlet onto a vacuum filter plate membrane (Millipore MSHVS4510).

    Techniques: Flow Cytometry, Fluorescence, Labeling

    The I-SELEX device can be used for de novo discovery of high affinity aptamers. ( a ) Bio-layer interferometry kinetic binding data for the interaction between probe-immobilized thrombin and RNA pools from indicated selections rounds. Toggle-25 and scr Toggle-25 were included as positive and negative controls, respectively. ( b ) Comparison of fluorescently labeled 5–12, Toggle-25, scr Toggle-25 and capture oligonucleotide binding to t RBCs versus s RBCs. ( c ) A conserved motif present in Round 3 and 5 selected pools is indicated, and its location within a stem-loop element within the secondary structure predicted for aptamer 5–12 is indicated. ( d ) Evaluation of the importance of this conserved motif to 5–12 thrombin binding affinity was assessed by measuring thrombin binding affinity of: (i) 5–12 mini , in which the 5–12 was truncated while preserving the motif; and (ii) 5–12 mut , in which the motif was mutated in the context of full-length aptamer.

    Journal: Scientific Reports

    Article Title: Identification of malaria parasite-infected red blood cell surface aptamers by inertial microfluidic SELEX (I-SELEX)

    doi: 10.1038/srep11347

    Figure Lengend Snippet: The I-SELEX device can be used for de novo discovery of high affinity aptamers. ( a ) Bio-layer interferometry kinetic binding data for the interaction between probe-immobilized thrombin and RNA pools from indicated selections rounds. Toggle-25 and scr Toggle-25 were included as positive and negative controls, respectively. ( b ) Comparison of fluorescently labeled 5–12, Toggle-25, scr Toggle-25 and capture oligonucleotide binding to t RBCs versus s RBCs. ( c ) A conserved motif present in Round 3 and 5 selected pools is indicated, and its location within a stem-loop element within the secondary structure predicted for aptamer 5–12 is indicated. ( d ) Evaluation of the importance of this conserved motif to 5–12 thrombin binding affinity was assessed by measuring thrombin binding affinity of: (i) 5–12 mini , in which the 5–12 was truncated while preserving the motif; and (ii) 5–12 mut , in which the motif was mutated in the context of full-length aptamer.

    Article Snippet: Partitioning efficiency and enrichment experiments For partition efficiency experiments, 100 nM of either Toggle-25 or scr Toggle-25 was incubated in 1 mL TBB with 107 t RBCs for 20 minutes at room temperature with continuous gentle inversion before being passed through the I-SELEX device. t RBCs were recovered directly from the sample outlet onto a vacuum filter plate membrane (Millipore MSHVS4510).

    Techniques: Binding Assay, Labeling, Preserving

    The I-SELEX device exhibits high partitioning efficiency and can be used to selectively recover and enrich target aptamers from mock libraries. ( a ) Comparison of the recovery of Toggle-25 versus scr Toggle-25 bound to t RBCs targets. ( b ) Enrichment of Toggle-25 aptamers from mock SELEX libraries containing 1:1000 and 1:10 mixtures of Toggle-25: scr Toggle-25 using t RBC targets during a single pass through the I-SELEX device.

    Journal: Scientific Reports

    Article Title: Identification of malaria parasite-infected red blood cell surface aptamers by inertial microfluidic SELEX (I-SELEX)

    doi: 10.1038/srep11347

    Figure Lengend Snippet: The I-SELEX device exhibits high partitioning efficiency and can be used to selectively recover and enrich target aptamers from mock libraries. ( a ) Comparison of the recovery of Toggle-25 versus scr Toggle-25 bound to t RBCs targets. ( b ) Enrichment of Toggle-25 aptamers from mock SELEX libraries containing 1:1000 and 1:10 mixtures of Toggle-25: scr Toggle-25 using t RBC targets during a single pass through the I-SELEX device.

    Article Snippet: Partitioning efficiency and enrichment experiments For partition efficiency experiments, 100 nM of either Toggle-25 or scr Toggle-25 was incubated in 1 mL TBB with 107 t RBCs for 20 minutes at room temperature with continuous gentle inversion before being passed through the I-SELEX device. t RBCs were recovered directly from the sample outlet onto a vacuum filter plate membrane (Millipore MSHVS4510).

    Techniques:

    Characterization of aptamers against malaria-infected red blood cells generated via I-SELEX. ( a ) Relative binding of aptamers isolated from Round 8 of selection 1 (8.1-x) or selection 2 (8.2-x) to CS2 infected RBCs versus uninfected, mock-cultured RBCs. This is plotted as the amount of aptamer recovered for the particular cell type at the sample outlet of the I-SELEX device. ( b ) Binding isotherms for aptamers 8.1-1 and 8.2-1 to CS2 infected RBCs (magenta line), with non-specific binding to mock cultured, uninfected RBCs indicated (black dotted line). ( c ) Glycophorin A (GPA) levels present on the surface of untreated or protease treated RBCs, and Giemsa-stained light microscopy images of the corresponding cells. Scale bar: 8 μm. ( d ) Profiling of aptamers 8.1-1 and 8.2-1 against CS2 infected RBCs (protease treated or untreated) and the DC-J parasite lines that do not express PfEMP1 surface proteins.

    Journal: Scientific Reports

    Article Title: Identification of malaria parasite-infected red blood cell surface aptamers by inertial microfluidic SELEX (I-SELEX)

    doi: 10.1038/srep11347

    Figure Lengend Snippet: Characterization of aptamers against malaria-infected red blood cells generated via I-SELEX. ( a ) Relative binding of aptamers isolated from Round 8 of selection 1 (8.1-x) or selection 2 (8.2-x) to CS2 infected RBCs versus uninfected, mock-cultured RBCs. This is plotted as the amount of aptamer recovered for the particular cell type at the sample outlet of the I-SELEX device. ( b ) Binding isotherms for aptamers 8.1-1 and 8.2-1 to CS2 infected RBCs (magenta line), with non-specific binding to mock cultured, uninfected RBCs indicated (black dotted line). ( c ) Glycophorin A (GPA) levels present on the surface of untreated or protease treated RBCs, and Giemsa-stained light microscopy images of the corresponding cells. Scale bar: 8 μm. ( d ) Profiling of aptamers 8.1-1 and 8.2-1 against CS2 infected RBCs (protease treated or untreated) and the DC-J parasite lines that do not express PfEMP1 surface proteins.

    Article Snippet: Partitioning efficiency and enrichment experiments For partition efficiency experiments, 100 nM of either Toggle-25 or scr Toggle-25 was incubated in 1 mL TBB with 107 t RBCs for 20 minutes at room temperature with continuous gentle inversion before being passed through the I-SELEX device. t RBCs were recovered directly from the sample outlet onto a vacuum filter plate membrane (Millipore MSHVS4510).

    Techniques: Infection, Generated, Binding Assay, Isolation, Selection, Cell Culture, Staining, Light Microscopy

    The I-SELEX microfluidic device and its use in aptamer selection is schematically illustrated. ( a ) The microchannel design consists of a bi-loop spiral of radius ~1 cm with dual inlets and outlets. Pre-incubated bead/cell target-aptamer library mixtures and sheath buffer are pumped through the inner and outer inlets of the device, respectively. Under the influence of Dean drag forces (F D ), unbound aptamers migrate along Dean vortices towards the outer wall and are diverted to the waste outer outlet. Target beads/cells (and any bound aptamers) experience additional strong inertial lift forces (F L ) and are focused along the inner microchannel wall and collected in the product inner outlet. ( b ) A schematic of the I-SELEX procedure showing: (1) negative selection of random library against s RBCs; (2) positive selection of surviving library on t RBCs; (3) partitioning of t RBC-bound aptamers from unbound library in the I-SELEX device; (4) recovery, RT-PCR amplification and in vitro transcription to enrich t RBC-bound aptamers. ( c ) Optical cross sections through the device were taken at positions 1–5 along the length of the channel using high-speed confocal microscopy. Fluorescently labeled CRP aptamers injected via the sample inlet enter the microchannel nearest the inner wall (positions 1 and 2). Aptamers injected via the sample inlet are pinched tightly at the inner wall (position 1). Free aptamers begin to migrate across the channel as a tight band along the midline (positions 2-4) and exit the device when they reach the outer wall (position 5).

    Journal: Scientific Reports

    Article Title: Identification of malaria parasite-infected red blood cell surface aptamers by inertial microfluidic SELEX (I-SELEX)

    doi: 10.1038/srep11347

    Figure Lengend Snippet: The I-SELEX microfluidic device and its use in aptamer selection is schematically illustrated. ( a ) The microchannel design consists of a bi-loop spiral of radius ~1 cm with dual inlets and outlets. Pre-incubated bead/cell target-aptamer library mixtures and sheath buffer are pumped through the inner and outer inlets of the device, respectively. Under the influence of Dean drag forces (F D ), unbound aptamers migrate along Dean vortices towards the outer wall and are diverted to the waste outer outlet. Target beads/cells (and any bound aptamers) experience additional strong inertial lift forces (F L ) and are focused along the inner microchannel wall and collected in the product inner outlet. ( b ) A schematic of the I-SELEX procedure showing: (1) negative selection of random library against s RBCs; (2) positive selection of surviving library on t RBCs; (3) partitioning of t RBC-bound aptamers from unbound library in the I-SELEX device; (4) recovery, RT-PCR amplification and in vitro transcription to enrich t RBC-bound aptamers. ( c ) Optical cross sections through the device were taken at positions 1–5 along the length of the channel using high-speed confocal microscopy. Fluorescently labeled CRP aptamers injected via the sample inlet enter the microchannel nearest the inner wall (positions 1 and 2). Aptamers injected via the sample inlet are pinched tightly at the inner wall (position 1). Free aptamers begin to migrate across the channel as a tight band along the midline (positions 2-4) and exit the device when they reach the outer wall (position 5).

    Article Snippet: Partitioning efficiency and enrichment experiments For partition efficiency experiments, 100 nM of either Toggle-25 or scr Toggle-25 was incubated in 1 mL TBB with 107 t RBCs for 20 minutes at room temperature with continuous gentle inversion before being passed through the I-SELEX device. t RBCs were recovered directly from the sample outlet onto a vacuum filter plate membrane (Millipore MSHVS4510).

    Techniques: Selection, Incubation, Reverse Transcription Polymerase Chain Reaction, Amplification, In Vitro, Confocal Microscopy, Labeling, Injection