Live dead fixable aqua stain fluorescence

Manufactured by Thermo Fisher Scientific

The LIVE/DEAD® Fixable Aqua stain is a fluorescent dye used to identify and distinguish between live and dead cells in flow cytometry applications. The dye binds to amine groups on the cell surface, emitting a fluorescent signal that can be detected using appropriate excitation and emission wavelengths.

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Lab products found in correlation

Lsr fortessa cytometer, by BD (2 mentions) Pe cd195, by BD (2 mentions) Cd4 alexafluor700, by BD (1 mentions) Cd8 apc h7, by BD (1 mentions) Cd69 pe cy7, by BD (1 mentions) Hla dr fitc, by BD (1 mentions) Fitc ki 67, by BD (1 mentions)

2 protocols using live dead fixable aqua stain fluorescence

Isolation and Phenotyping of Mucosal Mononuclear Cells

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Mucosal mononuclear cells (MMC) were isolated from rectal biopsies using a combination of mechanical and enzyme digestion ^{[26 ]}. Flow cytometric analysis was performed on a BD™ LSRFortessa cytometer (BD Biosciences, San Jose, CA). All antibodies were purchased from BD Biosciences, San Jose, CA (PerCP-CD45, Clone 2D1; Pacific Blue-CD3, Clone UCHT1; Alexa Fluor 700-CD4, Clone RPA-T4; APC-H7-CD8, Clone SK1; PE-Cy7-CD69, Clone FN50; FITC-HLA-DR, Clone L243; PE-CF594-CD38, Clone HIT2; FITC-Ki-67; APC-CD184 (CXCR4), Clone 12G5; and PE-CD195 (CCR5), Clone 2D7). Cells were stained with LIVE/DEAD® Fixable Aqua stain fluorescence (Life Technologies, Eugene, OR) to define viable cells.

McGOWAN I., WILKIN T., LANDOVITZ R.J., WU C., CHEN Y., MARZINKE M.A., HENDRIX C.W., RICHARDSON P., ESHLEMAN S.H., ANDRADE A., CHEGE W., ANDERSON P.L., McCAULEY M., FARLEY J., MAYER K.H., ANTON P., BRAND R.M., CRANSTON R.D, & GULICK R. (2019). The Pharmacokinetics, Pharmacodynamics, and Mucosal Responses to Maraviroc-Containing PrEP Regimens in Men who have Sex with Men. AIDS (London, England), 33(2), 237-246.

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Rectal Mucosal Mononuclear Cell Isolation

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Mucosal mononuclear cells (MMC) were isolated from rectal biopsies using a combination of mechanical and enzyme digestion as previously described [10 (link)]. Flow cytometric analysis was performed on a BD LSRFortessa cytometer (BD Biosciences, San Jose, CA). All data were stored in list mode and analyzed with BD FACSDIVA operating system and Flow Jo (Tree Star, Inc., Ashland, OR). All antibodies were purchased from BD Biosciences, San Jose, CA (PerCP-CD45, Clone 2D1; Pacific Blue-CD3, Clone UCHT1; PE-Cy7-CD4, Clone SK3; APC-H7-CD8, Clone SK1; FITC-CD69, Clone FN50; APC-CD184 (CXCR4), Clone 12G5 and PE-CD195 (CCR5)) and titrated under assay conditions to determine an optimum saturating dilution. Cells were stained with LIVE/DEAD Fixable Aqua stain fluorescence (Life Technologies, Eugene, OR) to define viable cells. The gating strategy used in this study can be found in S1 Fig.

Mcgowan I., Cranston R.D., Duffill K., Siegel A., Engstrom J.C., Nikiforov A., Jacobson C., Rehman K.K., Elliott J., Khanukhova E., Abebe K., Mauck C., Spiegel H.M., Dezzutti C.S., Rohan L.C., Marzinke M.A., Hiruy H., Hendrix C.W., Richardson-Harman N, & Anton P.A. (2015). A Phase 1 Randomized, Open Label, Rectal Safety, Acceptability, Pharmacokinetic, and Pharmacodynamic Study of Three Formulations of Tenofovir 1% Gel (the CHARM-01 Study). PLoS ONE, 10(5), e0125363.

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