Real-Time Fluorescence Measurements of ROS and [Ca²⁺] in Ischemic / Reperfused Rat Hearts: Detectable Increases Occur only after Mitochondrial Pore Opening and Are Attenuated by Ischemic Preconditioning - Fig 5

<p><b>ROS measurements using 5-cDCF surface fluorescence in control and IP hearts during ischemia / reperfusion A,</b> Typical traces for 5-cH₂DCFDA, diAM loaded control and IP hearts. <b>B,</b> Parallel data for hearts loaded with the ROS-insensitive dye, calcein-AM. The signal from a non-ischemic 5-cH₂DCFDA, diAM loaded heart (<b>A</b>, dark grey trace) demonstrates good dye retention during the time of the experiment. <b>C</b> and <b>D</b>, Mean data (± SEM, error bars) of 6 hearts loaded with 5-cH₂DCFDA, diAM (<b>C</b>) or calcein-AM (<b>D</b>). Signals were normalized using the mean fluorescence value obtained after 1 min of reperfusion. Note that loss of calcein signal with time is due to dye leakage and occurs even in normoxia. Statistically significant effects of IP are shown by the horizontal lines (** p ≤ 0.01; * p ≤ 0.05). Corresponding infarct sizes are presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0167300#pone.0167300.t001" target="_blank">Table 1</a>.