The largest database of trusted experimental protocols

Live death vivid detection kit

Manufactured by Thermo Fisher Scientific
Sourced in United States, France

The Live/Death Vivid detection kit is a laboratory tool used to distinguish between live and dead cells. It provides a straightforward method for quickly assessing cell viability through fluorescent staining. The kit includes fluorescent dyes that selectively stain live and dead cells, enabling the user to visualize and quantify the population of each cell type.

Automatically generated - may contain errors

3 protocols using live death vivid detection kit

1

Multiparametric Flow Cytometry Analysis of PBMCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fresh PBMCs were isolated by density gradient centrifugation. FACS staining was carried out in PBS 1X supplemented with 2% FBS (foetal bovine serum) for 30 min at 4°C using the following fluorochrome-conjugated antibodies: APC-Vio770 anti-CD4, PerCP-Cy5.5 anti-CD3 (Milteniy Biotech, Paris, France), PE-CF594 anti-CD45RO, Alexa-Fluor 647 anti-CCR4, PE anti-CXCR3, PE-Cy7 anti-CCR6 (Becton Dickinson Biosciences, Pont de Claix, France). Intracellular staining was carried out using the FoxP3 permeabilization solution kit according to the manufacturer’s instructions (eBioscience, Paris, France) together with Alexa-Fluor 488 anti-SAMHD1 (from O. Schwartz’s lab; Institut Pasteur, Paris) and PE anti-Ki67 (Becton Dickinson). Dead cells were excluded using the Live/Death Vivid detection kit labeled with an aqua dye (Invitrogen). Fluorescence intensities were measured with an LSR II flow cytometer (Becton Dickinson) and analyzed using FlowJo version 10.0.7 (Tree Star Inc., Ashland, Oregon, USA).
+ Open protocol
+ Expand
2

Comprehensive B-cell Immunophenotyping by Flow Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols
Multi-color flow cytometry was performed on PBMC by acquiring 106 formaldehyde-fixed cells at the Gallios (Beckman Coulter) flow cytometer, and data analysed with FlowJo 8.8.7 (Treestar Inc.). The following mouse antihuman fluorochrome-conjugated monoclonal antibodies were used: CD19-PER-CP-Cy5.5 (SJ25-C1), CD10-PE-Cy7 (Hl10a), CD38-APC (HIT2), CD21-PE (B-ly4), CD27-V450 (M-T271), IgD-APC-H7 (IA6-2) and IgM-FITC (G20-127) (BD Biosciences). Dead cells were excluded using the Live/Death Vivid detection kit labeled with a near-infrared dye (Invitrogen, Carlsbad, California, USA). Gating strategy was performed as follows: only singlets were acquired and live B cells were gated on Vivid-CD19+ lymphocytes. Only samples with viability above 85% were subjected to the B-cell analysis. Transitional (“TR”) B cells were identified as CD19+CD27-CD10+. Thereafter, the CD10- cell population was gated on CD27++CD38high+ to identify Plasma-cells. Whereas on the CD27+CD38high- gate the following populations were identified: naive (“NV” CD27-CD21high+), resting memory (“RM” CD27+CD21high+), activated memory (“AM” CD27+CD21-) and tissue-like memory (“TLM” CD27-CD21-). RM, AM and TLM were each further characterized on the basis of Ig-expression into switched (“SW”, IgM-IgD-), Marginal Zone-like (“MZL”, IgM+IgD+), IgM-expressing (“IgM+”, IgM+IgD-) and IgD-expressing (“IgD+”, IgM-IgD+).
+ Open protocol
+ Expand
3

Multicolor Flow Cytometry of T-cell Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols
Multicolor flow cytometry was performed on fresh PBMCs and frozen LNMCs. T-cell subpopulations were determined using the different combinations of the following fluorochrome-conjugated antibodies: Pacific Blue, PE-Cy7 or Alexa-Fluor 700anti-CD3, PerCP or Alexa-Fluor 700 anti-CD4, PE anti-CD27, PE-Cy7 anti-CCR7, PE-CF594 anti-CD45RO, Alexa-Fluor 488 anti-CXCR5, Alexa-Fluor 647 anti-CCR4, PE anti-CXCR3, PE-Cy7 anti-CCR6 (Becton Dickinson Biosciences, Pont de Claix (Le), France). Vioblue anti-CD3, APC-Vio770 anti-CD4, APC-Vio770 anti-CD45RO, APC anti-CD28 (Milteniy Biotech, Paris, France). Brillant Violet 421 anti-CD279 (Biolegend, London, UK).
Intracellular staining was carried out using the FoxP3 permeabilization solution kit according to the manufacturer's instructions (eBioscience, Paris, France) together with PE Ki67, PE anti-Bcl-6 (BD Pharmingen) or PE anti-FoxP3 (Biolegend), and Alexa-Fluor 488 anti-SAMHD1 (a gift from O. Schwartz) or rabbit anti-SAMHD1 (Euromedex, Souffelweyersheim, France) followed by Alexa-Fluor 647 antirabbit antibody (Invitrogen, Life Technologies, Saint Auben, France).
Dead cells were excluded using the Live/Death Vivid detection kit labeled with an aqua dye (Invitrogen). Fluorescence intensities were measured with an LSR II flow cytometer (Becton Dickinson) and analyzed using FlowJo version 7.6.5 (Tree Star Inc., Ashland, Oregon, USA).
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!