The first demonstration of a molecularly imprinted polymer patterned by electron beam lithography (EBL) direct writing is reported. The polymeric mixture is based on a linear co-polymer that behaves simultaneously as a positive-tone EBL resist and, after polymerization in the presence of rhodamine 123 (R123) as a model analyte, as a selective and sensitive synthetic receptor for the template. Analyte binding was evaluated by fluorescence confocal microscopy and the imprinting effect was confirmed in the presence of compounds structurally related to R123.