The measurement of regional cerebral blood flow (rCBF) by single-photon emission computed tomography (SPECT) is a powerful clinical and research tool. There are several clinical applications now documented, a substantial number under active investigation, and a larger number yet to be studied. Standards regarding patient imaging environment and image presentation are becoming established. This article reviews key aspects of SPECT functional brain imaging in clinical practice, with a particular emphasis on therapeutics, including 1) the quality of the tomographic device, 2) the radiopharmaceutical employed, 3) environmental conditions at the time of radiotracer administration, 4) characteristics of the subject, 5) the format used for image presentation, and 6) the essential components of image processing necessary to the achievement of high-quality SPECT brain images. Next, a brief description of relevant radiation safety issues is provided. Finally, applications in molecular imaging, especially in small animal imaging for research as well as drug discovery and development are discussed. The gamut of SPECT studies from currently routine clinical applications to molecular imaging offers a wonderful frontier for opportunities to employ functional brain imaging in neurotherapeutics.