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Becton Dickinson bd facs aria ii
Fluorescence analysis of 2-NBDG uptake by flow cytometry. <t>FACS</t> analysis of 2-NBDG uptake in differentiated L6 cells by plotting cell count against <t>FITC</t> revealed that 8%, 8.1% and 30% of cells uptake 2-NBDG in control, TBHP and Rosiglitazone treated cells respectively whereas 30.6%, 33.1%, 28%, 32% of cells uptake 2-NBDG, pretreated with two different concentrations (10 and 100 μM) of Naringin along with/without TBHP respectively. Each value represents mean ± SD (standard deviation) from triplicate measurements (n = 3) of three different experiments. Significance test between various groups was determined by using one way ANOVA followed by Duncan’s multiple range test. * P≤0.05 versus control.
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1) Product Images from "Preconditioning L6 Muscle Cells with Naringin Ameliorates Oxidative Stress and Increases Glucose Uptake"

Article Title: Preconditioning L6 Muscle Cells with Naringin Ameliorates Oxidative Stress and Increases Glucose Uptake

Journal: PLoS ONE

doi: 10.1371/journal.pone.0132429

Fluorescence analysis of 2-NBDG uptake by flow cytometry. FACS analysis of 2-NBDG uptake in differentiated L6 cells by plotting cell count against FITC revealed that 8%, 8.1% and 30% of cells uptake 2-NBDG in control, TBHP and Rosiglitazone treated cells respectively whereas 30.6%, 33.1%, 28%, 32% of cells uptake 2-NBDG, pretreated with two different concentrations (10 and 100 μM) of Naringin along with/without TBHP respectively. Each value represents mean ± SD (standard deviation) from triplicate measurements (n = 3) of three different experiments. Significance test between various groups was determined by using one way ANOVA followed by Duncan’s multiple range test. * P≤0.05 versus control.
Figure Legend Snippet: Fluorescence analysis of 2-NBDG uptake by flow cytometry. FACS analysis of 2-NBDG uptake in differentiated L6 cells by plotting cell count against FITC revealed that 8%, 8.1% and 30% of cells uptake 2-NBDG in control, TBHP and Rosiglitazone treated cells respectively whereas 30.6%, 33.1%, 28%, 32% of cells uptake 2-NBDG, pretreated with two different concentrations (10 and 100 μM) of Naringin along with/without TBHP respectively. Each value represents mean ± SD (standard deviation) from triplicate measurements (n = 3) of three different experiments. Significance test between various groups was determined by using one way ANOVA followed by Duncan’s multiple range test. * P≤0.05 versus control.

Techniques Used: Fluorescence, Flow Cytometry, Cytometry, FACS, Cell Counting, Standard Deviation

2) Product Images from "Coral cell separation and isolation by fluorescence-activated cell sorting (FACS)"

Article Title: Coral cell separation and isolation by fluorescence-activated cell sorting (FACS)

Journal: BMC Cell Biology

doi: 10.1186/s12860-017-0146-8

Coral cell differentiation patterns. P. damicornis cell suspension was incubated with different fluorescent markers and analyzed on FACS Accuri C6. All of the samples were analyzed on two dimensional plots of the green channel- 488 nm (533/30BP) and on the far-red channel-640 nm (675/25BP) to differentiate the Symbiodinium positive cells. Gated in black are the differentiated populations with their cell percentage indicated. The analysis was done after differentiation of cells and debris on FSC and SSC and live/dead cells using PI. a ) Background control of unstained cells for all of the panels. In the gate are the Symbiodinium cells positive at 640 nm (675/25BP) due to natural fluorescence. b ) Labeled cells with Phen Green, a color that is quenched by bi-valent metals. A small population of cells had the lowest level of metal and it differentiated from the main population. c ) Labeled cells with ALDEFLUOR that indicates high levels of ALDH activity. No clear differentiation was found, but the width of the signal in different cells is 3 fluorescent log scales which enables us to differentiate low, moderate and high labeling. d ) Cells containing ROS, labeled with CellRox. Clear negative and positive populations (positive marked) were identified. Note that all the Symbiodinium positive cells are positive to ROS. e ) Cells were incubated with glucose analog 2-NBDG. Positive labeling was found, but no clear differentiation. f ) Cells were co-incubated for 3 h with green fluorescent beads for analysis of engulfment. Cells positive to beads are marked
Figure Legend Snippet: Coral cell differentiation patterns. P. damicornis cell suspension was incubated with different fluorescent markers and analyzed on FACS Accuri C6. All of the samples were analyzed on two dimensional plots of the green channel- 488 nm (533/30BP) and on the far-red channel-640 nm (675/25BP) to differentiate the Symbiodinium positive cells. Gated in black are the differentiated populations with their cell percentage indicated. The analysis was done after differentiation of cells and debris on FSC and SSC and live/dead cells using PI. a ) Background control of unstained cells for all of the panels. In the gate are the Symbiodinium cells positive at 640 nm (675/25BP) due to natural fluorescence. b ) Labeled cells with Phen Green, a color that is quenched by bi-valent metals. A small population of cells had the lowest level of metal and it differentiated from the main population. c ) Labeled cells with ALDEFLUOR that indicates high levels of ALDH activity. No clear differentiation was found, but the width of the signal in different cells is 3 fluorescent log scales which enables us to differentiate low, moderate and high labeling. d ) Cells containing ROS, labeled with CellRox. Clear negative and positive populations (positive marked) were identified. Note that all the Symbiodinium positive cells are positive to ROS. e ) Cells were incubated with glucose analog 2-NBDG. Positive labeling was found, but no clear differentiation. f ) Cells were co-incubated for 3 h with green fluorescent beads for analysis of engulfment. Cells positive to beads are marked

Techniques Used: Cell Differentiation, Incubation, FACS, Fluorescence, Labeling, Activity Assay

Examples of cell differentiation from a chemical compound screening. The P. damicornis cell suspension was incubated with the indicated chemical compounds for 30 min at 20 o C and analyzed by FACS LSRFortessa. The analysis was done after differentiation of cells and debris on FSC and SSC and live/dead cells using DAPI, and separated to Symbiodinium positive cells ( e – h ) and Symbiodinium negative cells ( a – d , as shown in Fig. 1 ). a ) Background control of unstained Symbiodinium negative cells for ( b – d) . b ) Labeled cells with CAY10455, had separation for 3 potential populations shown in gates. c ) Labeled cells with Rhodamine Phenylglyoxal had separation of 4 potential populations. d ) Labeled cells with Acridine Orange had separation of 3 populations shown in red. e ) Background control of unstained Symbiodinium positive cells for ( f) . f ) Labeled cells with Dihydroethidium had separation of 3 populations positive to Symbiodinium . g ) Background control of unstained Symbiodinium positive cells for ( h ). h ) Labeled cells with Acridine Orange had separation of 3 populations positive to Symbiodinium
Figure Legend Snippet: Examples of cell differentiation from a chemical compound screening. The P. damicornis cell suspension was incubated with the indicated chemical compounds for 30 min at 20 o C and analyzed by FACS LSRFortessa. The analysis was done after differentiation of cells and debris on FSC and SSC and live/dead cells using DAPI, and separated to Symbiodinium positive cells ( e – h ) and Symbiodinium negative cells ( a – d , as shown in Fig. 1 ). a ) Background control of unstained Symbiodinium negative cells for ( b – d) . b ) Labeled cells with CAY10455, had separation for 3 potential populations shown in gates. c ) Labeled cells with Rhodamine Phenylglyoxal had separation of 4 potential populations. d ) Labeled cells with Acridine Orange had separation of 3 populations shown in red. e ) Background control of unstained Symbiodinium positive cells for ( f) . f ) Labeled cells with Dihydroethidium had separation of 3 populations positive to Symbiodinium . g ) Background control of unstained Symbiodinium positive cells for ( h ). h ) Labeled cells with Acridine Orange had separation of 3 populations positive to Symbiodinium

Techniques Used: Cell Differentiation, Incubation, FACS, Labeling

3) Product Images from "Naive human B cells can neutralize SARS-CoV-2 through recognition of its receptor binding domain"

Article Title: Naive human B cells can neutralize SARS-CoV-2 through recognition of its receptor binding domain

Journal: bioRxiv

doi: 10.1101/2021.02.02.429458

PBMC flow cytometry analyses. ( A ) Representative gating strategy used for FACS of PBMCs pooled from donors 1 and 2. Gating was on naive B cells defined by single living lymphocytes that were CD19 + CD3 - IgD + IgG - . Sorted cells were RBM-specific as defined by spike-PE + /spike-APC + /RBD-Fc-BV650 + /ΔRBM-Fc-BC650-. Sort gate is denoted by the blue arrow. The bottom right plot shows CD27 staining of sorted RBM-specific naive B cells. ( B ) Flow cytometry showing the sort gate and percentage of RBM-specific B cells for the remaining 6 healthy human donors. ( C ) RBM-specific B cell frequency among CD27 + and CD27 - cells. Each symbol represents a different donor ( n = 8).
Figure Legend Snippet: PBMC flow cytometry analyses. ( A ) Representative gating strategy used for FACS of PBMCs pooled from donors 1 and 2. Gating was on naive B cells defined by single living lymphocytes that were CD19 + CD3 - IgD + IgG - . Sorted cells were RBM-specific as defined by spike-PE + /spike-APC + /RBD-Fc-BV650 + /ΔRBM-Fc-BC650-. Sort gate is denoted by the blue arrow. The bottom right plot shows CD27 staining of sorted RBM-specific naive B cells. ( B ) Flow cytometry showing the sort gate and percentage of RBM-specific B cells for the remaining 6 healthy human donors. ( C ) RBM-specific B cell frequency among CD27 + and CD27 - cells. Each symbol represents a different donor ( n = 8).

Techniques Used: Flow Cytometry, FACS, Staining

4) Product Images from "Bioengineered constructs combined with exercise enhance stem cell-mediated treatment of volumetric muscle loss"

Article Title: Bioengineered constructs combined with exercise enhance stem cell-mediated treatment of volumetric muscle loss

Journal: Nature Communications

doi: 10.1038/ncomms15613

MRCs support MuSCs in de novo myofibre formation. ( a ) Representative FACS plot of MRCs. Lower limb muscles were dissected and digested to obtain a mononucleated cellular suspension. These cells were marked using cell specific surface antigens as described in the ‘Results’ and in the ‘Methods’ sections and were analysed using FACS. Five populations were isolated: muscle stem cells (MuSCs), hematopoietic cells (HCs), endothelial cells (ECs), fibro-adipogenic progenitor cells (FAPs) and fibroblast-like cells (FLCs). The relative percentages of each cell population are 10%, 25%, 39%, 8% and 18%, respectively ( n =6). ( b ) Quantified results of in vitro bioluminescence generated from cultured bioconstructs containing Luc + MuSCs, either alone (MuSC + /MRC − ) or in combination with Luc − MRCs (MuSC + /MRC + ). Bioconstructs were cultured for three days, and bioluminescence was measured each day ( n =4). ( c ) Representative images of bioluminescence measured from mice 10 days after transplantation of bioconstructs in left TA muscles immediately following VML injury ( d ) Quantified results of non-invasive imaging of transplanted bioconstructs. Bioconstructs with no cells (MuSC − /MRC − ), Luc + MuSCs (MuSC + /MRC − ), or Luc + MuSCs in addition to Luc − MRCs (MuSC + /MRC + ) were transplanted into TA muscles that had received VML injuries. Bioluminescence was measured 10 days following transplantation ( n =4). Data are±s.e.m. For statistical analysis, t -tests were used. * P
Figure Legend Snippet: MRCs support MuSCs in de novo myofibre formation. ( a ) Representative FACS plot of MRCs. Lower limb muscles were dissected and digested to obtain a mononucleated cellular suspension. These cells were marked using cell specific surface antigens as described in the ‘Results’ and in the ‘Methods’ sections and were analysed using FACS. Five populations were isolated: muscle stem cells (MuSCs), hematopoietic cells (HCs), endothelial cells (ECs), fibro-adipogenic progenitor cells (FAPs) and fibroblast-like cells (FLCs). The relative percentages of each cell population are 10%, 25%, 39%, 8% and 18%, respectively ( n =6). ( b ) Quantified results of in vitro bioluminescence generated from cultured bioconstructs containing Luc + MuSCs, either alone (MuSC + /MRC − ) or in combination with Luc − MRCs (MuSC + /MRC + ). Bioconstructs were cultured for three days, and bioluminescence was measured each day ( n =4). ( c ) Representative images of bioluminescence measured from mice 10 days after transplantation of bioconstructs in left TA muscles immediately following VML injury ( d ) Quantified results of non-invasive imaging of transplanted bioconstructs. Bioconstructs with no cells (MuSC − /MRC − ), Luc + MuSCs (MuSC + /MRC − ), or Luc + MuSCs in addition to Luc − MRCs (MuSC + /MRC + ) were transplanted into TA muscles that had received VML injuries. Bioluminescence was measured 10 days following transplantation ( n =4). Data are±s.e.m. For statistical analysis, t -tests were used. * P

Techniques Used: FACS, Isolation, In Vitro, Generated, Cell Culture, Mouse Assay, Transplantation Assay, Imaging

5) Product Images from "A novel immunogenic mouse model of melanoma for the preclinical assessment of combination targeted and immune-based therapy"

Article Title: A novel immunogenic mouse model of melanoma for the preclinical assessment of combination targeted and immune-based therapy

Journal: Scientific Reports

doi: 10.1038/s41598-018-37883-y

Expression of the immunogen, ovalbumin, in YUMM1.1 tumor cells promotes T cell-mediated tumor control. ( a ) Tumor growth and survival of 3 × 10 5 YUMM1.1 cells in C57BL/6 mice or NSG mice, with survival measured as time for tumors to reach > 1200 mm 3 . ns – not significant, log-rank (Mantel-Cox) test, n = 5–8. ( b ) YUMM1.1-OVA sorted by FACS into low and high GFP-expressing populations; YOVAL1.1 and YOVAH1.1, respectively. ( c ) Killing by OT-I T cells co-cultured for 4 hours at indicated ratios with 51Cr-labelled target cells pre-stimulated +/− IFNγ. One way ANOVA, Tukey’s multiple comparisons test, n = 3. ( d ) YOVAL1.1 tumor growth and survival in C57BL/6 mice or NSG mice with survival measured as time for tumors to reach > 1200 mm 3 , log-rank (Mantel-Cox) test, n = 3–5. ( e ) Growth of YOVAL1.1 in NSG or Rag1 −/− mice, n = 3. ( f ) YOVAL1.1 tumor growth and survival following transfer of activated OT-I T cells or PBS. YV1.1 – YUMM1.1 transduced with empty vector. All error bars show ±SEM. **p
Figure Legend Snippet: Expression of the immunogen, ovalbumin, in YUMM1.1 tumor cells promotes T cell-mediated tumor control. ( a ) Tumor growth and survival of 3 × 10 5 YUMM1.1 cells in C57BL/6 mice or NSG mice, with survival measured as time for tumors to reach > 1200 mm 3 . ns – not significant, log-rank (Mantel-Cox) test, n = 5–8. ( b ) YUMM1.1-OVA sorted by FACS into low and high GFP-expressing populations; YOVAL1.1 and YOVAH1.1, respectively. ( c ) Killing by OT-I T cells co-cultured for 4 hours at indicated ratios with 51Cr-labelled target cells pre-stimulated +/− IFNγ. One way ANOVA, Tukey’s multiple comparisons test, n = 3. ( d ) YOVAL1.1 tumor growth and survival in C57BL/6 mice or NSG mice with survival measured as time for tumors to reach > 1200 mm 3 , log-rank (Mantel-Cox) test, n = 3–5. ( e ) Growth of YOVAL1.1 in NSG or Rag1 −/− mice, n = 3. ( f ) YOVAL1.1 tumor growth and survival following transfer of activated OT-I T cells or PBS. YV1.1 – YUMM1.1 transduced with empty vector. All error bars show ±SEM. **p

Techniques Used: Expressing, Mouse Assay, FACS, Cell Culture, Transduction, Plasmid Preparation

6) Product Images from "Transcriptional Circuit Fragility Influences HIV Proviral Fate"

Article Title: Transcriptional Circuit Fragility Influences HIV Proviral Fate

Journal: bioRxiv

doi: 10.1101/504969

The Master Regulator of the Viral Phase (Tat) Operates in a KAP1-independent Manner (A) Quantification of firefly luciferase activity (FFL) from an HIV LTR-FFL reporter in the absence (0) or presence of increasing Tat concentrations (normalized to a constitutive CMV-Renilla (RL)) in the shNT and shKAP1 U2OS cell lines. (B) Western blots of the indicated U2OS cell lines used in panel (A). (C) Experimental outline in which the HIV Tat-shNT and shKAP1 cell lines were transduced with pTRIP-LUC or pTRIP-Tat lentiviruses at day 1 and collected at day 3 for their analysis by FACS and RT-qPCR assays (panels (D) and (E), respectively). (D) Flow cytometry based quantitation of the percentage of GFP+ cells after transduction of the shNT and shKAP1 cell lines from panel (C) with the indicated lentiviruses expressing LUC and Tat. (E) RT-qPCR assay of RNA isolated from the cell lines from panel (C) using the elongation amplicon (+2627). The change in HIV gene expression is shown as fold HIV RNA activation (Tat over LUC). (F) Top, scheme of the HIV Tat-(2B2D) provirus. The arrow denotes the position of the transcription start site (TSS) on the 5’-LTR. Bottom, ChIP assays were performed with protein extracts from the four cells states from panel (C) and the indicated antibodies (KAP1, CDK9, and Pol II) followed by qPCR with a series of amplicons mapping throughout the entire provirus (indicated at the top of the schematic) to monitor factor interactions with the HIV genome. A normal IgG serum was used as a negative control in ChIP-qPCR, which revealed no amplification (not shown). The ChIP-qPCR data was normalized using the “Percent Input Method” (see Experimental Procedures for full description of normalization method). Values represent the percentage (%) of Input DNA immunoprecipitated (IP DNA) and are the average of three independent experiments (mean ± SEM; n =3).
Figure Legend Snippet: The Master Regulator of the Viral Phase (Tat) Operates in a KAP1-independent Manner (A) Quantification of firefly luciferase activity (FFL) from an HIV LTR-FFL reporter in the absence (0) or presence of increasing Tat concentrations (normalized to a constitutive CMV-Renilla (RL)) in the shNT and shKAP1 U2OS cell lines. (B) Western blots of the indicated U2OS cell lines used in panel (A). (C) Experimental outline in which the HIV Tat-shNT and shKAP1 cell lines were transduced with pTRIP-LUC or pTRIP-Tat lentiviruses at day 1 and collected at day 3 for their analysis by FACS and RT-qPCR assays (panels (D) and (E), respectively). (D) Flow cytometry based quantitation of the percentage of GFP+ cells after transduction of the shNT and shKAP1 cell lines from panel (C) with the indicated lentiviruses expressing LUC and Tat. (E) RT-qPCR assay of RNA isolated from the cell lines from panel (C) using the elongation amplicon (+2627). The change in HIV gene expression is shown as fold HIV RNA activation (Tat over LUC). (F) Top, scheme of the HIV Tat-(2B2D) provirus. The arrow denotes the position of the transcription start site (TSS) on the 5’-LTR. Bottom, ChIP assays were performed with protein extracts from the four cells states from panel (C) and the indicated antibodies (KAP1, CDK9, and Pol II) followed by qPCR with a series of amplicons mapping throughout the entire provirus (indicated at the top of the schematic) to monitor factor interactions with the HIV genome. A normal IgG serum was used as a negative control in ChIP-qPCR, which revealed no amplification (not shown). The ChIP-qPCR data was normalized using the “Percent Input Method” (see Experimental Procedures for full description of normalization method). Values represent the percentage (%) of Input DNA immunoprecipitated (IP DNA) and are the average of three independent experiments (mean ± SEM; n =3).

Techniques Used: Luciferase, Activity Assay, Western Blot, Transduction, FACS, Quantitative RT-PCR, Flow Cytometry, Quantitation Assay, Expressing, Isolation, Amplification, Activation Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Negative Control, Immunoprecipitation

7) Product Images from "A novel immunogenic mouse model of melanoma for the preclinical assessment of combination targeted and immune-based therapy"

Article Title: A novel immunogenic mouse model of melanoma for the preclinical assessment of combination targeted and immune-based therapy

Journal: Scientific Reports

doi: 10.1038/s41598-018-37883-y

Expression of the immunogen, ovalbumin, in YUMM1.1 tumor cells promotes T cell-mediated tumor control. ( a ) Tumor growth and survival of 3 × 10 5 YUMM1.1 cells in C57BL/6 mice or NSG mice, with survival measured as time for tumors to reach > 1200 mm 3 . ns – not significant, log-rank (Mantel-Cox) test, n = 5–8. ( b ) YUMM1.1-OVA sorted by FACS into low and high GFP-expressing populations; YOVAL1.1 and YOVAH1.1, respectively. ( c ) Killing by OT-I T cells co-cultured for 4 hours at indicated ratios with 51Cr-labelled target cells pre-stimulated +/− IFNγ. One way ANOVA, Tukey’s multiple comparisons test, n = 3. ( d ) YOVAL1.1 tumor growth and survival in C57BL/6 mice or NSG mice with survival measured as time for tumors to reach > 1200 mm 3 , log-rank (Mantel-Cox) test, n = 3–5. ( e ) Growth of YOVAL1.1 in NSG or Rag1 −/− mice, n = 3. ( f ) YOVAL1.1 tumor growth and survival following transfer of activated OT-I T cells or PBS. YV1.1 – YUMM1.1 transduced with empty vector. All error bars show ±SEM. **p
Figure Legend Snippet: Expression of the immunogen, ovalbumin, in YUMM1.1 tumor cells promotes T cell-mediated tumor control. ( a ) Tumor growth and survival of 3 × 10 5 YUMM1.1 cells in C57BL/6 mice or NSG mice, with survival measured as time for tumors to reach > 1200 mm 3 . ns – not significant, log-rank (Mantel-Cox) test, n = 5–8. ( b ) YUMM1.1-OVA sorted by FACS into low and high GFP-expressing populations; YOVAL1.1 and YOVAH1.1, respectively. ( c ) Killing by OT-I T cells co-cultured for 4 hours at indicated ratios with 51Cr-labelled target cells pre-stimulated +/− IFNγ. One way ANOVA, Tukey’s multiple comparisons test, n = 3. ( d ) YOVAL1.1 tumor growth and survival in C57BL/6 mice or NSG mice with survival measured as time for tumors to reach > 1200 mm 3 , log-rank (Mantel-Cox) test, n = 3–5. ( e ) Growth of YOVAL1.1 in NSG or Rag1 −/− mice, n = 3. ( f ) YOVAL1.1 tumor growth and survival following transfer of activated OT-I T cells or PBS. YV1.1 – YUMM1.1 transduced with empty vector. All error bars show ±SEM. **p

Techniques Used: Expressing, Mouse Assay, FACS, Cell Culture, Transduction, Plasmid Preparation

8) Product Images from "An integrated multi-omic single cell atlas to redefine human B cell memory"

Article Title: An integrated multi-omic single cell atlas to redefine human B cell memory

Journal: bioRxiv

doi: 10.1101/801530

CD45RB marks human memory B cells and identifies a unique early memory population A) Computational workflow for UMAP generation. UMAP coordinates were generated using conserved molecules as input on a random subset of donor-pooled cells from each of the twelve mass cytometry panels of the screen. B) UMAP plots colored by individually-scaled expression of the indicated molecule. Molecules are organized and colored by function or by their identity as a conserved molecule (right column). Molecules in each row are highly correlated (Pearson method, ± r > 0.3) with the conserved molecule in their row. The canonical memory marker (CD27) and putative memory marker (CD45RB) are boxed. Arrows indicate CD45RB+ CD27- cells. C) UMAP plot colored by canonical B cell subset. Circles indicate islands of cells with memory-like phenotypes based on UMAP co-localization and high-dimensional expression profiles. Arrows are the same as in B) and indicate cells with memory-like phenotypes that are not identified as memory cells by the canonical gating scheme. D) CD38 lo B cells (non-transitional B cells) are plotted on a biaxial and colored by their canonical B cell gate. Percent of CD38 lo B cells in each quadrant is quantified (red text). E) Percent of cells in each quadrant from D) by canonical B cell subset. High percentage of ungated cells in quadrant II is boxed and bolded. F) Experimental workflow for immune repertoire analysis. Peripheral blood was drawn from healthy, human donors (n=2), density gradient centrifugation was performed, and cell preparations were magnetically depleted of non-B cells. Singlet Viable CD45+ lin- CD19+ CD38 lo B cells were FACS-sorted from the four quadrants of CD27 x CD45RB, gDNA was extracted, and the IgH locus was sequenced by NGS. G) Mutation frequency of the IgH loci outside of the CDR3 region for sorted populations, colored by mutation type. Data are pooled from donors. All pairwise comparisons of populations were significantly different (Wilcoxon rank sum test with Bonferroni correction, p
Figure Legend Snippet: CD45RB marks human memory B cells and identifies a unique early memory population A) Computational workflow for UMAP generation. UMAP coordinates were generated using conserved molecules as input on a random subset of donor-pooled cells from each of the twelve mass cytometry panels of the screen. B) UMAP plots colored by individually-scaled expression of the indicated molecule. Molecules are organized and colored by function or by their identity as a conserved molecule (right column). Molecules in each row are highly correlated (Pearson method, ± r > 0.3) with the conserved molecule in their row. The canonical memory marker (CD27) and putative memory marker (CD45RB) are boxed. Arrows indicate CD45RB+ CD27- cells. C) UMAP plot colored by canonical B cell subset. Circles indicate islands of cells with memory-like phenotypes based on UMAP co-localization and high-dimensional expression profiles. Arrows are the same as in B) and indicate cells with memory-like phenotypes that are not identified as memory cells by the canonical gating scheme. D) CD38 lo B cells (non-transitional B cells) are plotted on a biaxial and colored by their canonical B cell gate. Percent of CD38 lo B cells in each quadrant is quantified (red text). E) Percent of cells in each quadrant from D) by canonical B cell subset. High percentage of ungated cells in quadrant II is boxed and bolded. F) Experimental workflow for immune repertoire analysis. Peripheral blood was drawn from healthy, human donors (n=2), density gradient centrifugation was performed, and cell preparations were magnetically depleted of non-B cells. Singlet Viable CD45+ lin- CD19+ CD38 lo B cells were FACS-sorted from the four quadrants of CD27 x CD45RB, gDNA was extracted, and the IgH locus was sequenced by NGS. G) Mutation frequency of the IgH loci outside of the CDR3 region for sorted populations, colored by mutation type. Data are pooled from donors. All pairwise comparisons of populations were significantly different (Wilcoxon rank sum test with Bonferroni correction, p

Techniques Used: Generated, Mass Cytometry, Expressing, Marker, Gradient Centrifugation, FACS, Next-Generation Sequencing, Mutagenesis

9) Product Images from "Quantification of phenolics in Syzygium cumini seed and their modulatory role on tertiary butyl-hydrogen peroxide-induced oxidative stress in H9c2 cell lines and key enzymes in cardioprotection"

Article Title: Quantification of phenolics in Syzygium cumini seed and their modulatory role on tertiary butyl-hydrogen peroxide-induced oxidative stress in H9c2 cell lines and key enzymes in cardioprotection

Journal: Journal of Food Science and Technology

doi: 10.1007/s13197-017-2651-3

Fluorescent analysis of ROS by flow cytometry. FACS analysis of intracellular ROS production in H9c2 cell lines by plotting cell count against FITC. The groups contained a control cells, b cells treated with Tertiary butyl hydrogen peroxide (TBHP), c ethyl acetate fractions (100 µg), d methanol fractions (100 µg), e 70% methanol fractions (100 µg), f water fractions (100 µg)
Figure Legend Snippet: Fluorescent analysis of ROS by flow cytometry. FACS analysis of intracellular ROS production in H9c2 cell lines by plotting cell count against FITC. The groups contained a control cells, b cells treated with Tertiary butyl hydrogen peroxide (TBHP), c ethyl acetate fractions (100 µg), d methanol fractions (100 µg), e 70% methanol fractions (100 µg), f water fractions (100 µg)

Techniques Used: Flow Cytometry, Cytometry, FACS, Cell Counting

10) Product Images from "ICER is requisite for Th17 differentiation"

Article Title: ICER is requisite for Th17 differentiation

Journal: Nature Communications

doi: 10.1038/ncomms12993

ICER is expressed in IL-17-producing murine T cells. ( a , left) CREM and β-actin expression in Th17-polarized ICER/CREM +/+ murine CD4 + T cells was measured by western blotting at the indicated time points. ( a , right) CREM expression on day 3 in Th17-polarized ICER/CREM +/+ murine CD4 + T cells (right, far left), Th17-polarized B6.ICER/CREM −/− mice CD4 + T cells (right, second left), empty plasmid transfected HEK-293T cells (right, third left), ICER-overexpressing plasmid-transfected HEK-293T cells (right, third right) or ICERγ-overexpressing plasmid-transfected HEK-293Tcells (right, second right) was measured by western blotting. Data are representative of four experiments. ( b ) ICER/CREM and β-actin expression on day 3 of Th0-, Th1-, Th17- and Treg-polarized ICER/CREM +/+ mice CD4 + T cells was measured by western blotting. Data are representative of three experiments. ( c ) ICER/CREM and β-actin expression of FACS-sorted GFP + (IL-17A-producing cells) or GFP − (IL-17A-non-producing cells) in Th17-polarized B6. IL-17A GFP ICER/CREM +/+ CD4 + T cells on day 3 were measured by western blotting. Data are representative of three experiments. ( d ) ICER and β-actin expression of FACS-sorted IL-17A-producing cells from 18-week-old MRL/lpr mice or naive CD4 T cells from 18-week-old MRL/Mpj control mice was determined by western blotting. A representative (of three) blot is shown (left) and densitometric readings from three experiments are shown on the right (* P
Figure Legend Snippet: ICER is expressed in IL-17-producing murine T cells. ( a , left) CREM and β-actin expression in Th17-polarized ICER/CREM +/+ murine CD4 + T cells was measured by western blotting at the indicated time points. ( a , right) CREM expression on day 3 in Th17-polarized ICER/CREM +/+ murine CD4 + T cells (right, far left), Th17-polarized B6.ICER/CREM −/− mice CD4 + T cells (right, second left), empty plasmid transfected HEK-293T cells (right, third left), ICER-overexpressing plasmid-transfected HEK-293T cells (right, third right) or ICERγ-overexpressing plasmid-transfected HEK-293Tcells (right, second right) was measured by western blotting. Data are representative of four experiments. ( b ) ICER/CREM and β-actin expression on day 3 of Th0-, Th1-, Th17- and Treg-polarized ICER/CREM +/+ mice CD4 + T cells was measured by western blotting. Data are representative of three experiments. ( c ) ICER/CREM and β-actin expression of FACS-sorted GFP + (IL-17A-producing cells) or GFP − (IL-17A-non-producing cells) in Th17-polarized B6. IL-17A GFP ICER/CREM +/+ CD4 + T cells on day 3 were measured by western blotting. Data are representative of three experiments. ( d ) ICER and β-actin expression of FACS-sorted IL-17A-producing cells from 18-week-old MRL/lpr mice or naive CD4 T cells from 18-week-old MRL/Mpj control mice was determined by western blotting. A representative (of three) blot is shown (left) and densitometric readings from three experiments are shown on the right (* P

Techniques Used: Expressing, Western Blot, Mouse Assay, Plasmid Preparation, Transfection, FACS

Human Th17 cells and SLE T cells express increased amounts of ICER. ( a ) The gating strategy used to define and sort the primary lymphocytes memory subsets: Th1 (CD4 + CD45RA − CXCR3 + ); Th2 (CD4 + CD45RA − CCR6 − CCR4 + CXCR3 − ); Th17 (CD4 + CD45RA − CCR6 + CCR4 + ); and Treg (CD4 + CD25 + CD127 − ). ( b ) FACS-sorted Th1, Th2, Th17 and Treg cells were stimulated with plate-bound anti-CD3 and anti-CD28 antibodies for 12 h. ICER and β-actin expression was determined by western blot (* P
Figure Legend Snippet: Human Th17 cells and SLE T cells express increased amounts of ICER. ( a ) The gating strategy used to define and sort the primary lymphocytes memory subsets: Th1 (CD4 + CD45RA − CXCR3 + ); Th2 (CD4 + CD45RA − CCR6 − CCR4 + CXCR3 − ); Th17 (CD4 + CD45RA − CCR6 + CCR4 + ); and Treg (CD4 + CD25 + CD127 − ). ( b ) FACS-sorted Th1, Th2, Th17 and Treg cells were stimulated with plate-bound anti-CD3 and anti-CD28 antibodies for 12 h. ICER and β-actin expression was determined by western blot (* P

Techniques Used: FACS, Expressing, Western Blot

Related Articles

Fluorescence:

Article Title: CRISPR screens identify genomic ribonucleotides as a source of PARP-trapping lesions
Article Snippet: Subsequently, cells positive for both mKO2-Cdt1 and mAG-Geminin fluorescence were collected by sorting on a BD Biosciences FACS Aria II instrument, expanded and used for further experiments. .. Subsequently, cells positive for both mKO2-Cdt1 and mAG-Geminin fluorescence were collected by sorting on a BD Biosciences FACS Aria II instrument, expanded and used for further experiments. .. Expression of mKO2-Cdt1 and mAG-Geminin was confirmed by immunofluorescence and FACS analysis.

Article Title: CRISPR screens identify genomic ribonucleotides as a source of PARP-trapping lesions
Article Snippet: Samples were run on an 8% SDS-PAGE gel and immunoblotting was performed as described above (see ‘Immunobloting’). .. To analyze PARP1 poly(ADP-ribosylation) in a specific phase of the cell cycle, HeLa FUCCI WT or RNASEH2A-KO were trypsinized, washed once with PBS, collected in tubes with PBS supplemented with 3% FCS and 1 µM ADP-HPD, and sorted based on mKO2-Cdt1 (G1 phase) and mAG-Geminin (S/G2/M phases) fluorescence on a BD Biosciences FACS Aria II instrument. .. 4 x 106 FACS-sorted cells were snap-frozen and lysed as described above.

Article Title: Inter-Laboratory Comparison of Extracellular Vesicle Isolation Based on Ultracentrifugation
Article Snippet: Blots were then developed using WesternBright ECL (541004; Biozym Scientific, Germany) and protein bands were detected using the FusionCapt Advanced Solo 4 (Vilber, Germany). .. Fluorescence-Activated Cell Sorting AnalysisFACS measurement of HCT116-derived EVs and their parental cells was performed with BD FACS Canto II, using BD FACSDiva software (BD Biosciences). .. HCT116-derived EVs were captured on anti-human CD9 beads for flow detection (Exosome-Human CD9 beads; Thermo Fisher Scientific).

FACS:

Article Title: CRISPR screens identify genomic ribonucleotides as a source of PARP-trapping lesions
Article Snippet: Subsequently, cells positive for both mKO2-Cdt1 and mAG-Geminin fluorescence were collected by sorting on a BD Biosciences FACS Aria II instrument, expanded and used for further experiments. .. Subsequently, cells positive for both mKO2-Cdt1 and mAG-Geminin fluorescence were collected by sorting on a BD Biosciences FACS Aria II instrument, expanded and used for further experiments. .. Expression of mKO2-Cdt1 and mAG-Geminin was confirmed by immunofluorescence and FACS analysis.

Article Title: CRISPR screens identify genomic ribonucleotides as a source of PARP-trapping lesions
Article Snippet: Samples were run on an 8% SDS-PAGE gel and immunoblotting was performed as described above (see ‘Immunobloting’). .. To analyze PARP1 poly(ADP-ribosylation) in a specific phase of the cell cycle, HeLa FUCCI WT or RNASEH2A-KO were trypsinized, washed once with PBS, collected in tubes with PBS supplemented with 3% FCS and 1 µM ADP-HPD, and sorted based on mKO2-Cdt1 (G1 phase) and mAG-Geminin (S/G2/M phases) fluorescence on a BD Biosciences FACS Aria II instrument. .. 4 x 106 FACS-sorted cells were snap-frozen and lysed as described above.

Article Title: Optimized Surface Markers for the Prospective Isolation of High-Quality hiPSCs using Flow Cytometry Selection
Article Snippet: Immunostaining of cells was carried out using manufacturers' recommended antibody dilution in staining buffer containing Hanks' Balanced Salt Solution (Invitrogen), 4% fetal bovine serum (Invitrogen) and 10 mM Hepes (Invitrogen). .. Flow cytometry sorting was performed on FACS Aria II (BD, Biosciences), and all primary antibodies used were purchased from BD Biosciences. .. Sorting at clonal cell densities was performed by sorting cells directly into 96-well plates.

Article Title: Intracellular Autofluorescence as a New Biomarker for Cancer Stem Cells in Glioblastoma
Article Snippet: In both conditions, adherent and neurospheres cultures were incubated at 37 °C in a humidified atmosphere containing 5% (v/v ) CO2. .. Flow Cytometry Analysis GBM cells were resuspended in FACS flow buffer (BD Biosciences, San Jose, CA, USA) with DAPI (for exclusion of dead cells; 1:1000) before flow cytometry analysis using FACS Canto II (BD Biosciences). ..

Article Title: Inter-Laboratory Comparison of Extracellular Vesicle Isolation Based on Ultracentrifugation
Article Snippet: Blots were then developed using WesternBright ECL (541004; Biozym Scientific, Germany) and protein bands were detected using the FusionCapt Advanced Solo 4 (Vilber, Germany). .. Fluorescence-Activated Cell Sorting AnalysisFACS measurement of HCT116-derived EVs and their parental cells was performed with BD FACS Canto II, using BD FACSDiva software (BD Biosciences). .. HCT116-derived EVs were captured on anti-human CD9 beads for flow detection (Exosome-Human CD9 beads; Thermo Fisher Scientific).

Article Title: CD74 is a regulator of hematopoietic stem cell maintenance
Article Snippet: All analyses were done using a FACS Canto II flow cytometer (BD Bioscience, USA). .. Sorting of the LSK CD45.1 WT and CD45.2 CD74−/− cells was performed using a FACS Aria II system (BD Bioscience, USA), following enrichment using CD117 (c-Kit) MicroBeads (cat: 130-091-224) on LS MACS Separation Columns (cat: 130-042-401), both obtained from Miltenyi Biotec, UK. .. Generation of chimeric mice For the microenvironment experiment: Lethally irradiated (950 Rad) C57BL/6 (WT) recipient mice were reconstituted with 5*106 WT or CD74−/− BM cells.

Article Title: A distinct metabolic state arises during the emergence of 2‐cell‐like cells
Article Snippet: Fluorescence‐assisted cell sortingCells were washed with room temperature sterile PBS, trypsinized and resuspended in ice‐cold sterile 0.5% BSA PBS solution. .. Sorting was performed using a BD BioSciences FACS Aria II or III. ..

Article Title: Macrophages are exploited from an innate wound healing response to facilitate cancer metastasis
Article Snippet: Dead cells and red blood cells were excluded using 1 µg/ml 7AAD (Sigma-Aldrich) and anti-Ter-119 PerCP-Cy5.5 (Ter-119), respectively. .. Flow cytometry was performed on a BD FACS Canto II (BD Biosciences) or cells were sorted for subsequent analyses using a BD FACSAria (BD Biosciences). .. Data were analysed using FlowJo software (Freestar Inc.).

Flow Cytometry:

Article Title: Optimized Surface Markers for the Prospective Isolation of High-Quality hiPSCs using Flow Cytometry Selection
Article Snippet: Immunostaining of cells was carried out using manufacturers' recommended antibody dilution in staining buffer containing Hanks' Balanced Salt Solution (Invitrogen), 4% fetal bovine serum (Invitrogen) and 10 mM Hepes (Invitrogen). .. Flow cytometry sorting was performed on FACS Aria II (BD, Biosciences), and all primary antibodies used were purchased from BD Biosciences. .. Sorting at clonal cell densities was performed by sorting cells directly into 96-well plates.

Article Title: Intracellular Autofluorescence as a New Biomarker for Cancer Stem Cells in Glioblastoma
Article Snippet: In both conditions, adherent and neurospheres cultures were incubated at 37 °C in a humidified atmosphere containing 5% (v/v ) CO2. .. Flow Cytometry Analysis GBM cells were resuspended in FACS flow buffer (BD Biosciences, San Jose, CA, USA) with DAPI (for exclusion of dead cells; 1:1000) before flow cytometry analysis using FACS Canto II (BD Biosciences). ..

Article Title: Macrophages are exploited from an innate wound healing response to facilitate cancer metastasis
Article Snippet: Dead cells and red blood cells were excluded using 1 µg/ml 7AAD (Sigma-Aldrich) and anti-Ter-119 PerCP-Cy5.5 (Ter-119), respectively. .. Flow cytometry was performed on a BD FACS Canto II (BD Biosciences) or cells were sorted for subsequent analyses using a BD FACSAria (BD Biosciences). .. Data were analysed using FlowJo software (Freestar Inc.).

Software:

Article Title: Inter-Laboratory Comparison of Extracellular Vesicle Isolation Based on Ultracentrifugation
Article Snippet: Blots were then developed using WesternBright ECL (541004; Biozym Scientific, Germany) and protein bands were detected using the FusionCapt Advanced Solo 4 (Vilber, Germany). .. Fluorescence-Activated Cell Sorting AnalysisFACS measurement of HCT116-derived EVs and their parental cells was performed with BD FACS Canto II, using BD FACSDiva software (BD Biosciences). .. HCT116-derived EVs were captured on anti-human CD9 beads for flow detection (Exosome-Human CD9 beads; Thermo Fisher Scientific).

Magnetic Cell Separation:

Article Title: CD74 is a regulator of hematopoietic stem cell maintenance
Article Snippet: All analyses were done using a FACS Canto II flow cytometer (BD Bioscience, USA). .. Sorting of the LSK CD45.1 WT and CD45.2 CD74−/− cells was performed using a FACS Aria II system (BD Bioscience, USA), following enrichment using CD117 (c-Kit) MicroBeads (cat: 130-091-224) on LS MACS Separation Columns (cat: 130-042-401), both obtained from Miltenyi Biotec, UK. .. Generation of chimeric mice For the microenvironment experiment: Lethally irradiated (950 Rad) C57BL/6 (WT) recipient mice were reconstituted with 5*106 WT or CD74−/− BM cells.

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  • 86
    Becton Dickinson facs aria ii
    MASTL depletion increases the radiosensitivity of breast cancer cells. <t>MCF7</t> cells were transfected with either 5 nmol/l control siRNA or MASTL.5 siRNA. Cells were treated 0, 3 or 4 Gy irradiation for 42 h. a , b Cell proliferation was determined by <t>FACS</t> analysis and then analyzed by immunoblotted with the indicated antibodies. c The results of the clonogenic assay. Representative images of the cells treated the indicated conditions (left panel). The number of colonies was measured (right panel). d The sphere forming assay was performed ( > 110 sphere colonies per data point). Scale bar = 100 μm. Representative images of sphere forming assay. e PP2A-Aα/β proteins were immunoprecipitated using an anti-PP2A-Aα/β antibody and analyzed for PP2A activity. The data represent typical results and are presented as the mean ± standard deviation of three independent experiments; ** P
    Facs Aria Ii, 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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    Becton Dickinson bd facs aria ii
    Coral cell differentiation patterns. P. damicornis cell suspension was incubated with different fluorescent markers and analyzed on <t>FACS</t> <t>Accuri</t> C6. All of the samples were analyzed on two dimensional plots of the green channel- 488 nm (533/30BP) and on the far-red channel-640 nm (675/25BP) to differentiate the Symbiodinium positive cells. Gated in black are the differentiated populations with their cell percentage indicated. The analysis was done after differentiation of cells and debris on FSC and SSC and live/dead cells using PI. a ) Background control of unstained cells for all of the panels. In the gate are the Symbiodinium cells positive at 640 nm (675/25BP) due to natural fluorescence. b ) Labeled cells with Phen Green, a color that is quenched by bi-valent metals. A small population of cells had the lowest level of metal and it differentiated from the main population. c ) Labeled cells with ALDEFLUOR that indicates high levels of ALDH activity. No clear differentiation was found, but the width of the signal in different cells is 3 fluorescent log scales which enables us to differentiate low, moderate and high labeling. d ) Cells containing ROS, labeled with CellRox. Clear negative and positive populations (positive marked) were identified. Note that all the Symbiodinium positive cells are positive to ROS. e ) Cells were incubated with glucose analog 2-NBDG. Positive labeling was found, but no clear differentiation. f ) Cells were co-incubated for 3 h with green fluorescent beads for analysis of engulfment. Cells positive to beads are marked
    Bd Facs Aria Ii, 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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    MASTL depletion increases the radiosensitivity of breast cancer cells. MCF7 cells were transfected with either 5 nmol/l control siRNA or MASTL.5 siRNA. Cells were treated 0, 3 or 4 Gy irradiation for 42 h. a , b Cell proliferation was determined by FACS analysis and then analyzed by immunoblotted with the indicated antibodies. c The results of the clonogenic assay. Representative images of the cells treated the indicated conditions (left panel). The number of colonies was measured (right panel). d The sphere forming assay was performed ( > 110 sphere colonies per data point). Scale bar = 100 μm. Representative images of sphere forming assay. e PP2A-Aα/β proteins were immunoprecipitated using an anti-PP2A-Aα/β antibody and analyzed for PP2A activity. The data represent typical results and are presented as the mean ± standard deviation of three independent experiments; ** P

    Journal: BMC Cancer

    Article Title: MASTL inhibition promotes mitotic catastrophe through PP2A activation to inhibit cancer growth and radioresistance in breast cancer cells

    doi: 10.1186/s12885-018-4600-6

    Figure Lengend Snippet: MASTL depletion increases the radiosensitivity of breast cancer cells. MCF7 cells were transfected with either 5 nmol/l control siRNA or MASTL.5 siRNA. Cells were treated 0, 3 or 4 Gy irradiation for 42 h. a , b Cell proliferation was determined by FACS analysis and then analyzed by immunoblotted with the indicated antibodies. c The results of the clonogenic assay. Representative images of the cells treated the indicated conditions (left panel). The number of colonies was measured (right panel). d The sphere forming assay was performed ( > 110 sphere colonies per data point). Scale bar = 100 μm. Representative images of sphere forming assay. e PP2A-Aα/β proteins were immunoprecipitated using an anti-PP2A-Aα/β antibody and analyzed for PP2A activity. The data represent typical results and are presented as the mean ± standard deviation of three independent experiments; ** P

    Article Snippet: Briefly, CD44+ /CD24− subpopulations from MCF7 cells were isolated by their surface markers using a FACS Aria II (BD Biosciences, San Diego, CA).

    Techniques: Transfection, Irradiation, FACS, Clonogenic Assay, Immunoprecipitation, Activity Assay, Standard Deviation

    MASTL depletion does not affect the viability of normal cells. HUVEC, HDFn, MCF7, and MCF10A cells were transfected with either control siRNA or MASTL.5 siRNA for 48 h. a , f The cells were analyzed by immunoblotting with the indicated antibodies. b , c , g Cell viability was determined by WST-8 assay. d , e , h , i The cell cycle was analyzed by FACS. The data represent typical results and are presented as the mean ± standard deviation of three independent experiments; ** P

    Journal: BMC Cancer

    Article Title: MASTL inhibition promotes mitotic catastrophe through PP2A activation to inhibit cancer growth and radioresistance in breast cancer cells

    doi: 10.1186/s12885-018-4600-6

    Figure Lengend Snippet: MASTL depletion does not affect the viability of normal cells. HUVEC, HDFn, MCF7, and MCF10A cells were transfected with either control siRNA or MASTL.5 siRNA for 48 h. a , f The cells were analyzed by immunoblotting with the indicated antibodies. b , c , g Cell viability was determined by WST-8 assay. d , e , h , i The cell cycle was analyzed by FACS. The data represent typical results and are presented as the mean ± standard deviation of three independent experiments; ** P

    Article Snippet: Briefly, CD44+ /CD24− subpopulations from MCF7 cells were isolated by their surface markers using a FACS Aria II (BD Biosciences, San Diego, CA).

    Techniques: Transfection, FACS, Standard Deviation

    FGF1 is necessary for overnutrition-induced β-cell differentiation. A : RT-PCR analysis of fgf1 , insa , gcga , and amy2a expression in FACS-sorted β-cells. nc, no template control. B : Overnutrition-induced mild hyperglycemia in fgf1 mu1/mu1 fish. Wild-type and fgf1 mu1/mu1 larvae were cultured for 8 h in nutrient-free or 5% egg yolk solution at 6 dpf, and their total free glucose levels were determined immediately after. n = 10. Representative confocal projections of β-cells of 6-dpf ( C ) and 4-week-old ( E ) Tg(−1.2ins:H2B-mCherry) or fgf1 mu1/mu1 ;Tg(−1.2ins:H2B-mCherry) larvae cultured for 8 h in nutrient-free medium or 5% egg yolk. Scale bar indicates 10 µm in C and 20 μm in E . Quantification of β-cell number from 6-dpf larvae ( D ) or 4-week-old fish ( F ) suggested a loss of overnutrition-induced β-cell differentiation in fgf1 -deficient fish. n = 7–24 in D and n = 10 in F . * P

    Journal: Diabetes

    Article Title: FGF1 Mediates Overnutrition-Induced Compensatory β-Cell Differentiation

    doi: 10.2337/db15-0085

    Figure Lengend Snippet: FGF1 is necessary for overnutrition-induced β-cell differentiation. A : RT-PCR analysis of fgf1 , insa , gcga , and amy2a expression in FACS-sorted β-cells. nc, no template control. B : Overnutrition-induced mild hyperglycemia in fgf1 mu1/mu1 fish. Wild-type and fgf1 mu1/mu1 larvae were cultured for 8 h in nutrient-free or 5% egg yolk solution at 6 dpf, and their total free glucose levels were determined immediately after. n = 10. Representative confocal projections of β-cells of 6-dpf ( C ) and 4-week-old ( E ) Tg(−1.2ins:H2B-mCherry) or fgf1 mu1/mu1 ;Tg(−1.2ins:H2B-mCherry) larvae cultured for 8 h in nutrient-free medium or 5% egg yolk. Scale bar indicates 10 µm in C and 20 μm in E . Quantification of β-cell number from 6-dpf larvae ( D ) or 4-week-old fish ( F ) suggested a loss of overnutrition-induced β-cell differentiation in fgf1 -deficient fish. n = 7–24 in D and n = 10 in F . * P

    Article Snippet: The mCherry-positive β-cells were collected using the BD FACS Aria II in the Vanderbilt University Medical Center (VUMC) Flow Cytometry Core.

    Techniques: Cell Differentiation, Reverse Transcription Polymerase Chain Reaction, Expressing, FACS, Fluorescence In Situ Hybridization, Cell Culture

    Coral cell differentiation patterns. P. damicornis cell suspension was incubated with different fluorescent markers and analyzed on FACS Accuri C6. All of the samples were analyzed on two dimensional plots of the green channel- 488 nm (533/30BP) and on the far-red channel-640 nm (675/25BP) to differentiate the Symbiodinium positive cells. Gated in black are the differentiated populations with their cell percentage indicated. The analysis was done after differentiation of cells and debris on FSC and SSC and live/dead cells using PI. a ) Background control of unstained cells for all of the panels. In the gate are the Symbiodinium cells positive at 640 nm (675/25BP) due to natural fluorescence. b ) Labeled cells with Phen Green, a color that is quenched by bi-valent metals. A small population of cells had the lowest level of metal and it differentiated from the main population. c ) Labeled cells with ALDEFLUOR that indicates high levels of ALDH activity. No clear differentiation was found, but the width of the signal in different cells is 3 fluorescent log scales which enables us to differentiate low, moderate and high labeling. d ) Cells containing ROS, labeled with CellRox. Clear negative and positive populations (positive marked) were identified. Note that all the Symbiodinium positive cells are positive to ROS. e ) Cells were incubated with glucose analog 2-NBDG. Positive labeling was found, but no clear differentiation. f ) Cells were co-incubated for 3 h with green fluorescent beads for analysis of engulfment. Cells positive to beads are marked

    Journal: BMC Cell Biology

    Article Title: Coral cell separation and isolation by fluorescence-activated cell sorting (FACS)

    doi: 10.1186/s12860-017-0146-8

    Figure Lengend Snippet: Coral cell differentiation patterns. P. damicornis cell suspension was incubated with different fluorescent markers and analyzed on FACS Accuri C6. All of the samples were analyzed on two dimensional plots of the green channel- 488 nm (533/30BP) and on the far-red channel-640 nm (675/25BP) to differentiate the Symbiodinium positive cells. Gated in black are the differentiated populations with their cell percentage indicated. The analysis was done after differentiation of cells and debris on FSC and SSC and live/dead cells using PI. a ) Background control of unstained cells for all of the panels. In the gate are the Symbiodinium cells positive at 640 nm (675/25BP) due to natural fluorescence. b ) Labeled cells with Phen Green, a color that is quenched by bi-valent metals. A small population of cells had the lowest level of metal and it differentiated from the main population. c ) Labeled cells with ALDEFLUOR that indicates high levels of ALDH activity. No clear differentiation was found, but the width of the signal in different cells is 3 fluorescent log scales which enables us to differentiate low, moderate and high labeling. d ) Cells containing ROS, labeled with CellRox. Clear negative and positive populations (positive marked) were identified. Note that all the Symbiodinium positive cells are positive to ROS. e ) Cells were incubated with glucose analog 2-NBDG. Positive labeling was found, but no clear differentiation. f ) Cells were co-incubated for 3 h with green fluorescent beads for analysis of engulfment. Cells positive to beads are marked

    Article Snippet: The FACS reading was done either on BD FACS Aria II, BD LSRFortessa, BD FACS Accuri C6 (Becton, Dickinson Biosciences) or SH800S Cell Sorter (Sony).

    Techniques: Cell Differentiation, Incubation, FACS, Fluorescence, Labeling, Activity Assay

    Examples of cell differentiation from a chemical compound screening. The P. damicornis cell suspension was incubated with the indicated chemical compounds for 30 min at 20 o C and analyzed by FACS LSRFortessa. The analysis was done after differentiation of cells and debris on FSC and SSC and live/dead cells using DAPI, and separated to Symbiodinium positive cells ( e – h ) and Symbiodinium negative cells ( a – d , as shown in Fig. 1 ). a ) Background control of unstained Symbiodinium negative cells for ( b – d) . b ) Labeled cells with CAY10455, had separation for 3 potential populations shown in gates. c ) Labeled cells with Rhodamine Phenylglyoxal had separation of 4 potential populations. d ) Labeled cells with Acridine Orange had separation of 3 populations shown in red. e ) Background control of unstained Symbiodinium positive cells for ( f) . f ) Labeled cells with Dihydroethidium had separation of 3 populations positive to Symbiodinium . g ) Background control of unstained Symbiodinium positive cells for ( h ). h ) Labeled cells with Acridine Orange had separation of 3 populations positive to Symbiodinium

    Journal: BMC Cell Biology

    Article Title: Coral cell separation and isolation by fluorescence-activated cell sorting (FACS)

    doi: 10.1186/s12860-017-0146-8

    Figure Lengend Snippet: Examples of cell differentiation from a chemical compound screening. The P. damicornis cell suspension was incubated with the indicated chemical compounds for 30 min at 20 o C and analyzed by FACS LSRFortessa. The analysis was done after differentiation of cells and debris on FSC and SSC and live/dead cells using DAPI, and separated to Symbiodinium positive cells ( e – h ) and Symbiodinium negative cells ( a – d , as shown in Fig. 1 ). a ) Background control of unstained Symbiodinium negative cells for ( b – d) . b ) Labeled cells with CAY10455, had separation for 3 potential populations shown in gates. c ) Labeled cells with Rhodamine Phenylglyoxal had separation of 4 potential populations. d ) Labeled cells with Acridine Orange had separation of 3 populations shown in red. e ) Background control of unstained Symbiodinium positive cells for ( f) . f ) Labeled cells with Dihydroethidium had separation of 3 populations positive to Symbiodinium . g ) Background control of unstained Symbiodinium positive cells for ( h ). h ) Labeled cells with Acridine Orange had separation of 3 populations positive to Symbiodinium

    Article Snippet: The FACS reading was done either on BD FACS Aria II, BD LSRFortessa, BD FACS Accuri C6 (Becton, Dickinson Biosciences) or SH800S Cell Sorter (Sony).

    Techniques: Cell Differentiation, Incubation, FACS, Labeling

    Fluorescence analysis of 2-NBDG uptake by flow cytometry. FACS analysis of 2-NBDG uptake in differentiated L6 cells by plotting cell count against FITC revealed that 8%, 8.1% and 30% of cells uptake 2-NBDG in control, TBHP and Rosiglitazone treated cells respectively whereas 30.6%, 33.1%, 28%, 32% of cells uptake 2-NBDG, pretreated with two different concentrations (10 and 100 μM) of Naringin along with/without TBHP respectively. Each value represents mean ± SD (standard deviation) from triplicate measurements (n = 3) of three different experiments. Significance test between various groups was determined by using one way ANOVA followed by Duncan’s multiple range test. * P≤0.05 versus control.

    Journal: PLoS ONE

    Article Title: Preconditioning L6 Muscle Cells with Naringin Ameliorates Oxidative Stress and Increases Glucose Uptake

    doi: 10.1371/journal.pone.0132429

    Figure Lengend Snippet: Fluorescence analysis of 2-NBDG uptake by flow cytometry. FACS analysis of 2-NBDG uptake in differentiated L6 cells by plotting cell count against FITC revealed that 8%, 8.1% and 30% of cells uptake 2-NBDG in control, TBHP and Rosiglitazone treated cells respectively whereas 30.6%, 33.1%, 28%, 32% of cells uptake 2-NBDG, pretreated with two different concentrations (10 and 100 μM) of Naringin along with/without TBHP respectively. Each value represents mean ± SD (standard deviation) from triplicate measurements (n = 3) of three different experiments. Significance test between various groups was determined by using one way ANOVA followed by Duncan’s multiple range test. * P≤0.05 versus control.

    Article Snippet: Samples were analyzed using BD FACS Aria II (BD Biosciences) at FITC range (excitation 490 nm, emission 525 nm band pass filter).

    Techniques: Fluorescence, Flow Cytometry, Cytometry, FACS, Cell Counting, Standard Deviation