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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
Lithium bis(trimethylsilyl)amide solution, 1.0 M in THF
Sigma-Aldrich
Hexamethyldisilazane, reagent grade, ≥99%
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Sodium bis(trimethylsilyl)amide solution, 1.0 M in THF
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Potassium bis(trimethylsilyl)amide solution, 1 M in THF
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Lithium bis(trimethylsilyl)amide, 97%
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Hexamethyldisilazane, ReagentPlus®, 99.9%
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Potassium bis(trimethylsilyl)amide, 95%
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Potassium bis(trimethylsilyl)amide solution, 0.5 M in toluene
Supelco
Hexamethyldisilazane, for GC derivatization, LiChropur, ≥99.0% (GC)
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Lithium bis(trimethylsilyl)amide solution, 1 M in toluene
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Sodium bis(trimethylsilyl)amide, 95%
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Lithium bis(trimethylsilyl)amide solution, 1.5 M in THF
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Lithium bis(trimethylsilyl)amide solution, 1.0 M in hexanes
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Sodium bis(trimethylsilyl)amide solution, 0.6 M in toluene
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Potassium bis(trimethylsilyl)amide solution, 1.0 M in 2-methyltetrahydrofuran
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Sodium bis(trimethylsilyl)amide solution, 0.5 M in methyl tert-butyl ether
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