Cell Cycle Analysis by Flow Cytometry
The cell cycle is a series of events that leads to the division and replication of the cell. Analysis of DNA content using a fluorescent nucleic acid stain enables quantification of the replication status of a cell.
Quiescent and G1 cells have one copy of DNA and therefore have a 1x fluorescence intensity. Cells in G2/M phase of the cell cycle have two copies of DNA and as such have a fluorescence intensity double that of G1 cells. Cells in S phase are synthesizing DNA and therefore have fluorescence values between those of the G1 and G2/M populations.
As many xenobiotics interfere with normal cell division, causing cells to arrest at specific points through the cell cycle, Gentronix Ltd offer the cell cycle assay to determine if a client’s compound is interfering with normal cell replication.
- Simple, robust and fast protocol
- Well established methodology
- p53-competent TK6 cell line
- Low compound requirement
Cells are exposed to several concentrations of test chemical for 24 hours. Following removal of the test chemical, the cells are lysed and fixed. Cells are then exposed to a fluorescent stain which binds to DNA. Single cell populations are then analysed for DNA content, which is directly proportional to the fluorescence of the nucleic acid stain.
Data are collected using BD FACS CantoTM ll and BD FACSCaliburTM flow cytometers, with a minimum of 10,000 cellular events scored per dose. Data are presented as percentages of the total cell population in each individual phase of the cell cycle. The study reports provide a detailed description of the study, data collection and results interpretation and are quality checked by a Gentronix expert flow cytometrist, independent to the study.