Cell Cycle Analysis
The cell cycle comprises G₀/G₁, S, and G₂/M phases, each defined by distinct DNA content: G₀/G₁ cells contain diploid DNA (2N), S-phase cells range from 2N to 4N, and G₂/M cells carry tetraploid DNA (4N). Using propidium iodide (PI) staining combined with flow cytometry, and after RNase digestion to remove RNA signals, we measure PI fluorescence intensity to quantify cellular DNA content. This allows precise discrimination of G₀/G₁, S, and G₂/M populations and enables software-driven calculation of each phase's percentage. Creative Diagnostics delivers accurate cell-cycle profiles to inform studies of proliferation dynamics, drug mechanisms, and gene-regulation effects.
Our Services
We offer a comprehensive PI-staining flow cytometry workflow for cell cycle analysis.
- Principle
PI intercalates into DNA; RNase treatment eliminates RNA staining. Flow cytometry measures PI fluorescence per cell, reflecting DNA content. The resulting histogram distinguishes G₀/G₁, S, and G₂/M phases. - Purpose
Quantify the distribution of cells across cycle phases to assess the impact of treatments or genetic modifications on cell-cycle progression.- Materials
- Cultured cell samples
- PI staining solution
- RNase digestion buffer
- PBS buffer
- Flow cytometry tubes and filters
- Procedure
- Collect and ethanol-fix cells from log-phase cultures.
- Treat with RNase to remove RNA interference.
- Stain with PI under dark conditions.
- Acquire fluorescence data by flow cytometry.
- Analyze with specialized software to fit DNA histograms and calculate phase percentages.
Key Advantages
With extensive flow-cytometry expertise and advanced instrumentation, we deliver high-sensitivity, highly reproducible cell-cycle analyses for any mammalian cell line or primary cell sample. Rigorous quality control at every step—from fixation and staining to data acquisition—ensures consistency and accuracy. We also customize synchronization or drug-treatment protocols to meet your specific research needs.
Workflow
Discuss your cell type, treatment conditions, and targeted phase metrics.
Define fixation, RNase digestion, and PI-staining parameters tailored to your cells.
Perform cell fixation, RNase treatment, PI staining, and flow-cytometry data collection.
Use professional software to model DNA-content distributions and compute phase percentages.
Deliver detailed histograms, phase-distribution charts, and written interpretation, plus recommendations for future experiments.
Notes
- Cell Quality: Ensure cells are in log-phase growth and free of contamination to avoid skewed results.
- Fixation: Thorough ethanol fixation prevents cell clumping and preserves DNA staining.
- Staining Conditions: Control PI concentration, incubation time, and light exposure to maintain signal integrity.
- Data Robustness: Acquire at least 10,000 events per sample for statistical reliability.
We look forward to providing precise, professional cell-cycle analysis to advance your research and drug-development projects!
