For example, the commonly used Hoechst 33342 stain, which binds to the minor groove of the double-stranded DNA can induce single-strand DNA breaks 5, or DRAQ5 (deep red fluorescing bisalkylaminoanthraquinone) the nuclear stain that intercalates with the cell’s DNA can influence chromation organization and lead to histone dissociation 6. Often these stains are incompatible with live cell analysis (for example, antibodies against histone modifications 3) and even if live cell reporters are available 4 these may have confounding effects on the cells. It can be achieved with conventional flow cytometry using multiple stains: typically, a stoichiometric fluorescent stain for DNA reports the cells’ position within the G1, S and G2 phases of the cell cycle 2, and additional stains are needed to sort mitotic cells into phases. For example, quantifying the proportion of cells in each phase of the cell cycle, including mitotic phases is very useful in the modern biological laboratory 3. The fluorescent stains can be used to label cellular components or processes, revealing specific cell phenotypes in the population and quantifying the particular state of each cell 2. It is highly suited to the study of cell populations and rare subset identification due to its high-throughput, multi-parameter nature. Conventional flow cytometry is a widespread and powerful technique for the measurement of cell phenotype and function using targeted fluorescent stains 1. A major challenge in many modern biological laboratories is obtaining information-rich measurements of cells in high-throughput and at single-cell resolution.
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