For cell cycle analysis, harvested cells were fixed in 1 mL of FBS-reduced cell culture medium and 10 mL of 70% ethanol (Roth, Karlsruhe, Germany) for at least 12 h at 4 °C. Thereafter, the cells were suspended in Ringer solution and stained for 60 min on ice with Hoechst 333258 (Invitrogen, Eugene, OR, USA) and examined by flow cytometry (Cytoflex, Beckman Coulter, Brea, CA, USA). The resulting data were analyzed by Kaluza Flow Cytometry Analysis 2.1 (Beckmann Coulter, Krefeld, Germany) showing the distribution of cells in the G0/G1, S and G2/M phases of the cell cycle.
Kaluza flow cytometry analysis 2
Manufactured by Beckman Coulter
Sourced in Germany
Kaluza Flow Cytometry Analysis 2.1 is a software suite designed for the analysis of flow cytometry data. It provides tools for data visualization, gating, and statistical analysis.
Automatically generated - may contain errors
2 protocols using kaluza flow cytometry analysis 2
1
Cell Cycle Analysis by Flow Cytometry
2
Quantifying Apoptosis and Necrosis by Flow Cytometry
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To detect apoptotic and necrotic cells using flow cytometry (Cytoflex, Beckman Coulter, Brea, CA, USA), harvested cells were suspended in Ringer solution and stained for 30 min on ice with Annexin V APC (BD, Heidelberg, Germany) and 7-amino-actinomycin D (7-AAD) (BD, Heidelberg, Germany). Kaluza Flow Cytometry Analysis 2.1 (Beckmann Coulter, Krefeld, Germany) was used to analyze the resulting data. Unstained cells (Annexin-negative/7-AAD-negative) were defined as viable cells. Annexin-positive/7-AAD-negative cells were defined as apoptotic and Annexin-positive/7-AAD-positive cells as necrotic (Figure 2 A).
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