We report the synthesis of a fluorescently end-labeled analog of a synthetic and degradable cationic poly(beta-amino ester) (PBAE; polymer 1) used in past studies for the delivery of DNA and the layer-by-layer assembly of erodible polyelectrolyte multilayers (PEMs). The synthesis of an analog of polymer 1 having acrylate-functionalized end groups provided a platform for the introduction of fluorescent labels by postpolymerization conjugate addition of amine-functionalized fluorophores. This approach enabled the synthesis of fluorescently end-labeled polymer (polymer 1(FL)) with molecular weights and polydispersities (M-n = 18,000; PDI similar to 1.8) similar to those used in past studies for the fabrication of PEMs using polymer 1. Layer-by-layer assembly of PEMs using polymer 1FL and poly(styrene sulfonate) enabled characterization of film erosion and, for the first time, direct observation of the release of cationic polymer from these assemblies using fluorescence microscopy and fluorometry. Our results shed new light on the behaviors of the cationic components of these PEMs and could prove useful for the design of thin films for a range of different controlled release applications. Our results also provide new fluorescent cationic polymer probes that could be useful for characterization of the behaviors of PBAEs in other fundamental or applied biotechnological contexts. (C) 2011 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 49: 1572-1581, 2011