Fabrication and surface chemistry of nanoscale bioarrays designed for the study of cytoskeletal protein binding interactions and their effect on cell motility
(A) Schematic of the nanoarray chip. The chip contains an array of lattices with varying parameters , which defines the distance between protein binding sites. Each lattice has the same total number of binding sites of the same size. (B) Schematic of a strong binding event: ligand spacing matches the protein. (C) Schematic of a weak binding event: ligand spacing does not match the host protein.
Diagram of the Talin-Integrin Complex: The spacing of the integrin binding sites in the anti-parallel dimer of talin is about and a similar spacing is expected for the actin binding sites (adapted from Ref. 14).
Scanning electron micrographs of arrays of nanodot pairs patterned in HSQ. The interdot spacing ranges from to 80. The full range of dots pair arrays is part of a single chip. Inset: Magnified image of a single pair of dots. The dot diameter is .
Flow diagram for the functionalization of nanopatterned dot arrays.
Metal films patterned by direct evaporation through a metal shadow mask (i, k) Au and (j, l) Au (60%)/Pd (40%). (A) Untreated, (B) soaked for in fresh piranha, (C) soaked for in old piranha.
Atomic force microscope image of dots (A) before and (B) after PEGylation with HCl.
(A) Histograms of relative fluorescence intensities for surfaces PEGylated with 9, 18, acetic acid that were incubated with fluorescently labeled avidin. A histogram of instrument background is also plotted. (B) Topography of a surface PEGylated with acetic acid.
(A) Schematic and calculated heights of and . (B, C) Measured Ellipsometric thickness of thiol films vs the mole fraction of on freshly evaporated AuPd (B) and piranha treatedAuPd (C).
Dot arrays on glass are functionalized using fluorescent proteins as described in Fig. 5. (A) Shows localization of fluorescein labeled avidin, and (B) shows AlexaFluor 568 labeled fibronectin 7–10 domain derivative. The fluorescence from the PEGylated background is negligible.
Cells seeded on dot arrays such as shown in (D), where the dots are functionalized with a pentamer of the fibronectin 7–10 domain. The diameters of single dots are varied between 15 and , while the distance between them L is varied from such as to keep the total area covered by the dots constant at . The rest of the glass slide is passivated with a PEG-silane. Cells spread on dot arrays are shown after (A) , (B) , (C) , after seeding. No cell adhesion to the PEG passivated area is observed. Cells do not detach from continuous, functionalized metal films.
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