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  • Photolithographic patterning of organosilane monolayer for generating large area two-dimensional B lymphocyte arrays.

Photolithographic patterning of organosilane monolayer for generating large area two-dimensional B lymphocyte arrays.

Lab on a chip (2008-11-22)
Nan Li, Chih-Ming Ho
ABSTRACT

High-density live cell array serves as a valuable tool for the development of high-throughput immunophenotyping systems and cell-based biosensors. In this paper, we have, for the first time, demonstrated a simple fabrication process to form the hexamethyldisilazane (HMDS) and poly(ethylene glycol) (PEG) binary molecular surface which can be used to effectively form high fidelity cell arrays. The HMDS self-assembled monolayer (SAM) on glass substrates was photolithographically patterned and its ability to physically adsorb proteins was characterized by contact angle measurement and fluorescence microscopy respectively. Passivation of the non-HMDS coated background by PEG was verified to have no impact on the pre-patterned HMDS and greatly inhibited the non-specific protein binding. Using the biotin-streptavidin complexation as an intermediate, uniform orientation and high bioactivity were achieved for the immobilized B lymphocyte specific anti-CD19 antibodies and therefore ensured the formation of high resolution B lymphocyte arrays. The cell-ligand interaction specificity was investigated and the anti-CD19 decorated micropatterns presented a much higher cell-capturing rate (88%) than those modified by non-specific ligands (15% for anti-CD5 and 7% for streptavidin). The approach was verified to be biocompatible and the properties of the antibody-modified surface were maintained after 12 h cell culture. The HMDS monolayer formation and patterning processes, and the universal HMDS/biotin-BSA/streptavidin template, provide a very simple and convenient process to generate high resolution micropatterns of cell-adhesive ligands and are extendable to form arrays of other types of cells as well.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Tris[N,N-bis(trimethylsilyl)amide]yttrium
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Tris[N,N-bis(trimethylsilyl)amide]samarium(III), 98%
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Hexamethyldisilazane, ReagentPlus®, 99.9%
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