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Hoechst-IR is membrane impermeable: A–C): RAW 264.7 macrophage cells were incubated for 30 min with either Hoechst-IR (20 μM) or Hoechst 33258 (20 μM) and imaged by fluorescence microscopy, using an excitation of 340 nm. (A) Live cells treated with Hoechst-IR have negligible fluorescence, demonstrating that Hoechst-IR is membrane impermeable. (B) Fixed cells treated with Hoechst-IR and (C) live cells treated with Hoechst 33258 have high fluorescence.
