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Aqua vital dye

Manufactured by Thermo Fisher Scientific
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Aqua Vital Dye is a fluorescent dye used for labeling and visualization of nucleic acids in aqueous solutions. The dye binds to DNA and RNA and emits fluorescence upon excitation, allowing for the detection and quantification of these biomolecules.

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Lab products found in correlation

Igd pe clone ia6 2, by BD (1 mentions) Facsaria 2, by BD (1 mentions) Cd3 pe cy5 clone hit3a, by BD (1 mentions) Cd27 pe cy7 clone o323, by Thermo Fisher Scientific (1 mentions) Facsaria 2 cell sorter, by BD (1 mentions) Cd69 fitc, by Thermo Fisher Scientific (1 mentions) Live dead fixable far red, by Thermo Fisher Scientific (1 mentions) Cd11c pe cy7 hl3, by BD (1 mentions)

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Aqua dye (13 citations)

5 protocols using aqua vital dye

1

ZIKV-Reactive B Cell Profiling

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Thawed PBMCs were stained with Aqua Vital Dye (Invitrogen), IgD-PE (clone IA6-2; BD Biosciences), CD10 PE-CF594 (clone HI10a; BD Biosciences; CD3 PE-Cy5 (clone HIT3a; BD Biosciences), CD14 BV605 (clone M5E2; BD Biolegend), CD16 BV570 (clone 3G8; Biolegend), CD27 PE-Cy7 (clone O323; Thermo Fisher Scientific), CD38 APC-AF700 (clone LS198-4-3; Beckman Coulter), CD19 APC-Cy7 (clone SJ25C1; BD Biosciences) and 1 x 106 PFU of freshly thawed UV-inactivated ZIKV labelled with AF488. Additionally, 5 μM of Chk2 kinase inhibitor II (Calbiochem/EMD Chemicals) was added to prevent cell death. Unfractionated B cells were defined as CD14−/CD16−/CD3−/CD19+. For memory B cells an additional IgD−/CD27all gate was applied. The ZIKV-reactive AF488 gate was set using a fluorescence minus one condition. Cells were sorted on a Beckton Dickinson FACS Aria II and analysis was performed in FlowJo.
Singh T., Hwang K.K., Miller A.S., Jones R.L., Lopez C.A., Dulson S.J., Giuberti C., Gladden M.A., Miller I., Webster H.S., Eudailey J.A., Luo K., Von Holle T., Edwards R.J., Valencia S., Burgomaster K.E., Zhang S., Mangold J.F., Tu J.J., Dennis M., Alam S.M., Premkumar L., Dietze R., Pierson T.C., Eong Ooi E., Lazear H.M., Kuhn R.J., Permar S.R, & Bonsignori M. (2022). A Zika virus-specific IgM elicited in pregnancy exhibits ultrapotent neutralization. Cell, 185(25), 4826-4840.e17.
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2

Characterizing Macaque B and T Cell Subsets

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Macaque PBMCs were stained by a panel of markers, including CD38, Bcl-6, CD1c, CD69, CD21, Ki67, IgG, IgM, CD3/CD14/CD16, CD27, and CD20 to discriminate various B cell subsets. B cells (AquaVital dye−, CD14−, CD16−, CD3−, CD20+) stained with both AlexaFluor 647 and Brilliant Violet 421-tagged HIV-1 CH505 T/F gp120 proteins or CH505 T/F stabilized SOSIP proteins were defined as gp120-specific or SOSIP-specific B cells. For the T cell subsets, a panel of markers including ICOS, CCR6, Bcl-6, CD183, CD69, CD185, Foxp3, Ki-67, CD3, PD-1, CD8a, CCR7, CD25, CD4 and CD45RA was used. Aqua Vital Dye was used to distinguish live from dead cells (Invitrogen).
Han Q., Bradley T., Williams W.B., Cain D.W., Montefiori D.C., Saunders K.O., Parks R.J., Edwards R.W., Ferrari G., Mueller O., Shen X., Wiehe K.J., Reed S., Fox C.B., Rountree W., Vandergrift N.A., Wang Y., Sutherland L.L., Santra S., Moody M.A., Permar S.R., Tomaras G.D., Lewis M.G., Van Rompay K.K, & Haynes B.F. (2020). Neonatal Rhesus Macaques Have Distinct Immune Cell Transcriptional Profiles following HIV Envelope Immunization. Cell reports, 30(5), 1553-1569.e6.
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3

Characterizing cervical tissue-resident memory T cells

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Cervical tissue from healthy donors was obtained and digested, and the resulting cell suspension was stained for viability with Aqua vital dye (Invitrogen) and the following antibodies: CD69-FiTC (FN50 dilution 1/20, cat. no. 557049), CD45-PE (HI30, dilution 1/100, cat. no. 560975), CD3-PerCP (SK7, dilution 1/10, cat. no. 345766), CD19-V500 (HIB19, dilution 1/50, cat. no. 561121), PD1-BV421 (EH12.1, dilution 1/20, cat. no. 562516) (all from BD Biosciences), CD4-APC (OKT4 BioLegend, dilution 1/80, cat. no. 300514). Aqua−CD19−CD45+ CD3+ CD4+ T cells were acquired in a BD FACSAriaTM II Cell Sorter and separated according to their CD69 expression into CD69+ (TRM) and CD69− (non-TRM). Sorted cells were then infected with 3655 TCID50 of the viral stock HIV-1BaL for 4 h at 37 °C or with medium in control conditions, washed with PBS and cultured in a 96-wells plate. After three days, cells were stained for viability with Live/Dead Fixable Far Red (Invitrogen), CD69-FiTC (FN50 dilution 1/20, cat. no. 557049) and HLA-DR-PerCP-Cy5.5 (G46-6, dilution 1/100, cat. no. 560652), before staining for p24 as described above. Fixed cells were finally acquired in a BD FACS Calibur flow cytometer and analyzed with FlowJo vX.0.7 software (TreeStar).
Cantero-Pérez J., Grau-Expósito J., Serra-Peinado C., Rosero D.A., Luque-Ballesteros L., Astorga-Gamaza A., Castellví J., Sanhueza T., Tapia G., Lloveras B., Fernández M.A., Prado J.G., Solé-Sedeno J.M., Tarrats A., Lecumberri C., Mañalich-Barrachina L., Centeno-Mediavilla C., Falcó V., Buzon M.J, & Genescà M. (2019). Resident memory T cells are a cellular reservoir for HIV in the cervical mucosa. Nature Communications, 10, 4739.
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4

Characterizing Macaque B and T Cell Subsets

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Macaque PBMCs were stained by a panel of markers, including CD38, Bcl-6, CD1c, CD69, CD21, Ki67, IgG, IgM, CD3/CD14/CD16, CD27, and CD20 to discriminate various B cell subsets. B cells (AquaVital dye−, CD14−, CD16−, CD3−, CD20+) stained with both AlexaFluor 647 and Brilliant Violet 421-tagged HIV-1 CH505 T/F gp120 proteins or CH505 T/F stabilized SOSIP proteins were defined as gp120-specific or SOSIP-specific B cells. For the T cell subsets, a panel of markers including ICOS, CCR6, Bcl-6, CD183, CD69, CD185, Foxp3, Ki-67, CD3, PD-1, CD8a, CCR7, CD25, CD4 and CD45RA was used. Aqua Vital Dye was used to distinguish live from dead cells (Invitrogen).
Han Q., Bradley T., Williams W.B., Cain D.W., Montefiori D.C., Saunders K.O., Parks R.J., Edwards R.W., Ferrari G., Mueller O., Shen X., Wiehe K.J., Reed S., Fox C.B., Rountree W., Vandergrift N.A., Wang Y., Sutherland L.L., Santra S., Moody M.A., Permar S.R., Tomaras G.D., Lewis M.G., Van Rompay K.K, & Haynes B.F. (2020). Neonatal Rhesus Macaques Have Distinct Immune Cell Transcriptional Profiles following HIV Envelope Immunization. Cell reports, 30(5), 1553-1569.e6.
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5

Multicolor Flow Cytometry Analysis of Immune Cells

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Blood samples (~500μl) were immediately lysed, washed, suspended in PBS and incubated with Aqua vital dye to distinguish live from dead cells (Invitrogen, Burlington, ON, Canada). Following two more washes, cells were suspended in washing and staining buffer (1% bovine serum albumin-PBS) and incubated for 20 minutes with the following cocktail of pre-titrated anti-mouse antibodies: CD3-Vioblue (145-2C11), CD4-APC-H7 (GK1.5), CD62L-PE (MEL-14-H2.100) (Miltenyi Biotec), CD44-Brilliant Violet 570 (IM7; BioLegend, San Diego, CA), CD11c-PE-Cy7 (HL3; BD Biosciences) and CCR5-FITC (CTC5), CXCR6-PerCP (221002) and CCR2-APC (475301) (R&D Systems Inc., Minneapolis, MN). All events were acquired in a BD FACSAria™ Cell Sorter and analyzed with FlowJo vX.0.7 software (TreeStar, Ashland, OR).
Qualai J., Cantero J., Li L.X., Carrascosa J.M., Cabré E., Dern O., Sumoy L., Requena G., McSorley S.J, & Genescà M. (2016). Adhesion Molecules Associated with Female Genital Tract Infection. PLoS ONE, 11(6), e0156605.
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