Inhaled radioactive CO is currently the tracer of choice for blood volume quantification by positron emission tomography (PET). This measurement is of great interest for several clinical and research applications. However, owing to the short half-life of the radiolabeled CO, it can only be used in centers equipped with a cyclotron. In the present work, we propose an alternative method to label the red blood cells with [18F] in order to obtain blood volume measurements by PET. The use of the radioactive synthon [18F] N-succinimidyl 4-[18F]fluorobenzoate ([18F]SFB) was evaluated for erythrocyte labeling and PET blood volume imaging. The images provided by [18F]SFB labeled erythrocytes were compared with those obtained with inhaled [11C]CO. Blood volumes obtained with [18F]SFB labeled erythrocytes were similar to those obtained with [11C]CO in all of the evaluated organs with the exception of spleen, which presented lower uptake with this method. Since the [18F]-SFB binds irreversibly to red blood cells, in vivo stability of the radiolabel was higher compared with the [11C]CO method. Additionally, owing to the longer half-life and the shorter positron range of [18F], the image quality was also higher with the [18F]SFB radiolabeled erythrocytes. The labeling of red blood with [18F]SFB represents an advantageous alternative to radioactive CO for blood volume measurement by PET and cardiovascular isotopic imaging. Copyright © 2013 John Wiley & Sons, Ltd.