- Flow cytometry is a laser-based technology that allows quantitative single-cell analysis. It is used to analyze the characteristics of cells.
- Analysis of the cell cycle by DNA quantification can be done using a flow cytometry simulator. The DNA in cells can be stained by DNA binding dyes e.g. Propidium iodide.
- These dyes bind in proportion to the amount of DNA present in the cell.
- Cells in the S phase have more DNA than cells in the G1 phase. Cells in the G2 will have approximately twice the DNA content as cells in G1. Thus; they will take up proportionately more dye.
- The suspension of cells is then aspirated into a flow cell. Cells, surrounded by a narrow fluid stream, pass one by one through a focused laser beam. The light is either scattered or absorbed when it strikes a cell.
- Absorbed light of the appropriate wavelength is reemitted as fluorescence. This reflects the internal structure of the cell and its size and shape.
- Fluorescence scatter signals are detected, amplified, and analyzed by a series of photodiodes and a computer system.
- In the flow cytometry analysis, the end result is quantitative information about every cell analyzed. Large numbers of cells are analyzed in a short period of time (>1,000/sec). This gives the advantage of creating statistically valid information about cell populations.
The potential applications of flow cytometry technique include the detection and measurement of:
- Cell cycle: Reliable assessment of cells in G0/G1 phase versus S phase, G2, or polyploidy, including analysis of cell proliferation and activation.
- Cell viability/apoptosis.
- Identification and characterization of distinct subsets of cells within a heterogeneous sample.
- Protein expression.
- Protein post translational modifications.
- RNA, including IncRNA, miRNA, and mRNA transcripts.
