We report a new approach for forming ion depletion zones (IDZs) using intercalation reactions at an electrochemically controlled Prussian blue (PB) film. The experiments described here were performed using a microfluidic device, wherein a charged fluorophore in solution was used as a proxy to monitor IDZ formation. In the presence of K + , intercalation reactions proceed readily to form an IDZ, and thus enrich the fluorophore up to 37-fold. In the presence of the larger hydrated Li + or tetrabutylammonium (TBA +) ions, however, ion intercalation proceeded less quickly than with K +. Slower ion intercalation resulted in a weaker IDZ and correspondingly lower enrichment factors of eight-and six-fold, respectively. These results are significant because they show that PB intercalation reactions selectively form an IDZ. Accordingly, we anticipate that these findings will lead to a better understanding and further interesting applications of intercalation materials like PB for efficient and selective enrichment and separations.