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Becton Dickinson facsaria fusion cell sorter
iRFP720 fluorescence and characterisation of pROSA-iRFP720-expressing ID8 cells. ( A ) Fluorescence-activated cell sorting (FACS) enrichment of iRFP720-expressing ID8 cells. iRFP720-positive single cells ([R]730/45 vs. [R]670/30) were sorted into a 96-well plate using the <t>FACSAria</t> Fusion cell sorter (BD Biosciences, San Jose, CA, USA), and single clones were expanded and screened. ( B ) iRFP720 fluorescence was observed using the Cytation 3 imaging Multi-Mode Reader equipped with a Cy5.5 filterset (BioTek Instruments Inc., Winooski, VT, USA). Hoechst 33342 was used to visualise the nucleus (377 ex /447 em ). ( C ) Schematic diagram representing the genomic screening strategy used for identifying iRFP720 incorporation into the ROSA26 locus. ( D ) Genomic DNA isolated from pROSA-puro-iRFP720-transfected single-cell ID8 clones was PCR-amplified using a ROSA26 upstream F primer, an endogenous ROSA26 -specific R primer and an iRFP720-specific R primer, and PCR products were separated with a 1.2% agarose gel. Wild-type ROSA26 gives an 869bp product and successful iRFP720 incorporation gives a 2883bp product. C1–5: clones 1–5; WT: wild-type ID8; NTC: no-template control. ( E ) Proliferation was assessed by electrode impedance using xCELLigence real-time cell analysis. Cells (8 × 10 3 /well) were treated with cisplatin (10 µg/mL) or paclitaxel (20 nM) after 8 h, and proliferation was analysed for further 64 h. ( F ) Wild-type and pROSA-iRFP720 ID8 (3 × 10 5 cells/well) were cultured for 24 h, and then incubated for further 24 h in the presence of cisplatin (10 µg/mL) or paclitaxel (20 nM). Apoptosis was determined by Alexa Fluor®647 annexin V and PI staining on the BD LSRFortessa X-20 (BD Biosciences) flow cytometer. ( G ) Representative images of the apoptosis analysis. Data are presented as mean ± SD, n = 3.
Facsaria Fusion Cell Sorter, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/facsaria fusion cell sorter/product/Becton Dickinson
Average 99 stars, based on 1 article reviews
Price from $9.99 to $1999.99
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1) Product Images from "Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model"

Article Title: Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model

Journal: Cancers

doi: 10.3390/cancers11010032

iRFP720 fluorescence and characterisation of pROSA-iRFP720-expressing ID8 cells. ( A ) Fluorescence-activated cell sorting (FACS) enrichment of iRFP720-expressing ID8 cells. iRFP720-positive single cells ([R]730/45 vs. [R]670/30) were sorted into a 96-well plate using the FACSAria Fusion cell sorter (BD Biosciences, San Jose, CA, USA), and single clones were expanded and screened. ( B ) iRFP720 fluorescence was observed using the Cytation 3 imaging Multi-Mode Reader equipped with a Cy5.5 filterset (BioTek Instruments Inc., Winooski, VT, USA). Hoechst 33342 was used to visualise the nucleus (377 ex /447 em ). ( C ) Schematic diagram representing the genomic screening strategy used for identifying iRFP720 incorporation into the ROSA26 locus. ( D ) Genomic DNA isolated from pROSA-puro-iRFP720-transfected single-cell ID8 clones was PCR-amplified using a ROSA26 upstream F primer, an endogenous ROSA26 -specific R primer and an iRFP720-specific R primer, and PCR products were separated with a 1.2% agarose gel. Wild-type ROSA26 gives an 869bp product and successful iRFP720 incorporation gives a 2883bp product. C1–5: clones 1–5; WT: wild-type ID8; NTC: no-template control. ( E ) Proliferation was assessed by electrode impedance using xCELLigence real-time cell analysis. Cells (8 × 10 3 /well) were treated with cisplatin (10 µg/mL) or paclitaxel (20 nM) after 8 h, and proliferation was analysed for further 64 h. ( F ) Wild-type and pROSA-iRFP720 ID8 (3 × 10 5 cells/well) were cultured for 24 h, and then incubated for further 24 h in the presence of cisplatin (10 µg/mL) or paclitaxel (20 nM). Apoptosis was determined by Alexa Fluor®647 annexin V and PI staining on the BD LSRFortessa X-20 (BD Biosciences) flow cytometer. ( G ) Representative images of the apoptosis analysis. Data are presented as mean ± SD, n = 3.
Figure Legend Snippet: iRFP720 fluorescence and characterisation of pROSA-iRFP720-expressing ID8 cells. ( A ) Fluorescence-activated cell sorting (FACS) enrichment of iRFP720-expressing ID8 cells. iRFP720-positive single cells ([R]730/45 vs. [R]670/30) were sorted into a 96-well plate using the FACSAria Fusion cell sorter (BD Biosciences, San Jose, CA, USA), and single clones were expanded and screened. ( B ) iRFP720 fluorescence was observed using the Cytation 3 imaging Multi-Mode Reader equipped with a Cy5.5 filterset (BioTek Instruments Inc., Winooski, VT, USA). Hoechst 33342 was used to visualise the nucleus (377 ex /447 em ). ( C ) Schematic diagram representing the genomic screening strategy used for identifying iRFP720 incorporation into the ROSA26 locus. ( D ) Genomic DNA isolated from pROSA-puro-iRFP720-transfected single-cell ID8 clones was PCR-amplified using a ROSA26 upstream F primer, an endogenous ROSA26 -specific R primer and an iRFP720-specific R primer, and PCR products were separated with a 1.2% agarose gel. Wild-type ROSA26 gives an 869bp product and successful iRFP720 incorporation gives a 2883bp product. C1–5: clones 1–5; WT: wild-type ID8; NTC: no-template control. ( E ) Proliferation was assessed by electrode impedance using xCELLigence real-time cell analysis. Cells (8 × 10 3 /well) were treated with cisplatin (10 µg/mL) or paclitaxel (20 nM) after 8 h, and proliferation was analysed for further 64 h. ( F ) Wild-type and pROSA-iRFP720 ID8 (3 × 10 5 cells/well) were cultured for 24 h, and then incubated for further 24 h in the presence of cisplatin (10 µg/mL) or paclitaxel (20 nM). Apoptosis was determined by Alexa Fluor®647 annexin V and PI staining on the BD LSRFortessa X-20 (BD Biosciences) flow cytometer. ( G ) Representative images of the apoptosis analysis. Data are presented as mean ± SD, n = 3.

Techniques Used: Fluorescence, Expressing, FACS, Clone Assay, Imaging, Isolation, Transfection, Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Cell Culture, Incubation, Staining, Flow Cytometry

Related Articles

FACS:

Article Title: Identification of an Aptamer Binding to Human Osteogenic-Induced Progenitor Cells
Article Snippet: However, the cells were resuspended in PBS instead of FACS buffer to maintain cell viability and physiological conditions for further cultivation after the cell sorting. .. The cells were sorted using a BD FACS Aria cell sorter (Becton Dickinson), and only the strongly aptamer 74-stained cells and the strongly negative fraction were plated for the analyses of the mineralizing capacity and gene expression. .. Immediately after cell sorting, the aptamer 74-positive and -negative cells were washed with PBS, centrifuged and plated on special E-plates (Roche) at a cell density of 5×102 cells per well.

Article Title: Identification of PMN-released mutagenic factors in a co-culture model for colitis-associated cancer
Article Snippet: PMNs were activated with 0.5 nM PMA, if not stated otherwise, in cell culture medium. .. Cell sorting, co-culture, flow cytometry, analysis of mutations and calculation of mutation rates 5 × 103 –1 × 104 EGFP-negative HCT116 + chr3 or HCEC-1CT frameshift-reporter cells were sorted into 24-well plates on a FACSAria cell sorter using CloneCyt Plus software (BD Biocsiences, San Jose, CA). .. Twenty-four hours later, freshly isolated PMNs were added together with 0.5 nM PMA at effector:target ratios ranging from 0:1 (control) to 75:1.

Article Title: De Novo Synthesis of Phosphatidylcholine Is Essential for the Promastigote But Not Amastigote Stage in Leishmania major
Article Snippet: For every passage, the GFP expression level was determined by flow cytometry at mid-log phase (3–7 × 106 cells/ml) using an Attune NxT Acoustic Flow Cytometer. .. After 16 passages in the presence of 50 μg/ml of GCV, cept — + pXNG4-CEPT parasites were sorted based on their GFP expression levels using a BD Biosciences FACS Aria III Plus Cell Sorter. ..

Article Title: Interleukin-33 and thymic stromal lymphopoietin, but not interleukin-25, are crucial for development of airway eosinophilia induced by chitin
Article Snippet: The cells in the negative fraction were collected and then incubated (30 min on ice) with a mixture containing PerCP/Cy5.5-conjugated anti-mouse KLRG1(Killer cell Lectin-like Receptor G1) mAb (2F1/KLRG1; BioLegend), PE/Cy7-conjugated anti-mouse CD127 mAb (A7R34; BioLegend), APC-conjugated anti-mouse Sca-1 mAb (D7; BioLegend), BD Horizon BV510-conjugated anti-mouse CD45 mAb (30-F11; BioLegend), BV421-conjugated Streptavidin (BioLegend) and eBioscience Fixable Viability Dye eFluor 780 (Thermo Fisher Scientific). .. After washing, Viability Dye-negative CD45+ lineage− Sca-1+ KLRG1+ CD127+ cells, i.e., ILC2s, were sorted using a FACS Aria II Cell Sorter (BD Biosciences) with BD FACSDiva v6.1.3 software (BD Biosciences). .. The ILC2s were resuspended in 200 μl of RPMI1640 medium supplemented with 10% FBS, 100 U/ml penicillin and 100 μg/ml streptomycin, and then cultured in the presence and absence of 50 ng/ml of each of recombinant mouse IL-25 (R & D Systems), recombinant mouse IL-33 (R & D Systems) and recombinant mouse TSLP (R & D Systems) in 96-well round-bottomed plates (680–1000 cells/well) at 37 °C in a 5.0% CO2 incubator for 2 days.

Article Title: Transcriptional Profiling Reveals Crosstalk Between Mesenchymal Stem Cells and Endothelial Cells Promoting Prevascularization by Reciprocal Mechanisms
Article Snippet: During fluorescence activated cell sortor (FACS)-based cell sorting, cells from each group were trypsinized and washed thrice with phosphate buffer solution (PBS). .. GFP-positive HMSC populations were immediately isolated with a BD FACS cell sorter (Aril III; BD Biosciences). .. During CD31+ cell sorting, the GFP-negative populations were incubated with Cy3-conjugated anti-CD31 antibody (anti-CD31-Cy3; Sino Biologicals) and sorted with BD FACS.

Article Title: Head and Neck Squamous Cell Carcinoma Metabolism Draws on Glutaminolysis, and Stemness Is Specifically Regulated by Glutaminolysis via Aldehyde Dehydrogenase
Article Snippet: Cells were costained with an APC antihuman CD44 antibody (559942, BD Pharmingen, San Diego, CA). .. The expression of CD44 and ALDH was analyzed by flow cytometry using a FACS Aria Cell Sorter (BD Biosciences, San Jose, CA). .. For tumorsphere assays, ALDHhi CD44hi and ALDHlo CD44lo cells, derived from FACS, were plated on poly-HEMA coated plates at a density of 5000 viable cells/well.

Expressing:

Article Title: Identification of an Aptamer Binding to Human Osteogenic-Induced Progenitor Cells
Article Snippet: However, the cells were resuspended in PBS instead of FACS buffer to maintain cell viability and physiological conditions for further cultivation after the cell sorting. .. The cells were sorted using a BD FACS Aria cell sorter (Becton Dickinson), and only the strongly aptamer 74-stained cells and the strongly negative fraction were plated for the analyses of the mineralizing capacity and gene expression. .. Immediately after cell sorting, the aptamer 74-positive and -negative cells were washed with PBS, centrifuged and plated on special E-plates (Roche) at a cell density of 5×102 cells per well.

Article Title: De Novo Synthesis of Phosphatidylcholine Is Essential for the Promastigote But Not Amastigote Stage in Leishmania major
Article Snippet: For every passage, the GFP expression level was determined by flow cytometry at mid-log phase (3–7 × 106 cells/ml) using an Attune NxT Acoustic Flow Cytometer. .. After 16 passages in the presence of 50 μg/ml of GCV, cept — + pXNG4-CEPT parasites were sorted based on their GFP expression levels using a BD Biosciences FACS Aria III Plus Cell Sorter. ..

Article Title: Head and Neck Squamous Cell Carcinoma Metabolism Draws on Glutaminolysis, and Stemness Is Specifically Regulated by Glutaminolysis via Aldehyde Dehydrogenase
Article Snippet: Cells were costained with an APC antihuman CD44 antibody (559942, BD Pharmingen, San Diego, CA). .. The expression of CD44 and ALDH was analyzed by flow cytometry using a FACS Aria Cell Sorter (BD Biosciences, San Jose, CA). .. For tumorsphere assays, ALDHhi CD44hi and ALDHlo CD44lo cells, derived from FACS, were plated on poly-HEMA coated plates at a density of 5000 viable cells/well.

Co-Culture Assay:

Article Title: Identification of PMN-released mutagenic factors in a co-culture model for colitis-associated cancer
Article Snippet: PMNs were activated with 0.5 nM PMA, if not stated otherwise, in cell culture medium. .. Cell sorting, co-culture, flow cytometry, analysis of mutations and calculation of mutation rates 5 × 103 –1 × 104 EGFP-negative HCT116 + chr3 or HCEC-1CT frameshift-reporter cells were sorted into 24-well plates on a FACSAria cell sorter using CloneCyt Plus software (BD Biocsiences, San Jose, CA). .. Twenty-four hours later, freshly isolated PMNs were added together with 0.5 nM PMA at effector:target ratios ranging from 0:1 (control) to 75:1.

Flow Cytometry:

Article Title: Identification of PMN-released mutagenic factors in a co-culture model for colitis-associated cancer
Article Snippet: PMNs were activated with 0.5 nM PMA, if not stated otherwise, in cell culture medium. .. Cell sorting, co-culture, flow cytometry, analysis of mutations and calculation of mutation rates 5 × 103 –1 × 104 EGFP-negative HCT116 + chr3 or HCEC-1CT frameshift-reporter cells were sorted into 24-well plates on a FACSAria cell sorter using CloneCyt Plus software (BD Biocsiences, San Jose, CA). .. Twenty-four hours later, freshly isolated PMNs were added together with 0.5 nM PMA at effector:target ratios ranging from 0:1 (control) to 75:1.

Article Title: Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model
Article Snippet: ID8 cells were grown to 70–80% confluence and then co-transfected with pROSA-puro-iRFP720 and plentiCRISPRv1 ROSA26-sgRNA2 or ROSA26-sgRNA4 using Lipofectamine 2000 (Invitrogen, CA, USA), using 3.5 µg of DNA in total. .. Transfected ID8 cells were maintained in puromycin selection for two weeks and then sorted by flow cytometry for single iRFP720+ clones ([R]730/45 vs. [R]670/30) using the FACSAria Fusion cell sorter (BD Biosciences, San Jose, CA, USA). .. Clones were maintained under selective pressure in DMEM/FBS/ITS/PS until outgrowth was evident.

Article Title: Head and Neck Squamous Cell Carcinoma Metabolism Draws on Glutaminolysis, and Stemness Is Specifically Regulated by Glutaminolysis via Aldehyde Dehydrogenase
Article Snippet: Cells were costained with an APC antihuman CD44 antibody (559942, BD Pharmingen, San Diego, CA). .. The expression of CD44 and ALDH was analyzed by flow cytometry using a FACS Aria Cell Sorter (BD Biosciences, San Jose, CA). .. For tumorsphere assays, ALDHhi CD44hi and ALDHlo CD44lo cells, derived from FACS, were plated on poly-HEMA coated plates at a density of 5000 viable cells/well.

Mutagenesis:

Article Title: Identification of PMN-released mutagenic factors in a co-culture model for colitis-associated cancer
Article Snippet: PMNs were activated with 0.5 nM PMA, if not stated otherwise, in cell culture medium. .. Cell sorting, co-culture, flow cytometry, analysis of mutations and calculation of mutation rates 5 × 103 –1 × 104 EGFP-negative HCT116 + chr3 or HCEC-1CT frameshift-reporter cells were sorted into 24-well plates on a FACSAria cell sorter using CloneCyt Plus software (BD Biocsiences, San Jose, CA). .. Twenty-four hours later, freshly isolated PMNs were added together with 0.5 nM PMA at effector:target ratios ranging from 0:1 (control) to 75:1.

Software:

Article Title: Identification of PMN-released mutagenic factors in a co-culture model for colitis-associated cancer
Article Snippet: PMNs were activated with 0.5 nM PMA, if not stated otherwise, in cell culture medium. .. Cell sorting, co-culture, flow cytometry, analysis of mutations and calculation of mutation rates 5 × 103 –1 × 104 EGFP-negative HCT116 + chr3 or HCEC-1CT frameshift-reporter cells were sorted into 24-well plates on a FACSAria cell sorter using CloneCyt Plus software (BD Biocsiences, San Jose, CA). .. Twenty-four hours later, freshly isolated PMNs were added together with 0.5 nM PMA at effector:target ratios ranging from 0:1 (control) to 75:1.

Article Title: Interleukin-33 and thymic stromal lymphopoietin, but not interleukin-25, are crucial for development of airway eosinophilia induced by chitin
Article Snippet: The cells in the negative fraction were collected and then incubated (30 min on ice) with a mixture containing PerCP/Cy5.5-conjugated anti-mouse KLRG1(Killer cell Lectin-like Receptor G1) mAb (2F1/KLRG1; BioLegend), PE/Cy7-conjugated anti-mouse CD127 mAb (A7R34; BioLegend), APC-conjugated anti-mouse Sca-1 mAb (D7; BioLegend), BD Horizon BV510-conjugated anti-mouse CD45 mAb (30-F11; BioLegend), BV421-conjugated Streptavidin (BioLegend) and eBioscience Fixable Viability Dye eFluor 780 (Thermo Fisher Scientific). .. After washing, Viability Dye-negative CD45+ lineage− Sca-1+ KLRG1+ CD127+ cells, i.e., ILC2s, were sorted using a FACS Aria II Cell Sorter (BD Biosciences) with BD FACSDiva v6.1.3 software (BD Biosciences). .. The ILC2s were resuspended in 200 μl of RPMI1640 medium supplemented with 10% FBS, 100 U/ml penicillin and 100 μg/ml streptomycin, and then cultured in the presence and absence of 50 ng/ml of each of recombinant mouse IL-25 (R & D Systems), recombinant mouse IL-33 (R & D Systems) and recombinant mouse TSLP (R & D Systems) in 96-well round-bottomed plates (680–1000 cells/well) at 37 °C in a 5.0% CO2 incubator for 2 days.

Transfection:

Article Title: Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model
Article Snippet: ID8 cells were grown to 70–80% confluence and then co-transfected with pROSA-puro-iRFP720 and plentiCRISPRv1 ROSA26-sgRNA2 or ROSA26-sgRNA4 using Lipofectamine 2000 (Invitrogen, CA, USA), using 3.5 µg of DNA in total. .. Transfected ID8 cells were maintained in puromycin selection for two weeks and then sorted by flow cytometry for single iRFP720+ clones ([R]730/45 vs. [R]670/30) using the FACSAria Fusion cell sorter (BD Biosciences, San Jose, CA, USA). .. Clones were maintained under selective pressure in DMEM/FBS/ITS/PS until outgrowth was evident.

Selection:

Article Title: Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model
Article Snippet: ID8 cells were grown to 70–80% confluence and then co-transfected with pROSA-puro-iRFP720 and plentiCRISPRv1 ROSA26-sgRNA2 or ROSA26-sgRNA4 using Lipofectamine 2000 (Invitrogen, CA, USA), using 3.5 µg of DNA in total. .. Transfected ID8 cells were maintained in puromycin selection for two weeks and then sorted by flow cytometry for single iRFP720+ clones ([R]730/45 vs. [R]670/30) using the FACSAria Fusion cell sorter (BD Biosciences, San Jose, CA, USA). .. Clones were maintained under selective pressure in DMEM/FBS/ITS/PS until outgrowth was evident.

Clone Assay:

Article Title: Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model
Article Snippet: ID8 cells were grown to 70–80% confluence and then co-transfected with pROSA-puro-iRFP720 and plentiCRISPRv1 ROSA26-sgRNA2 or ROSA26-sgRNA4 using Lipofectamine 2000 (Invitrogen, CA, USA), using 3.5 µg of DNA in total. .. Transfected ID8 cells were maintained in puromycin selection for two weeks and then sorted by flow cytometry for single iRFP720+ clones ([R]730/45 vs. [R]670/30) using the FACSAria Fusion cell sorter (BD Biosciences, San Jose, CA, USA). .. Clones were maintained under selective pressure in DMEM/FBS/ITS/PS until outgrowth was evident.

Isolation:

Article Title: Transcriptional Profiling Reveals Crosstalk Between Mesenchymal Stem Cells and Endothelial Cells Promoting Prevascularization by Reciprocal Mechanisms
Article Snippet: During fluorescence activated cell sortor (FACS)-based cell sorting, cells from each group were trypsinized and washed thrice with phosphate buffer solution (PBS). .. GFP-positive HMSC populations were immediately isolated with a BD FACS cell sorter (Aril III; BD Biosciences). .. During CD31+ cell sorting, the GFP-negative populations were incubated with Cy3-conjugated anti-CD31 antibody (anti-CD31-Cy3; Sino Biologicals) and sorted with BD FACS.

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    Becton Dickinson facsaria fusion cell sorter
    iRFP720 fluorescence and characterisation of pROSA-iRFP720-expressing ID8 cells. ( A ) Fluorescence-activated cell sorting (FACS) enrichment of iRFP720-expressing ID8 cells. iRFP720-positive single cells ([R]730/45 vs. [R]670/30) were sorted into a 96-well plate using the <t>FACSAria</t> Fusion cell sorter (BD Biosciences, San Jose, CA, USA), and single clones were expanded and screened. ( B ) iRFP720 fluorescence was observed using the Cytation 3 imaging Multi-Mode Reader equipped with a Cy5.5 filterset (BioTek Instruments Inc., Winooski, VT, USA). Hoechst 33342 was used to visualise the nucleus (377 ex /447 em ). ( C ) Schematic diagram representing the genomic screening strategy used for identifying iRFP720 incorporation into the ROSA26 locus. ( D ) Genomic DNA isolated from pROSA-puro-iRFP720-transfected single-cell ID8 clones was PCR-amplified using a ROSA26 upstream F primer, an endogenous ROSA26 -specific R primer and an iRFP720-specific R primer, and PCR products were separated with a 1.2% agarose gel. Wild-type ROSA26 gives an 869bp product and successful iRFP720 incorporation gives a 2883bp product. C1–5: clones 1–5; WT: wild-type ID8; NTC: no-template control. ( E ) Proliferation was assessed by electrode impedance using xCELLigence real-time cell analysis. Cells (8 × 10 3 /well) were treated with cisplatin (10 µg/mL) or paclitaxel (20 nM) after 8 h, and proliferation was analysed for further 64 h. ( F ) Wild-type and pROSA-iRFP720 ID8 (3 × 10 5 cells/well) were cultured for 24 h, and then incubated for further 24 h in the presence of cisplatin (10 µg/mL) or paclitaxel (20 nM). Apoptosis was determined by Alexa Fluor®647 annexin V and PI staining on the BD LSRFortessa X-20 (BD Biosciences) flow cytometer. ( G ) Representative images of the apoptosis analysis. Data are presented as mean ± SD, n = 3.
    Facsaria Fusion Cell Sorter, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/facsaria fusion cell sorter/product/Becton Dickinson
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    facsaria fusion cell sorter - by Bioz Stars, 2021-06
    99/100 stars
      Buy from Supplier

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    iRFP720 fluorescence and characterisation of pROSA-iRFP720-expressing ID8 cells. ( A ) Fluorescence-activated cell sorting (FACS) enrichment of iRFP720-expressing ID8 cells. iRFP720-positive single cells ([R]730/45 vs. [R]670/30) were sorted into a 96-well plate using the FACSAria Fusion cell sorter (BD Biosciences, San Jose, CA, USA), and single clones were expanded and screened. ( B ) iRFP720 fluorescence was observed using the Cytation 3 imaging Multi-Mode Reader equipped with a Cy5.5 filterset (BioTek Instruments Inc., Winooski, VT, USA). Hoechst 33342 was used to visualise the nucleus (377 ex /447 em ). ( C ) Schematic diagram representing the genomic screening strategy used for identifying iRFP720 incorporation into the ROSA26 locus. ( D ) Genomic DNA isolated from pROSA-puro-iRFP720-transfected single-cell ID8 clones was PCR-amplified using a ROSA26 upstream F primer, an endogenous ROSA26 -specific R primer and an iRFP720-specific R primer, and PCR products were separated with a 1.2% agarose gel. Wild-type ROSA26 gives an 869bp product and successful iRFP720 incorporation gives a 2883bp product. C1–5: clones 1–5; WT: wild-type ID8; NTC: no-template control. ( E ) Proliferation was assessed by electrode impedance using xCELLigence real-time cell analysis. Cells (8 × 10 3 /well) were treated with cisplatin (10 µg/mL) or paclitaxel (20 nM) after 8 h, and proliferation was analysed for further 64 h. ( F ) Wild-type and pROSA-iRFP720 ID8 (3 × 10 5 cells/well) were cultured for 24 h, and then incubated for further 24 h in the presence of cisplatin (10 µg/mL) or paclitaxel (20 nM). Apoptosis was determined by Alexa Fluor®647 annexin V and PI staining on the BD LSRFortessa X-20 (BD Biosciences) flow cytometer. ( G ) Representative images of the apoptosis analysis. Data are presented as mean ± SD, n = 3.

    Journal: Cancers

    Article Title: Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model

    doi: 10.3390/cancers11010032

    Figure Lengend Snippet: iRFP720 fluorescence and characterisation of pROSA-iRFP720-expressing ID8 cells. ( A ) Fluorescence-activated cell sorting (FACS) enrichment of iRFP720-expressing ID8 cells. iRFP720-positive single cells ([R]730/45 vs. [R]670/30) were sorted into a 96-well plate using the FACSAria Fusion cell sorter (BD Biosciences, San Jose, CA, USA), and single clones were expanded and screened. ( B ) iRFP720 fluorescence was observed using the Cytation 3 imaging Multi-Mode Reader equipped with a Cy5.5 filterset (BioTek Instruments Inc., Winooski, VT, USA). Hoechst 33342 was used to visualise the nucleus (377 ex /447 em ). ( C ) Schematic diagram representing the genomic screening strategy used for identifying iRFP720 incorporation into the ROSA26 locus. ( D ) Genomic DNA isolated from pROSA-puro-iRFP720-transfected single-cell ID8 clones was PCR-amplified using a ROSA26 upstream F primer, an endogenous ROSA26 -specific R primer and an iRFP720-specific R primer, and PCR products were separated with a 1.2% agarose gel. Wild-type ROSA26 gives an 869bp product and successful iRFP720 incorporation gives a 2883bp product. C1–5: clones 1–5; WT: wild-type ID8; NTC: no-template control. ( E ) Proliferation was assessed by electrode impedance using xCELLigence real-time cell analysis. Cells (8 × 10 3 /well) were treated with cisplatin (10 µg/mL) or paclitaxel (20 nM) after 8 h, and proliferation was analysed for further 64 h. ( F ) Wild-type and pROSA-iRFP720 ID8 (3 × 10 5 cells/well) were cultured for 24 h, and then incubated for further 24 h in the presence of cisplatin (10 µg/mL) or paclitaxel (20 nM). Apoptosis was determined by Alexa Fluor®647 annexin V and PI staining on the BD LSRFortessa X-20 (BD Biosciences) flow cytometer. ( G ) Representative images of the apoptosis analysis. Data are presented as mean ± SD, n = 3.

    Article Snippet: Transfected ID8 cells were maintained in puromycin selection for two weeks and then sorted by flow cytometry for single iRFP720+ clones ([R]730/45 vs. [R]670/30) using the FACSAria Fusion cell sorter (BD Biosciences, San Jose, CA, USA).

    Techniques: Fluorescence, Expressing, FACS, Clone Assay, Imaging, Isolation, Transfection, Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Cell Culture, Incubation, Staining, Flow Cytometry

    HNSCC cancer stem cells (CD44 hi /ALDH hi ) and ALDH expression are regulated by glutaminolysis. (A) Representative FACS-based sorting of ALDH hi CD44 hi and ALDH lo CD44 lo fractions from UM-SCC-14A cells stained with the Aldefluor reagent. The nonviable cells

    Journal: Journal of proteome research

    Article Title: Head and Neck Squamous Cell Carcinoma Metabolism Draws on Glutaminolysis, and Stemness Is Specifically Regulated by Glutaminolysis via Aldehyde Dehydrogenase

    doi: 10.1021/acs.jproteome.6b00936

    Figure Lengend Snippet: HNSCC cancer stem cells (CD44 hi /ALDH hi ) and ALDH expression are regulated by glutaminolysis. (A) Representative FACS-based sorting of ALDH hi CD44 hi and ALDH lo CD44 lo fractions from UM-SCC-14A cells stained with the Aldefluor reagent. The nonviable cells

    Article Snippet: The expression of CD44 and ALDH was analyzed by flow cytometry using a FACS Aria Cell Sorter (BD Biosciences, San Jose, CA).

    Techniques: Expressing, FACS, Staining

    Induction of frameshift mutations at mono-, di- and tetranucleotide repeats by co-culture of activated PMNs and HCT116 + chr3 ( A ) or HCEC-1CT ( B ). 5 × 10 3 EGPF-negative reporter cells were sorted on a FACSaria instrument and were incubated with PMA-activated (0.5 nM) PMNs at different effector (PMNs):target (epithelial cells) ratios the following day for 24 h. flow cytometric analysis was performed 6 days later showing a dose-dependent decrease in cell numbers and an increase in MR (presented as fold change to untreated control cells) for all clones. Measurements were carried out in quadruplicates. * P

    Journal: Carcinogenesis

    Article Title: Identification of PMN-released mutagenic factors in a co-culture model for colitis-associated cancer

    doi: 10.1093/carcin/bgx118

    Figure Lengend Snippet: Induction of frameshift mutations at mono-, di- and tetranucleotide repeats by co-culture of activated PMNs and HCT116 + chr3 ( A ) or HCEC-1CT ( B ). 5 × 10 3 EGPF-negative reporter cells were sorted on a FACSaria instrument and were incubated with PMA-activated (0.5 nM) PMNs at different effector (PMNs):target (epithelial cells) ratios the following day for 24 h. flow cytometric analysis was performed 6 days later showing a dose-dependent decrease in cell numbers and an increase in MR (presented as fold change to untreated control cells) for all clones. Measurements were carried out in quadruplicates. * P

    Article Snippet: Cell sorting, co-culture, flow cytometry, analysis of mutations and calculation of mutation rates 5 × 103 –1 × 104 EGFP-negative HCT116 + chr3 or HCEC-1CT frameshift-reporter cells were sorted into 24-well plates on a FACSAria cell sorter using CloneCyt Plus software (BD Biocsiences, San Jose, CA).

    Techniques: Co-Culture Assay, Incubation, Clone Assay