Cerenkov luminescence imaging of the PET radioisotope 68Ga in vitro. The positron-emitter 68Ga was eluted from a generator (Eckert & Ziegler, Berlin, Germany) and its activity was measured using a calibrated dose calibrator (Capintec CRC 212). The manufacturer specified setting and calibration factor for 68Ga was used for all measurements. Clear Eppendorf tubes containing various activity concentrations of 68Ga were placed within the imaging chamber of a commercial bioluminescence imaging system (IVIS 100; Perkin Elmer, Waltham, MA). The IVIS system consists of a cryogenically-cooled charged couple device (CCD) camera, operating at −90 °C, and a temperature controlled, light-tight imaging chamber. Two sets of images were collected; a white-light image and a luminescence image that captured photon output due to Cerenkov luminescence. Photon flux was quantified by means of proprietary software (Living Image version 2.5) using custom regions of interest based on the outlines of the Eppendorf tube caps visible on the white-light image. A. CLI image of 89 kBq of 68Ga. B. ROI analysis of photon flux demonstrated 1136,500 p/sec/cm2/sr. C. There is a positive linear relationship between the 68Ga activity and measured luminescence (R2 = 0.9996). The 68Ga activities ranged from 19 to 1689 kBq and were imaged in triplicate using the IVIS 100 high resolution (4) setting, FOV15, f1, open filter for 30 s (Courtesy of CLW, MVK and MFT).