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cytoflex s flow cytometry  (Danaher Inc)


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    Danaher Inc cytoflex s flow cytometry
    (A) Summary of gene deletion vs. gene overexpression approaches. (B) Rationale for the present study. (C) Experimental outline of the present study. (D) HEK293 and Jurkat cells were inoculated with different volumes of Ebola or rabies pseudovirus encoding a cell-surface marker (mCD19t; a truncated mutant of mouse CD19), followed by flow <t>cytometry</t> to determine the percentage of infected cells. This revealed that Jurkat cells are largely refractory to Ebola or rabies pseudovirus entry, relative to HEK293 cells. (E) Construction of a clonal Jurkat cell line, known as “Jurkat C6”, stably expressing a degron-tagged CRISPRa construct ( left ). Jurkat C6 cells were transduced with sgRNA targeting the endogenous human CD19 gene, in the presence or absence of TMP (1 μM) for 3 days, followed by flow cytometry to detect hCD19 expression ( right ). (F) At each of the indicated points of the genome-wide CRISPRa screen, the cell population was challenged with Ebola or rabies pseudovirus, and cell infectivity was evaluated by flow cytometry. Upon successive rounds of the screen, the cell population became progressively more susceptible to infection to either Ebola or rabies pseudovirus, respectively. (G) sgRNA distribution upon successive rounds of the genome-wide CRISPRa screen for Ebola pseudovirus entry. (H) sgRNA distribution upon successive rounds of the genome-wide CRISPRa screen for rabies pseudovirus entry.
    Cytoflex S Flow Cytometry, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 97/100, based on 3258 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cytoflex+s+flow+cytometry/CytoFLEX+S+Flow+Cytometer/bio_rxiv__64898__2026__03__06__710083-191-21-25
    Average 97 stars, based on 3258 article reviews
    cytoflex s flow cytometry - by Bioz Stars, 2026-09
    97/100 stars

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    1) Product Images from "Elucidating genes sufficient for viral entry into cells through sequential genome-wide CRISPR activation screens"

    Article Title: Elucidating genes sufficient for viral entry into cells through sequential genome-wide CRISPR activation screens

    Journal: bioRxiv

    doi: 10.64898/2026.03.06.710083

    (A) Summary of gene deletion vs. gene overexpression approaches. (B) Rationale for the present study. (C) Experimental outline of the present study. (D) HEK293 and Jurkat cells were inoculated with different volumes of Ebola or rabies pseudovirus encoding a cell-surface marker (mCD19t; a truncated mutant of mouse CD19), followed by flow cytometry to determine the percentage of infected cells. This revealed that Jurkat cells are largely refractory to Ebola or rabies pseudovirus entry, relative to HEK293 cells. (E) Construction of a clonal Jurkat cell line, known as “Jurkat C6”, stably expressing a degron-tagged CRISPRa construct ( left ). Jurkat C6 cells were transduced with sgRNA targeting the endogenous human CD19 gene, in the presence or absence of TMP (1 μM) for 3 days, followed by flow cytometry to detect hCD19 expression ( right ). (F) At each of the indicated points of the genome-wide CRISPRa screen, the cell population was challenged with Ebola or rabies pseudovirus, and cell infectivity was evaluated by flow cytometry. Upon successive rounds of the screen, the cell population became progressively more susceptible to infection to either Ebola or rabies pseudovirus, respectively. (G) sgRNA distribution upon successive rounds of the genome-wide CRISPRa screen for Ebola pseudovirus entry. (H) sgRNA distribution upon successive rounds of the genome-wide CRISPRa screen for rabies pseudovirus entry.
    Figure Legend Snippet: (A) Summary of gene deletion vs. gene overexpression approaches. (B) Rationale for the present study. (C) Experimental outline of the present study. (D) HEK293 and Jurkat cells were inoculated with different volumes of Ebola or rabies pseudovirus encoding a cell-surface marker (mCD19t; a truncated mutant of mouse CD19), followed by flow cytometry to determine the percentage of infected cells. This revealed that Jurkat cells are largely refractory to Ebola or rabies pseudovirus entry, relative to HEK293 cells. (E) Construction of a clonal Jurkat cell line, known as “Jurkat C6”, stably expressing a degron-tagged CRISPRa construct ( left ). Jurkat C6 cells were transduced with sgRNA targeting the endogenous human CD19 gene, in the presence or absence of TMP (1 μM) for 3 days, followed by flow cytometry to detect hCD19 expression ( right ). (F) At each of the indicated points of the genome-wide CRISPRa screen, the cell population was challenged with Ebola or rabies pseudovirus, and cell infectivity was evaluated by flow cytometry. Upon successive rounds of the screen, the cell population became progressively more susceptible to infection to either Ebola or rabies pseudovirus, respectively. (G) sgRNA distribution upon successive rounds of the genome-wide CRISPRa screen for Ebola pseudovirus entry. (H) sgRNA distribution upon successive rounds of the genome-wide CRISPRa screen for rabies pseudovirus entry.

    Techniques Used: Over Expression, Marker, Mutagenesis, Flow Cytometry, Infection, Stable Transfection, Expressing, Construct, Transduction, Genome Wide

    A) Plasmids used to pseudotype non-replicating lentiviruses with either Ebola or rabies envelope proteins. EBOV-GP: glycoprotein of Ebola virus, Makona variant. RABV-GP N2C: glycoprotein of rabies virus, N2C variant. B) HEK293 and Jurkat cells were inoculated with different volumes of VSV envelope protein-pseudotyped lentivirus encoding a cell-surface marker (mCD19t; a truncated mutant of mouse CD19), followed by flow cytometry to determine the percentage of infected cells. This served as a positive control to confirm that Jurkat cells and HEK293 cells are both susceptible to VSV pseudovirus entry. C) Jurkat C6 cells were transduced with sgRNA targeting the endogenous human CD19 gene, in the presence of different TMP concentrations (0-4 μM) for 2-3 days, followed by flow cytometry to detect human CD19 expression.
    Figure Legend Snippet: A) Plasmids used to pseudotype non-replicating lentiviruses with either Ebola or rabies envelope proteins. EBOV-GP: glycoprotein of Ebola virus, Makona variant. RABV-GP N2C: glycoprotein of rabies virus, N2C variant. B) HEK293 and Jurkat cells were inoculated with different volumes of VSV envelope protein-pseudotyped lentivirus encoding a cell-surface marker (mCD19t; a truncated mutant of mouse CD19), followed by flow cytometry to determine the percentage of infected cells. This served as a positive control to confirm that Jurkat cells and HEK293 cells are both susceptible to VSV pseudovirus entry. C) Jurkat C6 cells were transduced with sgRNA targeting the endogenous human CD19 gene, in the presence of different TMP concentrations (0-4 μM) for 2-3 days, followed by flow cytometry to detect human CD19 expression.

    Techniques Used: Virus, Variant Assay, Marker, Mutagenesis, Flow Cytometry, Infection, Positive Control, Transduction, Expressing

    A) NGFR was expressed in Jurkat C6 cells using CRISPRa, or alternatively, Jurkat cells using cDNA expression. NGFR -expressing or control cells were then inoculated with rabies pseudovirus encoding mCD19t. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that NGFR expression significantly increased the susceptibility of Jurkat cells to rabies pseudovirus infection. B) L-SIGN or DC-SIGN were expressed in Jurkat or primary human CD4 + T cells using cDNA expression. L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with rabies pseudovirus encoding hEGFRt (a truncated mutant of human EGFR). Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of Jurkat cells and primary T cells to Ebola pseudovirus infection. C) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression, and then L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with GFP -expressing Ebola virus or zsGreen -expressing Sudan virus under BSL4 containment. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of primary T cells to authentic Ebola and Sudan virus infection.
    Figure Legend Snippet: A) NGFR was expressed in Jurkat C6 cells using CRISPRa, or alternatively, Jurkat cells using cDNA expression. NGFR -expressing or control cells were then inoculated with rabies pseudovirus encoding mCD19t. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that NGFR expression significantly increased the susceptibility of Jurkat cells to rabies pseudovirus infection. B) L-SIGN or DC-SIGN were expressed in Jurkat or primary human CD4 + T cells using cDNA expression. L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with rabies pseudovirus encoding hEGFRt (a truncated mutant of human EGFR). Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of Jurkat cells and primary T cells to Ebola pseudovirus infection. C) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression, and then L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with GFP -expressing Ebola virus or zsGreen -expressing Sudan virus under BSL4 containment. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of primary T cells to authentic Ebola and Sudan virus infection.

    Techniques Used: Expressing, Control, Flow Cytometry, Infection, Mutagenesis, Virus

    A) NGFR was expressed in Jurkat C6 cells using CRISPRa, or alternatively, Jurkat cells using cDNA expression. NGFR -expressing or control cells were then inoculated with rabies pseudovirus encoding mCD19t. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that NGFR expression significantly increased the susceptibility of Jurkat cells to rabies pseudovirus infection. As positive controls, flow cytometry was used to confirm successful delivery of the sgRNA construct as part of the CRISPRa workflow (as denoted by BFP expression) and that NGFR was expressed (upon cDNA expression). B) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression. L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with rabies pseudovirus encoding hEGFRt (a truncated mutant of human EGFR). Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of Jurkat cells and primary T cells to Ebola pseudovirus infection. Cells expressing the highest levels of L-SIGN and DC-SIGN were preferentially infected by Ebola pseudovirus. C) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression, and then L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with GFP -expressing Ebola virus or zsGreen -expressing Sudan virus under BSL4 containment. On days 0, 1, and 2 post-infection, flow cytometry was performed to determine the percentage of infected cells and qPCR was performed on cell culture supernatants to quantify viral genome replication. This revealed that L-SIGN or DC-SIGN expression enabled authentic Ebola and Sudan virus entry into primary human T cells, but viral genome replication was impaired, perhaps reflective of cell-intrinsic restriction factors.
    Figure Legend Snippet: A) NGFR was expressed in Jurkat C6 cells using CRISPRa, or alternatively, Jurkat cells using cDNA expression. NGFR -expressing or control cells were then inoculated with rabies pseudovirus encoding mCD19t. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that NGFR expression significantly increased the susceptibility of Jurkat cells to rabies pseudovirus infection. As positive controls, flow cytometry was used to confirm successful delivery of the sgRNA construct as part of the CRISPRa workflow (as denoted by BFP expression) and that NGFR was expressed (upon cDNA expression). B) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression. L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with rabies pseudovirus encoding hEGFRt (a truncated mutant of human EGFR). Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of Jurkat cells and primary T cells to Ebola pseudovirus infection. Cells expressing the highest levels of L-SIGN and DC-SIGN were preferentially infected by Ebola pseudovirus. C) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression, and then L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with GFP -expressing Ebola virus or zsGreen -expressing Sudan virus under BSL4 containment. On days 0, 1, and 2 post-infection, flow cytometry was performed to determine the percentage of infected cells and qPCR was performed on cell culture supernatants to quantify viral genome replication. This revealed that L-SIGN or DC-SIGN expression enabled authentic Ebola and Sudan virus entry into primary human T cells, but viral genome replication was impaired, perhaps reflective of cell-intrinsic restriction factors.

    Techniques Used: Expressing, Control, Flow Cytometry, Infection, Construct, Mutagenesis, Virus, Cell Culture

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    Fluorescence:

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    Identification of a fluorescent probe for spatial tracing. (A) Staining patterns using different fluorescent probes. H1 hPSCs were cultured in E8 medium on the Matrigel-coated surface for 3 days before they were stained and observed under a fluorescence microscope. (B) The stained cells were analyzed by flow <t>cytometry.</t> (C) Staining patterns during individualization. After hPSC colonies were stained using JC-1 dye, they were dissociated using TrypLE and observed under a microscope before cell collection. (D) JC-1 stain after cell sorting. After hPSC colonies were stained with JC-1, cells were individualized with TryPLE, sorted by flow cytometry, and the two populations seeded back onto the Matrigel-coated surface. Cell morphology and fluorescent signals were observed 6 hours after plating.
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    ( A ) Flow <t>cytometry</t> staining of HepG 2 cells with apoptosis and necrosis indicators Apopxin Red and Nuclear Green DSC1, respectively, following 6 hr incubation with 150 µg/mL copper chalcogenide NCs (cation concentration). ( B ). Summary and statistical analysis of the necrosis assay (n = 3). * p < 0.05, **** p < 0.0001.
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    (A) Summary of gene deletion vs. gene overexpression approaches. (B) Rationale for the present study. (C) Experimental outline of the present study. (D) HEK293 and Jurkat cells were inoculated with different volumes of Ebola or rabies pseudovirus encoding a cell-surface marker (mCD19t; a truncated mutant of mouse CD19), followed by flow cytometry to determine the percentage of infected cells. This revealed that Jurkat cells are largely refractory to Ebola or rabies pseudovirus entry, relative to HEK293 cells. (E) Construction of a clonal Jurkat cell line, known as “Jurkat C6”, stably expressing a degron-tagged CRISPRa construct ( left ). Jurkat C6 cells were transduced with sgRNA targeting the endogenous human CD19 gene, in the presence or absence of TMP (1 μM) for 3 days, followed by flow cytometry to detect hCD19 expression ( right ). (F) At each of the indicated points of the genome-wide CRISPRa screen, the cell population was challenged with Ebola or rabies pseudovirus, and cell infectivity was evaluated by flow cytometry. Upon successive rounds of the screen, the cell population became progressively more susceptible to infection to either Ebola or rabies pseudovirus, respectively. (G) sgRNA distribution upon successive rounds of the genome-wide CRISPRa screen for Ebola pseudovirus entry. (H) sgRNA distribution upon successive rounds of the genome-wide CRISPRa screen for rabies pseudovirus entry.

    Journal: bioRxiv

    Article Title: Elucidating genes sufficient for viral entry into cells through sequential genome-wide CRISPR activation screens

    doi: 10.64898/2026.03.06.710083

    Figure Lengend Snippet: (A) Summary of gene deletion vs. gene overexpression approaches. (B) Rationale for the present study. (C) Experimental outline of the present study. (D) HEK293 and Jurkat cells were inoculated with different volumes of Ebola or rabies pseudovirus encoding a cell-surface marker (mCD19t; a truncated mutant of mouse CD19), followed by flow cytometry to determine the percentage of infected cells. This revealed that Jurkat cells are largely refractory to Ebola or rabies pseudovirus entry, relative to HEK293 cells. (E) Construction of a clonal Jurkat cell line, known as “Jurkat C6”, stably expressing a degron-tagged CRISPRa construct ( left ). Jurkat C6 cells were transduced with sgRNA targeting the endogenous human CD19 gene, in the presence or absence of TMP (1 μM) for 3 days, followed by flow cytometry to detect hCD19 expression ( right ). (F) At each of the indicated points of the genome-wide CRISPRa screen, the cell population was challenged with Ebola or rabies pseudovirus, and cell infectivity was evaluated by flow cytometry. Upon successive rounds of the screen, the cell population became progressively more susceptible to infection to either Ebola or rabies pseudovirus, respectively. (G) sgRNA distribution upon successive rounds of the genome-wide CRISPRa screen for Ebola pseudovirus entry. (H) sgRNA distribution upon successive rounds of the genome-wide CRISPRa screen for rabies pseudovirus entry.

    Article Snippet: Afterwards, cells were centrifuged at 1000g for 10 minutes, resuspended in FACS buffer, and flow cytometric measurements were performed on a CytoFLEX S flow cytometry (Beckman Coulter).

    Techniques: Over Expression, Marker, Mutagenesis, Flow Cytometry, Infection, Stable Transfection, Expressing, Construct, Transduction, Genome Wide

    A) Plasmids used to pseudotype non-replicating lentiviruses with either Ebola or rabies envelope proteins. EBOV-GP: glycoprotein of Ebola virus, Makona variant. RABV-GP N2C: glycoprotein of rabies virus, N2C variant. B) HEK293 and Jurkat cells were inoculated with different volumes of VSV envelope protein-pseudotyped lentivirus encoding a cell-surface marker (mCD19t; a truncated mutant of mouse CD19), followed by flow cytometry to determine the percentage of infected cells. This served as a positive control to confirm that Jurkat cells and HEK293 cells are both susceptible to VSV pseudovirus entry. C) Jurkat C6 cells were transduced with sgRNA targeting the endogenous human CD19 gene, in the presence of different TMP concentrations (0-4 μM) for 2-3 days, followed by flow cytometry to detect human CD19 expression.

    Journal: bioRxiv

    Article Title: Elucidating genes sufficient for viral entry into cells through sequential genome-wide CRISPR activation screens

    doi: 10.64898/2026.03.06.710083

    Figure Lengend Snippet: A) Plasmids used to pseudotype non-replicating lentiviruses with either Ebola or rabies envelope proteins. EBOV-GP: glycoprotein of Ebola virus, Makona variant. RABV-GP N2C: glycoprotein of rabies virus, N2C variant. B) HEK293 and Jurkat cells were inoculated with different volumes of VSV envelope protein-pseudotyped lentivirus encoding a cell-surface marker (mCD19t; a truncated mutant of mouse CD19), followed by flow cytometry to determine the percentage of infected cells. This served as a positive control to confirm that Jurkat cells and HEK293 cells are both susceptible to VSV pseudovirus entry. C) Jurkat C6 cells were transduced with sgRNA targeting the endogenous human CD19 gene, in the presence of different TMP concentrations (0-4 μM) for 2-3 days, followed by flow cytometry to detect human CD19 expression.

    Article Snippet: Afterwards, cells were centrifuged at 1000g for 10 minutes, resuspended in FACS buffer, and flow cytometric measurements were performed on a CytoFLEX S flow cytometry (Beckman Coulter).

    Techniques: Virus, Variant Assay, Marker, Mutagenesis, Flow Cytometry, Infection, Positive Control, Transduction, Expressing

    A) NGFR was expressed in Jurkat C6 cells using CRISPRa, or alternatively, Jurkat cells using cDNA expression. NGFR -expressing or control cells were then inoculated with rabies pseudovirus encoding mCD19t. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that NGFR expression significantly increased the susceptibility of Jurkat cells to rabies pseudovirus infection. B) L-SIGN or DC-SIGN were expressed in Jurkat or primary human CD4 + T cells using cDNA expression. L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with rabies pseudovirus encoding hEGFRt (a truncated mutant of human EGFR). Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of Jurkat cells and primary T cells to Ebola pseudovirus infection. C) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression, and then L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with GFP -expressing Ebola virus or zsGreen -expressing Sudan virus under BSL4 containment. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of primary T cells to authentic Ebola and Sudan virus infection.

    Journal: bioRxiv

    Article Title: Elucidating genes sufficient for viral entry into cells through sequential genome-wide CRISPR activation screens

    doi: 10.64898/2026.03.06.710083

    Figure Lengend Snippet: A) NGFR was expressed in Jurkat C6 cells using CRISPRa, or alternatively, Jurkat cells using cDNA expression. NGFR -expressing or control cells were then inoculated with rabies pseudovirus encoding mCD19t. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that NGFR expression significantly increased the susceptibility of Jurkat cells to rabies pseudovirus infection. B) L-SIGN or DC-SIGN were expressed in Jurkat or primary human CD4 + T cells using cDNA expression. L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with rabies pseudovirus encoding hEGFRt (a truncated mutant of human EGFR). Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of Jurkat cells and primary T cells to Ebola pseudovirus infection. C) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression, and then L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with GFP -expressing Ebola virus or zsGreen -expressing Sudan virus under BSL4 containment. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of primary T cells to authentic Ebola and Sudan virus infection.

    Article Snippet: Afterwards, cells were centrifuged at 1000g for 10 minutes, resuspended in FACS buffer, and flow cytometric measurements were performed on a CytoFLEX S flow cytometry (Beckman Coulter).

    Techniques: Expressing, Control, Flow Cytometry, Infection, Mutagenesis, Virus

    A) NGFR was expressed in Jurkat C6 cells using CRISPRa, or alternatively, Jurkat cells using cDNA expression. NGFR -expressing or control cells were then inoculated with rabies pseudovirus encoding mCD19t. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that NGFR expression significantly increased the susceptibility of Jurkat cells to rabies pseudovirus infection. As positive controls, flow cytometry was used to confirm successful delivery of the sgRNA construct as part of the CRISPRa workflow (as denoted by BFP expression) and that NGFR was expressed (upon cDNA expression). B) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression. L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with rabies pseudovirus encoding hEGFRt (a truncated mutant of human EGFR). Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of Jurkat cells and primary T cells to Ebola pseudovirus infection. Cells expressing the highest levels of L-SIGN and DC-SIGN were preferentially infected by Ebola pseudovirus. C) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression, and then L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with GFP -expressing Ebola virus or zsGreen -expressing Sudan virus under BSL4 containment. On days 0, 1, and 2 post-infection, flow cytometry was performed to determine the percentage of infected cells and qPCR was performed on cell culture supernatants to quantify viral genome replication. This revealed that L-SIGN or DC-SIGN expression enabled authentic Ebola and Sudan virus entry into primary human T cells, but viral genome replication was impaired, perhaps reflective of cell-intrinsic restriction factors.

    Journal: bioRxiv

    Article Title: Elucidating genes sufficient for viral entry into cells through sequential genome-wide CRISPR activation screens

    doi: 10.64898/2026.03.06.710083

    Figure Lengend Snippet: A) NGFR was expressed in Jurkat C6 cells using CRISPRa, or alternatively, Jurkat cells using cDNA expression. NGFR -expressing or control cells were then inoculated with rabies pseudovirus encoding mCD19t. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that NGFR expression significantly increased the susceptibility of Jurkat cells to rabies pseudovirus infection. As positive controls, flow cytometry was used to confirm successful delivery of the sgRNA construct as part of the CRISPRa workflow (as denoted by BFP expression) and that NGFR was expressed (upon cDNA expression). B) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression. L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with rabies pseudovirus encoding hEGFRt (a truncated mutant of human EGFR). Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of Jurkat cells and primary T cells to Ebola pseudovirus infection. Cells expressing the highest levels of L-SIGN and DC-SIGN were preferentially infected by Ebola pseudovirus. C) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression, and then L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with GFP -expressing Ebola virus or zsGreen -expressing Sudan virus under BSL4 containment. On days 0, 1, and 2 post-infection, flow cytometry was performed to determine the percentage of infected cells and qPCR was performed on cell culture supernatants to quantify viral genome replication. This revealed that L-SIGN or DC-SIGN expression enabled authentic Ebola and Sudan virus entry into primary human T cells, but viral genome replication was impaired, perhaps reflective of cell-intrinsic restriction factors.

    Article Snippet: Afterwards, cells were centrifuged at 1000g for 10 minutes, resuspended in FACS buffer, and flow cytometric measurements were performed on a CytoFLEX S flow cytometry (Beckman Coulter).

    Techniques: Expressing, Control, Flow Cytometry, Infection, Construct, Mutagenesis, Virus, Cell Culture

    Identification of a fluorescent probe for spatial tracing. (A) Staining patterns using different fluorescent probes. H1 hPSCs were cultured in E8 medium on the Matrigel-coated surface for 3 days before they were stained and observed under a fluorescence microscope. (B) The stained cells were analyzed by flow cytometry. (C) Staining patterns during individualization. After hPSC colonies were stained using JC-1 dye, they were dissociated using TrypLE and observed under a microscope before cell collection. (D) JC-1 stain after cell sorting. After hPSC colonies were stained with JC-1, cells were individualized with TryPLE, sorted by flow cytometry, and the two populations seeded back onto the Matrigel-coated surface. Cell morphology and fluorescent signals were observed 6 hours after plating.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Metabolic patterns predispose human pluripotent stem cells to spatial organization of cell fate

    doi: 10.3389/fcell.2025.1696372

    Figure Lengend Snippet: Identification of a fluorescent probe for spatial tracing. (A) Staining patterns using different fluorescent probes. H1 hPSCs were cultured in E8 medium on the Matrigel-coated surface for 3 days before they were stained and observed under a fluorescence microscope. (B) The stained cells were analyzed by flow cytometry. (C) Staining patterns during individualization. After hPSC colonies were stained using JC-1 dye, they were dissociated using TrypLE and observed under a microscope before cell collection. (D) JC-1 stain after cell sorting. After hPSC colonies were stained with JC-1, cells were individualized with TryPLE, sorted by flow cytometry, and the two populations seeded back onto the Matrigel-coated surface. Cell morphology and fluorescent signals were observed 6 hours after plating.

    Article Snippet: Stained cells were observed in fresh culture medium under an EVOS FL Auto fluorescence microscope before cells were individualized using TrypLE Select for flow cytometry analysis (Beckman Coulter CytoFLEX S flow cytometer) or cell sorting (BD FACSAriaTM III cell sorter).

    Techniques: Staining, Cell Culture, Fluorescence, Microscopy, Flow Cytometry, FACS

    Spatial patterns of hPSCs on the Matrigel-coated surface during maintenance and differentiation. (A–C) Time course of JC-1 pattern changes during maintenance. hPSCs were passaged onto the Matrigel-coated surface using DPBS-EDTA, and they were stained with JC-1 daily before analysis by microscopy (A) and flow cytometry (B) . Flow cytometry data are presented as mean ± SD of three biological replicates (C) . ImageJ was used to measure the width of the JC-1-positive ring in hPSC colonies. n = 20 data points on the image. (D) Comparison of nuclear staining (Hoechst) and JC-1 staining in hPSC colonies to show the width of the ring. (E) JC-1-based cell tracing during maintenance. hPSC colonies were stained with JC-1 briefly and were cultured and observed in E8 medium. (F) JC-1-based cell tracing during BMP4-induced differentiation. hPSC colonies were stained with JC-1 briefly and were cultured and observed in E8 medium with BMP4 (20 ng/mL).

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Metabolic patterns predispose human pluripotent stem cells to spatial organization of cell fate

    doi: 10.3389/fcell.2025.1696372

    Figure Lengend Snippet: Spatial patterns of hPSCs on the Matrigel-coated surface during maintenance and differentiation. (A–C) Time course of JC-1 pattern changes during maintenance. hPSCs were passaged onto the Matrigel-coated surface using DPBS-EDTA, and they were stained with JC-1 daily before analysis by microscopy (A) and flow cytometry (B) . Flow cytometry data are presented as mean ± SD of three biological replicates (C) . ImageJ was used to measure the width of the JC-1-positive ring in hPSC colonies. n = 20 data points on the image. (D) Comparison of nuclear staining (Hoechst) and JC-1 staining in hPSC colonies to show the width of the ring. (E) JC-1-based cell tracing during maintenance. hPSC colonies were stained with JC-1 briefly and were cultured and observed in E8 medium. (F) JC-1-based cell tracing during BMP4-induced differentiation. hPSC colonies were stained with JC-1 briefly and were cultured and observed in E8 medium with BMP4 (20 ng/mL).

    Article Snippet: Stained cells were observed in fresh culture medium under an EVOS FL Auto fluorescence microscope before cells were individualized using TrypLE Select for flow cytometry analysis (Beckman Coulter CytoFLEX S flow cytometer) or cell sorting (BD FACSAriaTM III cell sorter).

    Techniques: Staining, Microscopy, Flow Cytometry, Comparison, Cell Culture

    Cell adhesion-promoting factors affect metabolic and cell fate patterns. (A,B) Effect of the coating surface on the JC-1 pattern in hPSC colonies. hPSCs were plated onto a Matrigel- or vitronectin- or E-cadherin-coated surface and cultured for 2 days, before they were stained with JC-1 and analyzed using fluorescence microscopy (A) and flow cytometry (B,C) . H1 hPSC colonies were cultured on the surface containing both Matrigel and E-cadherin for 2 days and stained with JC-1. (D,E) Cell adhesion-promoting factors affected mesoderm differentiation induced by BMP4. H1 hPSCs were cultured for 2 days on surfaces coated with either Matrigel or E-cadherin, prior to a 2-day induction with BMP4. Differentiation toward mesoderm was assessed by immunostaining for the marker TBXT (D) . Mesoderm markers TBXT (T) and MIXL1 were analyzed using RT-qPCR (E) (n = 4, *P < 0.05).

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Metabolic patterns predispose human pluripotent stem cells to spatial organization of cell fate

    doi: 10.3389/fcell.2025.1696372

    Figure Lengend Snippet: Cell adhesion-promoting factors affect metabolic and cell fate patterns. (A,B) Effect of the coating surface on the JC-1 pattern in hPSC colonies. hPSCs were plated onto a Matrigel- or vitronectin- or E-cadherin-coated surface and cultured for 2 days, before they were stained with JC-1 and analyzed using fluorescence microscopy (A) and flow cytometry (B,C) . H1 hPSC colonies were cultured on the surface containing both Matrigel and E-cadherin for 2 days and stained with JC-1. (D,E) Cell adhesion-promoting factors affected mesoderm differentiation induced by BMP4. H1 hPSCs were cultured for 2 days on surfaces coated with either Matrigel or E-cadherin, prior to a 2-day induction with BMP4. Differentiation toward mesoderm was assessed by immunostaining for the marker TBXT (D) . Mesoderm markers TBXT (T) and MIXL1 were analyzed using RT-qPCR (E) (n = 4, *P < 0.05).

    Article Snippet: Stained cells were observed in fresh culture medium under an EVOS FL Auto fluorescence microscope before cells were individualized using TrypLE Select for flow cytometry analysis (Beckman Coulter CytoFLEX S flow cytometer) or cell sorting (BD FACSAriaTM III cell sorter).

    Techniques: Cell Culture, Staining, Fluorescence, Microscopy, Flow Cytometry, Immunostaining, Marker, Quantitative RT-PCR

    mTOR and ROCK modulate metabolic and cell fate pattern formation in hPSC colonies. (A,B) JC-1 patterns under mTOR and ROCK inhibition. hPSCs were passaged with 10 µM Y27632 onto Matrigel-coated surfaces in E8 medium. Cells were then treated with either 100 nM rapamycin (mTOR inhibitor) or 10 µM Y27632 (ROCK inhibitor) on day 1 and day 2. JC-1 staining and microscopy observation were performed on day 3. Cells were also stained with JC-1 and analyzed by flow cytometry daily to monitor mitochondrial membrane potential changes (B) . (C,D) TBXT immunostaining in BMP4-induced H1 hPSCs. Immunostaining of TBXT on day 4, following BMP4 induction on day 2, with or without rapamycin or Y27632 treatment. (E) RT-qPCR analysis of mesoderm induction under different treatments on day 4 of differentiation following BMP4 induction on day 2 (n = 4, *P < 0.05).

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Metabolic patterns predispose human pluripotent stem cells to spatial organization of cell fate

    doi: 10.3389/fcell.2025.1696372

    Figure Lengend Snippet: mTOR and ROCK modulate metabolic and cell fate pattern formation in hPSC colonies. (A,B) JC-1 patterns under mTOR and ROCK inhibition. hPSCs were passaged with 10 µM Y27632 onto Matrigel-coated surfaces in E8 medium. Cells were then treated with either 100 nM rapamycin (mTOR inhibitor) or 10 µM Y27632 (ROCK inhibitor) on day 1 and day 2. JC-1 staining and microscopy observation were performed on day 3. Cells were also stained with JC-1 and analyzed by flow cytometry daily to monitor mitochondrial membrane potential changes (B) . (C,D) TBXT immunostaining in BMP4-induced H1 hPSCs. Immunostaining of TBXT on day 4, following BMP4 induction on day 2, with or without rapamycin or Y27632 treatment. (E) RT-qPCR analysis of mesoderm induction under different treatments on day 4 of differentiation following BMP4 induction on day 2 (n = 4, *P < 0.05).

    Article Snippet: Stained cells were observed in fresh culture medium under an EVOS FL Auto fluorescence microscope before cells were individualized using TrypLE Select for flow cytometry analysis (Beckman Coulter CytoFLEX S flow cytometer) or cell sorting (BD FACSAriaTM III cell sorter).

    Techniques: Inhibition, Staining, Microscopy, Flow Cytometry, Membrane, Immunostaining, Quantitative RT-PCR

    ( A ) Flow cytometry staining of HepG 2 cells with apoptosis and necrosis indicators Apopxin Red and Nuclear Green DSC1, respectively, following 6 hr incubation with 150 µg/mL copper chalcogenide NCs (cation concentration). ( B ). Summary and statistical analysis of the necrosis assay (n = 3). * p < 0.05, **** p < 0.0001.

    Journal: bioRxiv

    Article Title: Elemental Composition and Degradation Rate Impact the Biocompatibility of Copper Chalcogenide Nanocrystals

    doi: 10.64898/2025.12.17.695045

    Figure Lengend Snippet: ( A ) Flow cytometry staining of HepG 2 cells with apoptosis and necrosis indicators Apopxin Red and Nuclear Green DSC1, respectively, following 6 hr incubation with 150 µg/mL copper chalcogenide NCs (cation concentration). ( B ). Summary and statistical analysis of the necrosis assay (n = 3). * p < 0.05, **** p < 0.0001.

    Article Snippet: The cells were measured using a multi-channel CytoFLEX S flow cytometry analyzer (Beckman Coulter, CA, USA).

    Techniques: Flow Cytometry, Staining, Incubation, Concentration Assay