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    Becton Dickinson facsaria iii cell sorter
    HEK293T cells co-expressing the fusion proteins in the following combinations were analyzed with the configured BD <t>FACSAria™</t> <t>III</t> cell sorter. ( A ) BST-2 fused Clover and HIV-2 Env WT fused to mRuby2; ( B ) BST-2 fused Clover and HIV-2 Env 1–749 fused mRuby2; ( C ) BST-2 fused Clover and HIV-2 Env N659D fused mRuby2 and ( D ) BST-2 fused Clover and HIV-2 Nef fused mRuby2. Numbers indicated in gate P4 give the percentage of FRET-positive cells included in P2 according to the previous fluorescence-activated cell sorting (FACS) configuration for the different co-expression in living cells ( Figure 6 ); ( E ) Bar diagram summarizing the FRET-positive signals from four independent experiments ( n = 4). Statistical tests give a significance with a p
    Facsaria Iii Cell Sorter, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Becton Dickinson bd facsaria iii cell sorter
    RNA-seq of stained and sorted cell populations. A : Bioanalyzer profiles of total RNA derived from unprocessed cells (harvested directly from trypsinised cells) compared to cells that have undergone glyoxal fixation, permeabilisation with 100% methanol and indirect immunofluorescence for CCNB1. 0.1–5 ng of total RNA was separated on total RNA pico chips on Agilent 2100 Bioanalyzer. B : Scatter plots comparing the normalised read count for each annotated gene in GRCh38 in poly(A)+ RNA-seq libraries derived from unprocessed and from glyoxal fixed, permeabilised and stained cells. Two biological replicates were sequenced for each condition and averaged, read counts were normalised to the total number of reads in each library. Data is shown for both COLO205 and MCF-7 cells. C : Flow cytometry density plots for MCF-7 cells labelled with anti-CCNB1 primary antibody and donkey Alexa Fluor-488 conjugated secondary antibody and sorted using a BD <t>FACSAria</t> <t>III</t> sorter. Intact cells (shown within the elliptical gate) were distinguished from off-scale events and cell debris using forward scatter (FSC) and side scatter (SSC) measurements (panel 1). Doublets were excluded from the gated cells by a 2-step gating strategy with pulse height (H) plotted against area (A) for FSC parameter followed by SSC-H versus SSC-A plot (panel 3). Fluorescence thresholds for isolation of CCNB1 positive and negative cell fractions are shown in panel 4. Gates were set using the negative control and the CCNB1 positive and negative sorting gates were moved apart from each other to maximise purity when sorted. D : Fluorescence histogram of DAPI intensities for sorted CCNB1 negative and positive populations. E : Scatter plots comparing the normalised read count for each annotated gene in GRCh38 in poly(A)+ RNA-seq libraries derived from two biological replicates each of CCNB1 positive (left) and negative (right) MCF-7 cells. Note the particularly tight correlation after sorting towards the antigen of interest. E : Hierarchical clustering analysis of the 161 significantly different expressed genes identified DEseq2 analysis (p
    Bd Facsaria Iii Cell Sorter, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bd facsaria iii cell sorter/product/Becton Dickinson
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bd facsaria iii cell sorter - by Bioz Stars, 2021-07
    86/100 stars
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    99
    Becton Dickinson facs
    RNA-seq of stained and sorted cell populations. A : Bioanalyzer profiles of total RNA derived from unprocessed cells (harvested directly from trypsinised cells) compared to cells that have undergone glyoxal fixation, permeabilisation with 100% methanol and indirect immunofluorescence for CCNB1. 0.1–5 ng of total RNA was separated on total RNA pico chips on Agilent 2100 Bioanalyzer. B : Scatter plots comparing the normalised read count for each annotated gene in GRCh38 in poly(A)+ RNA-seq libraries derived from unprocessed and from glyoxal fixed, permeabilised and stained cells. Two biological replicates were sequenced for each condition and averaged, read counts were normalised to the total number of reads in each library. Data is shown for both COLO205 and MCF-7 cells. C : Flow cytometry density plots for MCF-7 cells labelled with anti-CCNB1 primary antibody and donkey Alexa Fluor-488 conjugated secondary antibody and sorted using a BD <t>FACSAria</t> <t>III</t> sorter. Intact cells (shown within the elliptical gate) were distinguished from off-scale events and cell debris using forward scatter (FSC) and side scatter (SSC) measurements (panel 1). Doublets were excluded from the gated cells by a 2-step gating strategy with pulse height (H) plotted against area (A) for FSC parameter followed by SSC-H versus SSC-A plot (panel 3). Fluorescence thresholds for isolation of CCNB1 positive and negative cell fractions are shown in panel 4. Gates were set using the negative control and the CCNB1 positive and negative sorting gates were moved apart from each other to maximise purity when sorted. D : Fluorescence histogram of DAPI intensities for sorted CCNB1 negative and positive populations. E : Scatter plots comparing the normalised read count for each annotated gene in GRCh38 in poly(A)+ RNA-seq libraries derived from two biological replicates each of CCNB1 positive (left) and negative (right) MCF-7 cells. Note the particularly tight correlation after sorting towards the antigen of interest. E : Hierarchical clustering analysis of the 161 significantly different expressed genes identified DEseq2 analysis (p
    Facs, 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/facs/product/Becton Dickinson
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    facs - by Bioz Stars, 2021-07
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    86
    Becton Dickinson facsaria iii sorter
    RNA-seq of stained and sorted cell populations. A : Bioanalyzer profiles of total RNA derived from unprocessed cells (harvested directly from trypsinised cells) compared to cells that have undergone glyoxal fixation, permeabilisation with 100% methanol and indirect immunofluorescence for CCNB1. 0.1–5 ng of total RNA was separated on total RNA pico chips on Agilent 2100 Bioanalyzer. B : Scatter plots comparing the normalised read count for each annotated gene in GRCh38 in poly(A)+ RNA-seq libraries derived from unprocessed and from glyoxal fixed, permeabilised and stained cells. Two biological replicates were sequenced for each condition and averaged, read counts were normalised to the total number of reads in each library. Data is shown for both COLO205 and MCF-7 cells. C : Flow cytometry density plots for MCF-7 cells labelled with anti-CCNB1 primary antibody and donkey Alexa Fluor-488 conjugated secondary antibody and sorted using a BD <t>FACSAria</t> <t>III</t> sorter. Intact cells (shown within the elliptical gate) were distinguished from off-scale events and cell debris using forward scatter (FSC) and side scatter (SSC) measurements (panel 1). Doublets were excluded from the gated cells by a 2-step gating strategy with pulse height (H) plotted against area (A) for FSC parameter followed by SSC-H versus SSC-A plot (panel 3). Fluorescence thresholds for isolation of CCNB1 positive and negative cell fractions are shown in panel 4. Gates were set using the negative control and the CCNB1 positive and negative sorting gates were moved apart from each other to maximise purity when sorted. D : Fluorescence histogram of DAPI intensities for sorted CCNB1 negative and positive populations. E : Scatter plots comparing the normalised read count for each annotated gene in GRCh38 in poly(A)+ RNA-seq libraries derived from two biological replicates each of CCNB1 positive (left) and negative (right) MCF-7 cells. Note the particularly tight correlation after sorting towards the antigen of interest. E : Hierarchical clustering analysis of the 161 significantly different expressed genes identified DEseq2 analysis (p
    Facsaria Iii Sorter, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/facsaria iii sorter/product/Becton Dickinson
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    facsaria iii sorter - by Bioz Stars, 2021-07
    86/100 stars
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    Image Search Results


    HEK293T cells co-expressing the fusion proteins in the following combinations were analyzed with the configured BD FACSAria™ III cell sorter. ( A ) BST-2 fused Clover and HIV-2 Env WT fused to mRuby2; ( B ) BST-2 fused Clover and HIV-2 Env 1–749 fused mRuby2; ( C ) BST-2 fused Clover and HIV-2 Env N659D fused mRuby2 and ( D ) BST-2 fused Clover and HIV-2 Nef fused mRuby2. Numbers indicated in gate P4 give the percentage of FRET-positive cells included in P2 according to the previous fluorescence-activated cell sorting (FACS) configuration for the different co-expression in living cells ( Figure 6 ); ( E ) Bar diagram summarizing the FRET-positive signals from four independent experiments ( n = 4). Statistical tests give a significance with a p

    Journal: Viruses

    Article Title: Single Amino Acid Substitution N659D in HIV-2 Envelope Glycoprotein (Env) Impairs Viral Release and Hampers BST-2 Antagonism

    doi: 10.3390/v8100285

    Figure Lengend Snippet: HEK293T cells co-expressing the fusion proteins in the following combinations were analyzed with the configured BD FACSAria™ III cell sorter. ( A ) BST-2 fused Clover and HIV-2 Env WT fused to mRuby2; ( B ) BST-2 fused Clover and HIV-2 Env 1–749 fused mRuby2; ( C ) BST-2 fused Clover and HIV-2 Env N659D fused mRuby2 and ( D ) BST-2 fused Clover and HIV-2 Nef fused mRuby2. Numbers indicated in gate P4 give the percentage of FRET-positive cells included in P2 according to the previous fluorescence-activated cell sorting (FACS) configuration for the different co-expression in living cells ( Figure 6 ); ( E ) Bar diagram summarizing the FRET-positive signals from four independent experiments ( n = 4). Statistical tests give a significance with a p

    Article Snippet: After several passages, infected cells were sorted using a BD FACSAria™ III Cell Sorter (BD Biosciences, San Jose, CA, USA) according to GFP emission since Cas9 protein was fused to GFP.

    Techniques: Expressing, Fluorescence, FACS

    BD FACSAria™ III cell sorter configuration and experimental setup of Förster resonance energy transfer (FRET)-measurements. Firstly, gate P1 selecting living cells (according to forward and sideward scatter FSC-A/SSC-A) and gate P2 selecting cells in P1 (according to forward and sideward scatter FSC-W/SSC-W in order to exclude joined or grouped cells), have been applied (not shown in this figure). Secondly, HEK293T cells expressing Clover or mRuby2 individually, in combination or as a fusion protein, were analyzed with three different filters in order to construct analysis gates and to define the FRET-positive signal. Clover (green fluorescent protein variant) was excited with the laser 488 nm and the FITC filter was used to examine fluorescence emission. mRuby2 (red fluorescent protein variant) was excited with the laser 561 nm and the PE-mCherry filter was used (panel A ). Importantly, mRuby2 and Clover showed some emission in the FRET-channel (PerCP-Cy 5.5 filter). Thus, a gate (P3) was constructed to exclude cells that emitted a false-positive signal in the FRET-channel (panel B ). As described in this image, cells co-transfected with Clover and mRuby2 exerted an aleatory FRET signal that should also be excluded. Therefore, an analysis gate (P4, in red) was applied to determine the FRET-positive cells when the FRET-adapted filters were selected (PerCP-Cy 5.5 and FITC filters, panel C ). This P4 gate excluded cells that were co-transfected with Clover and mRuby2 and thus are FRET-negative (0.5% of P2), while including cells that showed a FRET-positive signal (Clover fused mRuby2; 51.8% of P2). This gating strategy allowed for assessment the enhanced emission of the acceptor fluorochrome and therefore demonstrated the energy transfer between the two fluorochromes; namely the protein interactions.

    Journal: Viruses

    Article Title: Single Amino Acid Substitution N659D in HIV-2 Envelope Glycoprotein (Env) Impairs Viral Release and Hampers BST-2 Antagonism

    doi: 10.3390/v8100285

    Figure Lengend Snippet: BD FACSAria™ III cell sorter configuration and experimental setup of Förster resonance energy transfer (FRET)-measurements. Firstly, gate P1 selecting living cells (according to forward and sideward scatter FSC-A/SSC-A) and gate P2 selecting cells in P1 (according to forward and sideward scatter FSC-W/SSC-W in order to exclude joined or grouped cells), have been applied (not shown in this figure). Secondly, HEK293T cells expressing Clover or mRuby2 individually, in combination or as a fusion protein, were analyzed with three different filters in order to construct analysis gates and to define the FRET-positive signal. Clover (green fluorescent protein variant) was excited with the laser 488 nm and the FITC filter was used to examine fluorescence emission. mRuby2 (red fluorescent protein variant) was excited with the laser 561 nm and the PE-mCherry filter was used (panel A ). Importantly, mRuby2 and Clover showed some emission in the FRET-channel (PerCP-Cy 5.5 filter). Thus, a gate (P3) was constructed to exclude cells that emitted a false-positive signal in the FRET-channel (panel B ). As described in this image, cells co-transfected with Clover and mRuby2 exerted an aleatory FRET signal that should also be excluded. Therefore, an analysis gate (P4, in red) was applied to determine the FRET-positive cells when the FRET-adapted filters were selected (PerCP-Cy 5.5 and FITC filters, panel C ). This P4 gate excluded cells that were co-transfected with Clover and mRuby2 and thus are FRET-negative (0.5% of P2), while including cells that showed a FRET-positive signal (Clover fused mRuby2; 51.8% of P2). This gating strategy allowed for assessment the enhanced emission of the acceptor fluorochrome and therefore demonstrated the energy transfer between the two fluorochromes; namely the protein interactions.

    Article Snippet: After several passages, infected cells were sorted using a BD FACSAria™ III Cell Sorter (BD Biosciences, San Jose, CA, USA) according to GFP emission since Cas9 protein was fused to GFP.

    Techniques: Förster Resonance Energy Transfer, Expressing, Construct, Variant Assay, Fluorescence, Transfection

    RNA-seq of stained and sorted cell populations. A : Bioanalyzer profiles of total RNA derived from unprocessed cells (harvested directly from trypsinised cells) compared to cells that have undergone glyoxal fixation, permeabilisation with 100% methanol and indirect immunofluorescence for CCNB1. 0.1–5 ng of total RNA was separated on total RNA pico chips on Agilent 2100 Bioanalyzer. B : Scatter plots comparing the normalised read count for each annotated gene in GRCh38 in poly(A)+ RNA-seq libraries derived from unprocessed and from glyoxal fixed, permeabilised and stained cells. Two biological replicates were sequenced for each condition and averaged, read counts were normalised to the total number of reads in each library. Data is shown for both COLO205 and MCF-7 cells. C : Flow cytometry density plots for MCF-7 cells labelled with anti-CCNB1 primary antibody and donkey Alexa Fluor-488 conjugated secondary antibody and sorted using a BD FACSAria III sorter. Intact cells (shown within the elliptical gate) were distinguished from off-scale events and cell debris using forward scatter (FSC) and side scatter (SSC) measurements (panel 1). Doublets were excluded from the gated cells by a 2-step gating strategy with pulse height (H) plotted against area (A) for FSC parameter followed by SSC-H versus SSC-A plot (panel 3). Fluorescence thresholds for isolation of CCNB1 positive and negative cell fractions are shown in panel 4. Gates were set using the negative control and the CCNB1 positive and negative sorting gates were moved apart from each other to maximise purity when sorted. D : Fluorescence histogram of DAPI intensities for sorted CCNB1 negative and positive populations. E : Scatter plots comparing the normalised read count for each annotated gene in GRCh38 in poly(A)+ RNA-seq libraries derived from two biological replicates each of CCNB1 positive (left) and negative (right) MCF-7 cells. Note the particularly tight correlation after sorting towards the antigen of interest. E : Hierarchical clustering analysis of the 161 significantly different expressed genes identified DEseq2 analysis (p

    Journal: PLoS ONE

    Article Title: Glyoxal fixation facilitates transcriptome analysis after antigen staining and cell sorting by flow cytometry

    doi: 10.1371/journal.pone.0240769

    Figure Lengend Snippet: RNA-seq of stained and sorted cell populations. A : Bioanalyzer profiles of total RNA derived from unprocessed cells (harvested directly from trypsinised cells) compared to cells that have undergone glyoxal fixation, permeabilisation with 100% methanol and indirect immunofluorescence for CCNB1. 0.1–5 ng of total RNA was separated on total RNA pico chips on Agilent 2100 Bioanalyzer. B : Scatter plots comparing the normalised read count for each annotated gene in GRCh38 in poly(A)+ RNA-seq libraries derived from unprocessed and from glyoxal fixed, permeabilised and stained cells. Two biological replicates were sequenced for each condition and averaged, read counts were normalised to the total number of reads in each library. Data is shown for both COLO205 and MCF-7 cells. C : Flow cytometry density plots for MCF-7 cells labelled with anti-CCNB1 primary antibody and donkey Alexa Fluor-488 conjugated secondary antibody and sorted using a BD FACSAria III sorter. Intact cells (shown within the elliptical gate) were distinguished from off-scale events and cell debris using forward scatter (FSC) and side scatter (SSC) measurements (panel 1). Doublets were excluded from the gated cells by a 2-step gating strategy with pulse height (H) plotted against area (A) for FSC parameter followed by SSC-H versus SSC-A plot (panel 3). Fluorescence thresholds for isolation of CCNB1 positive and negative cell fractions are shown in panel 4. Gates were set using the negative control and the CCNB1 positive and negative sorting gates were moved apart from each other to maximise purity when sorted. D : Fluorescence histogram of DAPI intensities for sorted CCNB1 negative and positive populations. E : Scatter plots comparing the normalised read count for each annotated gene in GRCh38 in poly(A)+ RNA-seq libraries derived from two biological replicates each of CCNB1 positive (left) and negative (right) MCF-7 cells. Note the particularly tight correlation after sorting towards the antigen of interest. E : Hierarchical clustering analysis of the 161 significantly different expressed genes identified DEseq2 analysis (p

    Article Snippet: Cells were sorted using a 100 μm nozzle (at 20PSI) on a BD FACSARIA III cell sorter (BD Biosciences, UK) with cooling of the sample and collection chambers enabled.

    Techniques: RNA Sequencing Assay, Staining, Derivative Assay, Immunofluorescence, Flow Cytometry, Fluorescence, Isolation, Negative Control