Imaging β rays in vivo will help to advance microdosimetry and radiopharmaceutical development. In an earlier paper [1], we reported a newly developed system capable of directly imaging highenergy electron emissions in small animals in vivo. In this paper, we have thoroughly characterized the performance of the system. We have measured the sensitivity and detectability and the spatial resolution at various magnifications, as well as the linearity of the system. The system has also demonstrated the capability of directly detecting conversion electrons and positrons as well as β rays. The system has been applied to dynamically image spatiotemporal 18 F-Fluorodeoxyglucose (FDG) uptake distributions in xenograft small tumors in dorsal window chambers on mice in vivo. Heterogeneity in FDG uptake in millimeter-sized tumors has been observed.