Nano-dot protein arrays are potent tools with broad applicability across diverse scientific disciplines such as biophysics, immunology, and healthcare. They present numerous advantages, particularly in facilitating investigations at the single cell or single molecule level. Among various nanofabrication methods, colloidal bead lithography (CBL) and electron beam lithography (EBL) are two complemetary choices that have been developped to produce metal-free protein nano-dot arrays that are compatible with all types of optical microscopy including super-resolution methods [1]–[4]. CBL stands out as an inexpensive choice [1]–[3] whereas EBL offers unparalleled flexibily and resolution [4]. Here we focus on EBL which we combin with surface functionalization to create protein nanodots within a passivated environment. These nanodots serve as valuable tool for studying cell adhesion. By systematically adjusting the size, shape, and spacing of the nanopatterns, we investigate how cells respond to distinct spatial constraints. Our approach achieves sub-40nm features, which are subsequently functionalized using two different strategies. The first strategy involves utilizing bioconjugated molecules, with biotine, while the second strategy leverages the contrast between PLL and PLL-PEG to precisely position the protein of interest within the nanodots. We demostrate the adhesion of various cell types on our nanopatterned substrates.
[1] doi: 10.1021/nl401696m.
[2] doi: 10.1021/acs.nanolett.5b01400.
[3] doi: 10.1021/acs.nanolett.7b01254.
[4] doi: 10.1021/acs.nanolett.1c01073