A versatile technique for patterning biomolecules onto glass coverslips

A fast, inexpensive, and versatile technique for patterning the surface of glass coverslips with molecules of biological interest is described. The technique combines photolithographic, silane-coupling, and protein adsorption procedures to pattern coverslips with amines, alkanes, and proteins with micrometer spatial resolution. The attachment of amines and alkanes was verified using contact angle and X-ray photoelectron spectroscopic (XPS) measurements. XPS results showed that amines and alkanes were attached in 1-4 nm thickness covering approximately 20% and 45%, respectively, of the surface. Patterns of amines were visualized using fluorescent staining, and patterns of proteins were detected immunochemically. Patterned coverslips were used to investigate adhesion and neurite outgrowth of mouse neuroblastoma (NIE-115) cells. Cells were examined on the following patterns: alkane-glass, protein-glass, amine-alkane, and amine-protein. Cell attachment and neurite outgrowth on patterned coverslips displayed the following preferences: laminin, fibronectin, or collagen IV > amine or glass > alkane or bovine serum albumin. This patterning method should be useful for studies of cell-surface interactions, cell migration, nerve regeneration, and the formation of neural networks in vitro.