Date of Award

1999

Degree Name

Chemistry

College

College of Science

Type of Degree

M.S.

Document Type

Thesis

First Advisor

Michael Norton

Second Advisor

Robert Morgan

Third Advisor

William Price

Fourth Advisor

Leonard J. Deutsch

Abstract

Very few electron beam based methods for producing high-resolution patterns of surface bound proteins on glass have been developed.

There are only two types of end groups at the surface of glass, they are silanol and siloxane groups. An estersil layer can be formed on the surface of glass by treatment with hot alcohol creating a hydrophobic layer. The reaction is depicted as follows.

-Si-OH + HOR -SiO-R +H2O R = alkyl chain

The proposal that protein arrays could be fabricated by patterned electron bombardment using estersils as a negative resist was tested in this research. The hydrophobic estersil layer was expected to have a lower affinity to protein adhesion compared to the glass surface (silanol). Breakage of the estersil bond will expose the underlying glass surface for adhesion.

A JEOL JSM-5310LV scanning electron microscope (SEM) was used as the electron source for irradiation of coverslips. Laminin was the protein used for these patterning experiments. The rabbit polyclonal to laminin primary antibody and the goat anti-rabbit conjugated to fluorescein isothiocynate (FITC) secondary antibody were used to identify the localization of laminin. A Bio-Rad MRC-1024 confocal microscope was used to capture micrographs of the experiments.

Several experiments, performed using a nickel grid as a mask, supported this estersil resist hypothesis. However, when control experiments were conducted using an electron beam direct write (EBDW) maskless technique, the data disclosed a more complex mechanism of adsorption. Since the only variable removed in the EBDW experiments was the mask, it was possible to evaluate the contribution of the mask to the observed adhesion. It was concluded that sputtering of the nickel mask made a significant contribution to protein adsorption.

Two viable methods for high-resolution protein patterning using electron beam methods have been devised and investigated. Both methods warrant further development for use in cellular growth regulatory projects.

Subject(s)

Proteins – Research.

Biochemistry.

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