We propose a new technique for the analysis of micropatterned surfaces based on differences in local surface energy between different microdomains created on the surface by UV laser lithography, as a driving force for the selective immobilization of fluorescent biological probes of nanometric size. The method presented shows high detection resolution (in the order of the micrometer), requires far less specialized instrumentation, is easier and faster than conventional methods used today (autoradiographic techniques). The technique is thus a viable alternative to the use of radioactive probes, and may be used for the characterization of physical, chemical or topographical microheterogeneities on biological and non-biological surfaces. (C) 2003 Published by Elsevier B.V.