Sample interview questions: How do you ensure the accuracy and reliability of immunofluorescence-based imaging flow cytometry experiments?
Sample answer:
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Use high-quality antibodies:
- Ensure antibody specificity by using antibodies validated for immunofluorescence-based imaging flow cytometry.
- Avoid cross-reactivity by selecting antibodies with minimal binding to non-target antigens.
- Store antibodies properly to maintain their activity and prevent degradation.
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Optimize staining conditions:
- Determine the optimal antibody concentration to minimize background staining and maximize signal-to-noise ratio.
- Optimize staining times and temperatures to ensure specific antibody binding and prevent non-specific interactions.
- Use appropriate buffers and blocking agents to reduce background staining and enhance signal-to-noise ratio.
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Use appropriate controls:
- Include unstained controls to determine the level of autofluorescence and non-specific staining.
- Use isotype controls to assess the specificity of antibody binding and exclude non-specific interactions.
- Include positive and negative controls to validate the staining protocol and ensure the accuracy of the results.
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Calibrate and maintain the flow cytometer:
- Regularly calibrate the flow cytometer to e… Read full answer