Apr 08, 2025

Public workspaceFlow Cytometry

DOI
dx.doi.org/10.17504/protocols.io.kqdg3k5b7v25/v1
  • Lucas Blasco-Agell1,2,
  • Meritxell Pons-Espinal1,2,
  • Miquel Vila3
  • 1Department of Pathology and Experimental Therapeutics, Bellvitge University Hospital- IDIBELL, 08908 Hospitalet de Llobregat, Spain;
  • 2Institute of Biomedicine of the University of Barcelona (IBUB), Carrer Baldiri Reixac 15-21, Barcelona 08028, Spain.;
  • 3VHIR-CIBERNED-ASAP
  • Vilalab Public
Miquel Vila
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DOI: dx.doi.org/10.17504/protocols.io.kqdg3k5b7v25/v1
Protocol Citation: Lucas Blasco-Agell, Meritxell Pons-Espinal, Miquel Vila 2025. Flow Cytometry. protocols.io https://dx.doi.org/10.17504/protocols.io.kqdg3k5b7v25/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: April 01, 2025
Last Modified: April 08, 2025
Protocol Integer ID: 125892
Funders Acknowledgements:
ASAP
Grant ID: ASAP-020505
Abstract
Flow Cytometry
Cells preparation
Cells preparation
Cells are detached and centrifuged at 300xg for 10 minutes
Pellet is incubated with 1:50 conjugated antibody in MACS buffer for 15 minutes protected from the light
Cold MACS buffer is added
Cells are centrifuged at 300xg for 10 minutes
Cell pellet is resuspended in cold MACS buffer and incubated with 1:500 of Propidium iodide at RT for 5 minutes to determine cellular viability of the culture. Unstained cells are used as negative controls.
Staining and Fluorescence Analysis
Staining and Fluorescence Analysis
Analyzes of staining percentage and fluorescence intensities are done with Kaluza software (Beckman Coulter Life Sciences).