Feb 13, 2026
Cell apoptosis assay
- Guangli Suo1
- 1CAS Key Laboratory of Nano-Bio Interface, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Jiangsu, 215123, China.
- MTS
Protocol Citation: Guangli Suo 2026. Cell apoptosis assay. protocols.io https://dx.doi.org/10.17504/protocols.io.n92ld1jznl5b/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: February 13, 2026
Last Modified: February 13, 2026
Protocol Integer ID: 243183
Keywords: cell apoptosi, quantitative assessment of apoptotic cell population, apoptosi, apoptotic cell population, flow cytometry, using fluorescein isothiocyanate, fluorescein isothiocyanate, propidium iodide, assay
Abstract
Quantitative assessment of apoptotic cell populations by flow cytometry using fluorescein isothiocyanate (FITC)-conjugated annexin V and propidium iodide (PI) dual staining.
Troubleshooting
Cell apoptosis assay
The apoptosis of cells cultured on MWC-chips was assessed using Annexin V-FITC and propidium iodide (PI) double staining method (Vazyme). For cell lines, BRC cell lines (MDA-MB-231) or normal human foreskin fibroblasts (HFF) cell lines were seeded in the MWC-chips and cultured in Sel-Medium or normal cultural condition for 3 days. Subsequently, cells were harvested, trypsinized and stained with Annexin V-FITC and PI. For patient-derived cells, clinical BRC and adjacent tissues were dissociated into single cells which were then seeded onto MWC-chips for 2 days of culture in Sel-Medium followed by 3 days of culture in SG-Medium according to the MTSs generation protocol. The resulting MTSs and RCs were collected, trypsinized and stained with Annexin V-FITC and PI. Finally, the stained cells were analyzed by flow cytometry (CytoFLEX S, Coulter Beckman). Cells negative for PI but positive for Annexin V were considered apoptotic, while cells positive for both PI and Annexin V were classified as necrotic.