Microstamp patterns of biomolecules for high-resolution neuronal networks

• Published in: Medical & biological engineering & computing Vol. 36; no. 1; pp. 135 - 141
• Main Authors: BRANCH, D. W, COREY, J. M, WEYHENMEYER, J. A, BREWER, G. J, WHEELER, B. C
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer 1998; Springer Nature B.V
• Subjects: Aldehydes; Bioassay; Biological and medical sciences; Cell Biology - instrumentation; Cell structures and functions; Cells, Cultured; Fundamental and applied biological sciences. Psychology; Glass; Glass substrates; Humans; Miscellaneous; Molecular and cellular biology; Nerve Net; Neuroblastoma; Neurology; Proteins; Silicones; Tumor Cells, Cultured; Cell proliferation; Biomedical data processing; Cell culture; Patterning; High resolution; Surface properties; Lysine polymer; Neural network; Technique; Microstructure; In vitro; Tumor cell
• ISSN: 0140-0118; 1741-0444
• DOI: 10.1007/bf02522871

A microstamping technique has been developed for high-resolution patterning of proteins on glass substrates for the localisation of neurons and their axons and dendrites. The patterning process uses a microfabricated polydimethylsiloxane stamp with micrometer length features to transfer multiple types of biomolecules to silane-derivatised substrates, using glutaraldehyde as a homobifunctional linker. To test the efficacy 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 to verify the presence and uniformity of the patterns on the glass substrates. As a biological assay, B104 neuroblastoma cells were plated on stamped and physisorbed glass coverslips. Pattern compliance was determined as the percentage of cells on the pattern 8 h after plating. Results indicate that the stamping and photoresist patterning procedure are equivalent. Substrates stamped with PDL had an average pattern compliance of 52.6 +/- 4.4%, compared to 54.6 +/- 8.1% for physisorbed substrates. Measures of background avoidance were also equivalent. As the procedure permits successive stamping of multiple proteins, each with its own micropattern, it should be very useful for defining complex substrates to assist in cell patterning and other cell guidance studies.