Abstract We present a characterization methodology for fast direct measurement of the charge accumulated on Floating Gate (FG) transistors of Flash EEPROM cells. Using a Scanning Electron Microscope (SEM) in Passive Voltage Contrast (PVC) mode we were able to distinguish between '0' and '1' bit values stored in each memory cell. Moreover, it was possible to characterize the remaining charge on the FG; thus making this technique valuable for Failure Analysis applications for data retention measurements in Flash EEPROM. The technique is at least two orders of magnitude faster than state-of-the-art Scanning Probe Microscopy (SPM) methods. Only a relatively simple backside sample preparation is necessary for accessing the FG of memory transistors. The technique presented was successfully implemented on a 0.35 μm technology node microcontroller and a 0.21 μm smart card integrated circuit. We also show the ease of such technique to cover all cells of a memory (using intrinsic features of SEM) and to automate memory cells characterization using standard image processing technique.