Sample interview questions: Can you explain the process of inducing apoptosis in cultured cells for specific experiments?
Sample answer:
1. Select a Suitable Cell Line:
- Choose a cell line that is susceptible to apoptosis induction.
- Confirm the presence of functional apoptotic pathways in the selected cell line.
2. Choose an Apoptosis-Inducing Agent:
- Consider a variety of agents known to induce apoptosis, including:
- Chemical inducers (e.g., staurosporine, etoposide, cisplatin)
- Natural compounds (e.g., curcumin, resveratrol, quercetin)
- Physical agents (e.g., heat shock, UV radiation, ionizing radiation)
3. Determine the Optimal Concentration and Duration of Treatment:
- Perform a dose-response experiment to determine the concentration of the apoptosis-inducing agent that results in the desired level of apoptosis.
- Optimize the duration of treatment to achieve the desired apoptotic response.
4. Monitor Apoptosis Induction:
- Assess apoptosis induction using various methods:
- Flow cytometry: Annexin V/propidium iodide staining to quantify apoptotic cells
- TUNEL assay: Detection of DNA fragmentation, a hallmark of apoptosis
- Caspase activity assays: Measurement of caspase activation, a key step in the apoptotic pathway
- Monitor cell viability using assays such as the MTT assay or trypan blue exclusion assay.
5. Collect and Analyze Data:
- Compile data on apoptotic cell death and cell viability.
- Analyze the results to determine the effectiveness of the apo… Read full answer
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