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Cornell University

Cornell Biotechnology Resource Center Flow Cytometry Facility

Technical Resources for BRC Flow Cytometry Facility Users

Tips for setting up proper controls in a multi-color experiment:

  1. Use a single color for each compensation control.
  2. A control tube must contain a negative and positive cell population to compare Mean Fluorescent intensities (MFIs).
  3. The negative and positive populations must have the same autofluorescence background.
  4. Use bright markers for setting compensation.
  5. The compensation control samples MUST have the same fluorochrome as the experimental sample.
  6. Tandem dyes have lot-specific compensation. Compensation must be redone if the user switches to a different manufacturing lot.
  7. Compensation beads can replace cells. Negative and positive beads MUST have the same autofluorescence. Compensation (capture) beads can be used with the same fluorochrome-conjugate lot used in the actual experiment.
  8. Resources on panel design: Fluorofinder, Panel Design Tips