Imaging of gene expression is increasingly essential for cancer diagnosis, prediction of tumor response to various available therapies, and for monitoring response to therapy, whether it is gene or nongene therapy. Most of the work on developing techniques and methodologies for imaging gene expression has been performed in mice. Sophisticated small animal imaging technologies have been developed for this purpose. Imaging of gene expression in humans is still very limited. Nuclear medicine, positron-emission tomography, magnetic resonance imaging, and optical methods are the modalities that have shown preliminary utility in imaging gene expression. Two major imaging strategies have been investigated: using marker genes encoding either intracellular enzymes or cell-surface receptors. The first approach exploits the ability of certain enzymes to modify imaging prodrugs, so that tissue accumulation of such drugs reflects the expression. The second approach uses cell-surface expression of ligand-binding receptors that can be detected using imaging tracers. In this review, we summarize some of the imaging techniques that have been developed to detect gene expression.