Surface-stress-based microcantilever aptasensor

This thesis presents the design, modelling, fabrication, and biological evaluation of a microcantilever-based aptasensor. It is the first reported work on aptasensors with aptamer immobilized on a bare SU-8 surface. Aptasensor surface funtionalisation was achieved using gas plasma treatment. Label-free detection of thrombin molecules using the aptasensor was successfully demonstrated.

A surface-stress-based microcantilever aptasensor

Biosensors based on microcantilevers convert biological recognition events into measurable mechanical displacements. They offer advantages such as small size, low sample volume, label-free detection, ease of integration, high-throughput analysis, and low development cost. The design and development of a microcantilever-based aptasensor employing SU-8 polymer as the fabrication material is presented in this paper. Aptamers are employed as bioreceptor elements because they exhibit superior specificity compared to antibodies due to their small size and physicochemical stability. To immobilise thrombin DNA aptamer on the bare SU-8 surface of the aptasensor, a combined plasma mode treatment method is implemented which modifies the surface of the aptasensor. Label-free detection of thrombin molecules using the fabricated aptasensor is successfully demonstrated. The measured deflection is one order of magnitude higher than that of a silicon nitride microcantilever biosensor. The developed aptasensor also demonstrates high specificity.

A surface-stress-based microcantilever aptasensor

Biosensors based on microcantilevers convert biological recognition events into measurable mechanical displacements. They offer advantages such as small size, low sample volume, label-free detection, ease of integration, high-throughput analysis, and low development cost. The design and development of a microcantilever-based aptasensor employing SU-8 polymer as the fabrication material is presented in this paper. Aptamers are employed as bioreceptor elements because they exhibit superior specificity compared to antibodies due to their small size and physicochemical stability. To immobilise thrombin DNA aptamer on the bare SU-8 surface of the aptasensor, a combined plasma mode treatment method is implemented which modifies the surface of the aptasensor. Label-free detection of thrombin molecules using the fabricated aptasensor is successfully demonstrated. The measured deflection is one order of magnitude higher than that of a silicon nitride microcantilever biosensor. The developed aptasensor also demonstrates high specificity.

Theoretical modeling and experimental validation of surface stress in thrombin aptasensor

Adsorption of target molecules on the immobilized microcantilever surface produced beam displacement due to the differential surface stress generated between the immobilized and non-immobilized surface. Surface stress is caused by the intermolecular forces between the molecules. Van der Waals, electrostatic forces, hydrogen bonding, hydrophobic effect and steric hindrance are some of the intermolecular forces involved. A theoretical framework describing the adsorption-induced microcantilever displacement is derived in this paper. Experimental displacement of thrombin aptamer-thrombin interactions was carried out. The relation between the electrostatic interactions involved between adsorbates (thrombin) as well as adsorbates and substrates (thrombin aptamer) and the microcantilever beam displacement utilizing the proposed mathematical model was quantified and compared to the experimental value. This exercise is important to aid the designers in microcantilever sensing performance optimization.

Effects of design parameters on sensitivity of microcantilever biosensors

Microcantilever biosensors produce cantilever bending due to differential surface stress between upper and lower surfaces of the cantilever. The bending is associated with concentration of ligands and adsorbed ligand-receptor intermolecular forces. Sample volume sizes in clinical diagnostic applications are usually very minute requiring a highly sensitive microcantilever for disease detection. This paper investigates a number of parameters that influence the sensitivity of microcantilever biosensors. The parameters include length, thickness, shape, and material of the cantilever beam. Biosensors of varying parameters are modeled and simulated. The results show that increasing the length of the cantilever beam enhances its sensitivity. However, increasing the thickness of the cantilever beam reduces its sensitivity. In static analysis, the shape of the cantilever beam does not notably impact upon its sensitivity. Also, using materials with lower Young’s modulus improves the sensitivity.

Design and analysis of a cantilever biosensor based on a boron nitride nanotube

In this paper, we introduce a single-walled boron nitride nanotube (SWBNNT)-based cantilever biosensor, and investigate its bending deformation. The BNNT-based cantilever is modelled by accounting that the surface of the cantilever beam is coated with the antibody molecule. We have considered two main approaches for the mechanical deformation of the BNNT beam. The first one is differential surface stress produced by the binding of biomolecules onto its surface, and the second one is the charge released from the biomolecular interaction. In addition, other parameters including length of beam, variation of beam’s location and chiralities of the BNNT have been taken into consideration to design the cantilever biosensor. The computed results are in good agreement with the well known electrostatic equations that govern the deformation of the cantilever.

Modeling and comparative analysis of carbon nanotube and boron nitride nanotube cantilever biosensors

This paper investigates the bending deformation of a cantilever biosensor based on a single-walled carbon nanotube (CNT) and single-walled boron nitride nanotube (BNNT) due to bioparticle detection. Through 3-D modeling and simulations, the performance of the CNT and BNNT cantilever biosensors is analyzed. It is found that the BNNT cantilever has better response and sensitivity compared to the CNT counterpart. Additionally, an algorithm for an electrostatic-mechanical coupled system is developed. The cantilever (both BNNT and CNT) is modelled by accounting that a conductive polymer is deposited onto the nanotube surfaces. Two main approaches are considered for the mechanical deformation of the nanotube beam. The first one is differential surface stress produced by the binding of biomolecules onto the surface. The second one is the charge released from the biomolecular interaction. Also, different ambient conditions are considered in the study of sensitivity. Sodium Dodisyl Sulphate (SDS) provides better bending deformation than the air medium. Other parameters including length of beam, variation of beam's location, and chiralities are considered in the design. The results are in excellent agreement with the electrostatic equations that govern the deformation of cantilever.

A micromechanical biosensor with interdigitated capacitor readout

The growing demands of high-throughput, accurate and fast response biological or chemical sensors are driving the development of new detection technologies. This paper presents a micromechanical biosensor with capacitive read-out method. The proposed biosensor design consists of a fixed-fixed beam attached to an interdigitated capacitor. Implementation of the interdigitated capacitor design improves the sensitivity of the biosensor. The effects of the electrode thickness, length and the number of electrode fingers on the change of capacitance are investigated. The results show that the percentage change of capacitance is proportional to the number of the electrode fingers. Similarly, the increase in the length of the electrodes results in an increase in the percentage change of the capacitance. However, as the thickness of the electrode increases, the percentage change of the capacitance decreases.

Effect of membrane stress on surface roughness changes in sheet forming

Friction plays an important role in sheet metal forming (SMF) and the roughness of the surface of the sheet is a major factor that influences friction. In finite element method (FEM) models of metal forming, the roughness has usually been assumed to be constant; even though it is commonly observed that sheet drawn under tension over a tool radius results in the surface becoming shiny, indicating a major change in surface morphology. An elastic–plastic FEM model for micro-contact between a flat surface and a single roughness peak has been developed. The model was used to investigate the effect of the membrane stress in the sheet on the deformation of an artificial roughness peak. From the simulation results, the change in asperity, or deformation of the local peak, for a given nominal tool contact stress is significantly influenced by the local substrate stress. The height of the asperity decreases with increasing substrate stress and the local pressure is much higher than the nominal pressure. In addition, the local contact stress decreases with an increase in the substrate stress levels.

Cement composites reinforced with surface modified coir fibers

An experimental investigation of coir mesh reinforced mortar (CMRM) is conducted using nonwoven coir mesh matting. The main parameters in this study are the fiber volume fraction (number of mesh layers) and fiber surface treatment with a wetting agent. The composites are subjected to the four-point bending test. The short-term mechanical properties of CMRM are discussed. Scanning electron micrograph analysis is used to observe the fiber—matrix interfacial characteristics. The results indicate that the addition of coir mesh to mortar significantly improves the composite post-cracking flexural stress, toughness, ductility, and toughness index, compared to plain mortar materials. The Albatex © FFC wetting agent (2-ethylhexanol) can effectively improve water absorption of coir fiber and enhance the fiber—matrix bonding strength. These coir mesh reinforced composites may be useful in civil engineering applications.

Heterologous microarray experiments used to identify the early gene response to heat stress in a coral reef fish

Coral reef fishes are expected to experience rising sea surface temperatures due to climate change. How well tropical reef fishes will respond to these increased temperatures and which genes are important in the response to elevated temperatures is not known. Microarray technology provides a powerful tool for gene discovery studies, but the development of microarrays for individual species can be expensive and time-consuming. In this study, we tested the suitability of a Danio rerio oligonucleotide microarray for application in a species with few genomic resources, the coral reef fish Pomacentrus moluccensis. Results from a comparative genomic hybridization experiment and direct sequence comparisons indicate that for most genes there is considerable sequence similarity between the two species, suggesting that the D. rerio array is useful for genomic studies of P. moluccensis. We employed this heterologous microarray approach to characterize the early transcriptional response to heat stress in P. moluccensis. A total of 111 gene loci, many of which are involved in protein processing, transcription, and cell growth, showed significant changes in transcript abundance following exposure to elevated temperatures. Changes in transcript abundance were validated for a selection of candidate genes using quantitative real-time polymerase chain reaction. This study demonstrates that heterologous microarrays can be successfully employed to study species for which specific microarrays have not yet been developed, and so have the potential to greatly enhance the utility of microarray technology to the field of environmental and functional genomics.

Mechanical properties and bioactive surface modification via alkali-heat treatment of a porous ti-18Nb-4Sn alloy for biomedical applications

A porous Ti–18 at.%Nb–4 at.%Sn (hereafter, Ti–18Nb–4Sn) alloy was prepared by powder metallurgy. The porous structures were examined by scanning electron microscopy and the phase constituents were analysed by X-ray diffraction. Mechanical properties of the porous alloy were investigated using a compressive test. To enhance the bioactivity of the alloy surface, alkali-heat treatment was used to modify the surface. The bioactivity of the pre-treated alloy sample was investigated using a biomimetic process by soaking the sample into simulated body fluid (SBF). Results indicate that the elastic modulus and plateau stress of the porous Ti–18Nb–4Sn alloy decrease with decreasing relative density. The mechanical properties of the porous alloy can be tailored to match those of human bone. After soaking in SBF for 7 days, a hydroxyapatite layer formed on the surface of the pre-treated porous Ti–18Nb–4Sn alloy. The pre-treated porous Ti–18Nb–4Sn alloy therefore has the potential to be a bioactive implant material.

Surface behaviour and lipid interaction of Alzheimer beta-amyloid peptide 1-42: a membrane-disrupting peptide

Amyloid aggregates, found in patients that suffer from Alzheimer's disease, are composed of fibril-forming peptides in a β-sheet conformation. One of the most abundant components in amyloid aggregates is the β-amyloid peptide 1–42 (Aβ 1–42). Membrane alterations may proceed to cell death by either an oxidative stress mechanism, caused by the peptide and synergized by transition metal ions, or through formation of ion channels by peptide interfacial self-aggregation. Here we demonstrate that Langmuir films of Aβ 1–42, either in pure form or mixed with lipids, develop stable monomolecular arrays with a high surface stability. By using micropipette aspiration technique and confocal microscopy we show that Aβ 1–42 induces a strong membrane destabilization in giant unilamellar vesicles composed of palmitoyloleoyl-phosphatidylcholine, sphingomyelin, and cholesterol, lowering the critical tension of vesicle rupture. Additionally, Aβ 1–42 triggers the induction of a sequential leakage of low- and high-molecular-weight markers trapped inside the giant unilamellar vesicles, but preserving the vesicle shape. Consequently, the Aβ 1–42 sequence confers particular molecular properties to the peptide that, in turn, influence supramolecular properties associated to membranes that may result in toxicity, including: 1), an ability of the peptide to strongly associate with the membrane; 2), a reduction of lateral membrane cohesive forces; and 3), a capacity to break the transbilayer gradient and puncture sealed vesicles.