The present study evaluated a new technique that has the potential to improve the border detection of in-stent neointima using an echogenic contrast agent during intravascular ultrasound (IVUS). To confirm the reproducibility of IVUS imaging for measuring the cross-sectional in-stent neointima area, inter- and intra-observer variability and correlation were determined. Conventional IVUS (plain IVUS) and IVUS using a contrast agent (contrast IVUS) were performed in 24 subjects 6.5+/-1.5 months after undergoing a Palmaz-Schatz coronary stent implant. Conventional IVUS delineated completely the in-stent neointima in 6 subjects (25%). In the remaining 18 subjects (75%), delineation of the neointima was incomplete despite the use of various combinations of imaging conditions (eg, transmission, compress, post-process). With contrast IVUS, the boundary of the neointima, and therefore the neointima area, was clearly distinguishable, and this resulted in complete delineation of the neointima in all 24 subjects. With a contrast agent, inter- and intra-observer variability significantly decreased (0.94+/-0.69mm2 conventional IVUS vs 0.37+/-0.40mm2 contrast IVUS, p<0.001; 0.69+/-0.56mm2 conventional IVUS vs 0.07+/-0.10mm2 contrast IVUS, p<0.0001; respectively). Thus, contrast IVUS provides a reproducible method for the quantitative analysis of in-stent neointima with excellent inter- and intra-observer correlation.