MICROSTAMP PATTERNS OF BIOMOLECULES FOR HIGH-RESOLUTION NEURONAL NETWORKS

A microstamping technique has been developed for high-resolution patte rning of proteins on glass substrates for the localisation of neurons and their axons and dendrites. The patterning process uses a microfabr icated polydimethylsiloxane stamp with micrometer length features to t ransfer multiple types of biomolecules to silane-derivatised substrate s, using glutaraldehyde as a homobifunctional linker. To test the effi cacy of the procedure, substrates are compared in which poly-d-lysine (PDL) was physisorbed and patterned by photoresist with those stamped with PDL. Fluorescein isothiocyanate labelled poly-l-lysine was used t o verify the presence and uniformity of the patterns on the glass subs trates. As a biological assay, B104 neuroblastoma cells were plated on stamped and physisorbed glass coverslips. Pattern compliance was dete rmined as the percentage of cells on the pattern 8h after plating. Res ults indicate that the stamping and photoresist patterning procedure a re equivalent. Substrates stamped with PDL had an average pattern comp liance of 52.6+/-4.4%, compared to 54.6+/-8.1% for physisorbed substra tes. Measures of background avoidance were also equivalent. As the pro cedure permits successive stamping of multiple proteins, each with its own micropattern, it should be very useful for defining complex subst rates to assist in cell patterning and other cell guidance studies.