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Issue 12, 2008
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Photolithographic patterning of organosilane monolayer for generating large area two-dimensional B lymphocyte arrays

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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.

Graphical abstract: Photolithographic patterning of organosilane monolayer for generating large area two-dimensional B lymphocyte arrays

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Article information


Submitted
18 Jun 2008
Accepted
25 Sep 2008
First published
24 Oct 2008

Lab Chip, 2008,8, 2105-2112
Article type
Paper

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

N. Li and C. Ho, Lab Chip, 2008, 8, 2105
DOI: 10.1039/B810329A

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