Modification of silicon nitride and silicon carbide surfaces for food and biosensor applications

Silicon-rich silicon nitride (SixN4, x > 3) is a robust insulating material widely used for the coating of microdevices: its high chemical and mechanical inertness make it a material of choice for the reinforcement of fragile microstructures (e.g. suspended microcantilevers, micro-fabricated membranes-“microsieves”) or for the coating of the exposed surfaces of sensors (field-effect transistors, waveguide optical detectors). To a more limited extent, silicon carbide (SiC) can find similar applications, and this material also starts to be more and more applied in coating and sensor technologies.
In all these applications, control over the surface properties of inorganic materials is crucial, for example to avoid blockage of membranes during filtration, or to provide sensor surfaces with specific (bio-)recognition properties. In this thesis, a variety of methods is developed to obtain and study robust functional coatings on SixN4 and SiC. These enable a whole new range of applications involving biocompatible and bio-specific surfaces, while retaining the bulk mechanical, structural, electrical or optical properties of the inorganic substrates.
Chapter 2 and 3 of the thesis give an overview of the great potential of covalent organic monolayers: Chapter 2 presents the formation of alkylthiol, alkylsilane and alkene monolayers, as well as a number of applications in biocompatible surfaces, micro- and nanopatterning of surfaces and sensing. The emphasis of this review chapter is put on the possible combinations of the bulk properties of inorganic materials (electrical, optical, structural) and the surface properties of organic monolayers (wettability, biospecificity, biorepellence). Chapter 3 is focused on biorepellent surfaces in the field of filtration with microfabricated membranes. Indeed, silicon nitride microsieves, despite their high permeability and structural homogeneity, are prone to pore blocking, when submitted to biological solutions. The chapter gives a review of the available surface modification techniques involving organic coatings that can minimize or even prevent this surface contamination. These coatings involve highly hydrophilic oligomers and polymers, which have been widely explored for organic surfaces. Covalent organic monolayers formed onto inorganic surfaces can extend the applications of these biorepellent coatings to microdevices like SixN4 microsieves (as also discussed in Chapters 7 and 8)
Chapter 4 and 5 present the thermal functionalization with highly stable alkene-based organic monolayers of the surfaces of silicon-rich silicon nitride (Chapter 4) and silicon carbide (Chapter 5). This work was motivated by the substantial knowledge of similar monolayer formation on silicon surfaces1,2 and the initial success of simple functionalizations on silicon nitride.3 The strong covalent attachment of the coating molecules with the substrates makes the obtained hybrid structures much more resistant to chemical degradation than other types of monolayers on these substrates. The reaction proceeds via attachment of the terminal double bond of alkenes with the surface groups (Si-H in the case of silicon nitride surfaces or –OH for silicon carbide surfaces). Besides methyl-terminated surfaces, functional coatings can be obtained by the use of bi-functionalized alkenes (Figure 1), also allowing further surface reactions and the attachment of bio-recognition elements, through covalent attachment of diverse chemical (carboxylic acid, amine) or biological (oligo-peptides, protein) moieties.

Figure 1. Modification of SiC and Si¬xN4 surfaces with alkyl monolayers
Chapter 6 describes a modification of this method, where UV irradiation is used instead of heat to initiate the modification of both silicon nitride and silicon carbide. For both materials, this method allows the grafting of heat-sensitive compounds, needs less starting material (using only a liquid film) and provides monolayers with higher quality (as e.g. indicated by grafting density and stability) and higher reproducibility. Here again the attachment of diverse functionalities is possible, via formation of activated esters. After hydrolysis and activation of such grafted ester, amines can be attached in high yield (> 80 %), as demonstrated using X-ray photoelectron spectroscopy (XPS). Besides the homogeneous modification of plain surfaces, this method also opens the way to surface patterning of silicon nitride and silicon carbide and the modification of mechanically sensitive microfabricated devices.
In Chapters 4 to 6, the chemical functionalizations are studied using X-ray photoelectron spectroscopy (XPS), infrared reflection absorption spectroscopy (IRRAS), atomic force microscopy (AFM), time-of-flight secondary ion mass spectrometry (ToF-SIMS) and static water contact angles. Si-C bonds are formed preferentially upon reaction of SixN4 surfaces with alkenes, similarly to what is reported for pure silicon surfaces, albeit that no measurement could totally exclude the presence of C-N bonds. The wet etching of SiC yields hydroxyl-terminated surfaces, and an IRRAS study reveals the attachment of alkenes via a Markovnikov-type addition (O-C bond formed on the second carbon of the double bond). The stability of these monolayers is reported in acidic and basic conditions, and it was shown that UV initiation yields even more stable monolayers, probably due to some cross-linking of the alkyl chains.
Chapter 7 explores the biorepellence of UV-initiated monolayers on silicon nitride surfaces Oligomers of ethylene glycols (3 or 6 units: methoxy-tri(ethylene oxide) undec-1-ene (CH3O(CH2CH2O)3(CH2)9CH=CH2; EO3, and methoxy-hexa(ethylene oxide) undec-1-ene (CH3O(CH2CH2O)6(CH2)9CH=CH2; EO6) are attached on the silicon nitride surfaces. The adsorption of two proteins, bovine serum albumin (BSA) and fibrinogen is used to test the biorepellence of the monolayers, in comparison with bare oxidized silicon nitride. Both proteins adsorb readily onto bare SixN4 surfaces, with adsorbed amounts of 1.25 and 2.7 mg.m-2 for BSA and fibrinogen, respectively, of which more than 80 % is irreversibly bound. In contrast to this, when oligomers are attached to the surface, this adsorption decreases to under the detection limit of the method used for this experiment (optical reflectometry). The ex situ study of surfaces with AFM and water contact angles also indicates that some of the monolayers completely prevent the adsorption of proteins.

Figure 2. Biorepellent behavior of oligoethylene oxide coated SixN4 surfaces
Chapter 8 describes the applications of the biorepellent coatings used in Chapter 7 (EO6) to silicon nitride microsieves, in order to improve the filtration of biological solutions and liquid food products. The EO6 coatings are successfully formed on microfabricated membranes with pore diameters of 0.45 micrometer, using the UV-initiated monolayer formation described in Chapter 6. This work shows that these coatings could be applied without loss of permeability due to wettability or pore blocking. Moreover, AFM showed that these coatings significantly decrease the adsorption of proteins on the surface between the pores.
Chapter 9 describes an alternative functionalization technique for inorganic surfaces, namely the use of plasma oxidation of alkyl monolayers to reproducibly form aldehydes (among other oxidized species) onto surfaces. The method described here for silicon and silicon nitride surfaces, is developed for the functionalization of sensitive devices and substrates. The formation of methyl-terminated alkyl monolayers from linear terminal alkenes is one of the easiest to perform, since linear monofunctional alkenes are readily available, their purification is easy (distillation) and their grafting conditions are very flexible (liquid state, heat-resistant, UV-resistant > 250 nm). Once these stable monolayers are formed, a short plasma treatment (0.5 to 2 s) is able to form oxidized functionalities within the top few angstroms of the surface, while the underlying alkyl chains retain their initial packing and insulation properties of the inorganic substrate. The grafting of gold nanoparticles shows that micron-sized patterns can be formed using a soft contact mask to protect a limited area of the monolayer. Alternatively, the aldehydes can be used to attach biotin and avidin onto SixN4 surfaces. The selective adsorption of biotinylated BSA onto the avidin-modified surfaces shows that the plasma treatment of methyl-terminated monolayers is a fast and efficient method to produce surfaces displaying high specific biochemical interactions.
In the chapter 10, some of the most striking effects that are described in the previous chapters are put into a wider perspective. Especially the formation and stability of monolayers is discussed, also in relation to biofunctionalization, biorepellence, and opportunities for surface engineering are proposed.