ResearchArticle In Situ Real-Time Chemiluminescence Imaging of Reactive Oxygen Species Formation from Cardiomyocytes

Correspondence should be addressed to Yunbo Li, yli@vcom.vt.edu and Ge Wang, wangg@vt.eduReceived 6 October 2008; Accepted 19 December 2008Recommended by Guowei WeiWe have applied the highly sensitive chemiluminescence (CL) imaging technique to investigate the in situ ROS formation incultured monolayers of rat H9c2 cardiomyocytes. Photon emission was detected via an innovative imaging system after incubationof H9c2 cells in culture with luminol and horseradish peroxidase (HRP), suggesting constitutive formation of ROS by thecardiomyocytes. Addition of benzo(a)pyrene-1,6-quinone (BPQ) to cultured H9c2 cells resulted in a 4-5-fold increase in theformation of ROS, as detected by the CL imaging. Both constitutive and BPQ-stimulated CL responses in cultured H9c2 cells weresustained for up to 1 hour. The CL responses were completely abolished in the presence of superoxide dismutase and catalase,suggesting the primary involvement of superoxide and hydrogen peroxide (H