Fluorescein diacetate

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  • fluorescein (494/518) is formed by intracellular hydrolysis of FDA
  • bacteria can hydrolyze this
  • leaks from cells
  • fluorescein has a relatively high rate of photobleaching
  • pH-sensitive fluorescence ref (pKa ~6.4) that is significantly reduced below pH 7
  • relatively broad fluorescence emission spectrum (not good for multicolor use)