Photoacoustic molecular imaging, combined with the reporter-gene technique, can provide a valuable tool for cancer research. The expression of the lacZ reporter gene can be imaged using photoacoustic imaging following the injection of X-gal, a colorimetric assay for the lacZ-encoded enzyme β-galactosidase. Dual-wavelength photoacoustic microscopy was used to non-invasively image the detailed morphology of a lacZ-marked 9L gliosarcoma and its surrounding microvasculature simultaneously in vivo, with a superior resolution on the order of 10 μm. Tumor-feeding vessels were found, and the expression level of lacZ in tumor was estimated. With future development of new absorption-enhancing reporter-gene systems, we anticipate this strategy can lead to a better understanding of the role of tumor metabolism in cancer initiation, progression, and metastasis, and in its response to therapy.