A novel optical diffraction based technique is proposed and demonstrated to measure deflections of the order of ~1nm in microcantilevers (MC) designed for sensing ultra-small forces of stress. The proposed method employs a double MC structure where one of the cantilevers acts as the active sensor beam, while the other as a reference. The active beam can respond to any minute change of stress, for example, molecular recognition induced surface stress, through bending (~1nm) relative to the other fixed beam. Optical diffraction patterns obtained from this double slit aperture mask with varying slit width, which is for the bending of MC due to loading, carries the deflection profile of the active beam. A significant part of the present work explores the possibility of connecting diffraction minima (or maxima) to the bending profile of the MC structure and thus the possibility to measure induced surface stress. To start with, it is also the aim to develop double MC sensors using PHDDA (Poly – Hexane diol diacrylate) because this material has the potential to achieve high mechanical deformation sensitivity in even moderately scaled down structures by virtue of its very low Young’s modulus. Moreover, the high thermal stability of PHDDA also ensures low thermally induced noise floors in microcantilever sensors. To demonstrate the proposed optical diffraction-based profiling technique, a bent microcantilever structure is designed and fabricated by an in-house developed Microstereolithography (MSL) system where, essentially one of the microcantilevers is fabricated with a bent profile by varying the gap between the two structures at each cured 2D patterned layer. The diffraction pattern obtained on transilluminating the fabricated structure by a spherical wavefront is analyzed and the possibility of obtaining the deflections at each cross section is ascertained. Since the proposed profiling technique relies on the accurate detection and measurement of shifts of intensity minima on the image plane, analysis of the minimum detectable shift in intensity minima for the employed optical interrogation setup with respect to the minimum detectable contrast and SNR of the optical measurement system is carried out, in order to justify the applicability of the proposed minima intensity shift measurement technique. The proposed novel diffraction based profiling technique can provide vital clue on the origins of surface stress at the atomic and molecular level by virtue of the entire bent profile due to adsorption induced bending thereby establishing microcantilever sensor technology as a more reliable and competitive approach for sensing ultra-low concentrations of biological and chemical agents.

This thesis uses surface sensitive tools to characterize the effect of a solid surface on immobilized biomacromolecules. This includes understanding how the surface can change the affinity of these macromolecules to small molecules compared to bulk studies. Two classes of immobilized biomacromolecules, the supported lipid bilayer (SLB) and the Lac repressor protein (LacI), are characterized using microcantilever sensors and quartz crystal microbalance with dissipation (QCM-D). The first part of this thesis reports the use of microcantilever beams, an ultrasensitive sensor for measuring the surface free energy changes on a substrate induced by molecular adsorptions, to probe the interaction between a solid surface and a phospholipid bilayer. This sensing method integrates two well-developed techniques: solid-supported lipid bilayers (SLBs) and the microcantilever (MC) sensors. Studying the adsorption free energy of lipid bilayers on a solid surface allows better characterizing of the formation and stability of SLBs. Microcantilever converts the Gibbs free energy change taking place on its surface into a mechanical deformation. As molecules physisorb or chemisorb onto the surface of the microcantilevers, the microcantilevers bend, either due to induced compressive or tensile stresses, which result from the surface free energy change. By monitoring the deflection values of the microcantilevers, the real-time surface free energy change during the SLB formation can be detected. This thesis has led to the development of a novel biosensor--lipid membrane coated microcantilevers--to detect the adsorption, insertion, aggregation and solubilizing effect of membrane-active substances, such as surfactants and peptides, on the phospholipid membranes. To better characterize the surface free energy, SLBs doped with charged lipids or cholesterol are shown to alter the surface free energy. We can predict this change in surface free energy using a thermodynamic model. Application of this membrane-coated cantilever is put into use for detecting how amphiphilic molecules interact with SLBs, as well as for probing the abrupt conformational change of SLBs during a temperature induced phase-transition. This study systematically demonstrates various usage aspects of microcantilever to characterize the SLBs, and how this technique may advance the biophysical knowledge of the lipid membrane, one of the essential building blocks of life. The second part of this thesis reports the use of both microcantilever sensors and QCM-D to measure the adsorption free energy and mass of a model protein, the Lac repressor (LacI), and compare how a modified T334C mutant that includes a cysteine group to orient the protein on the gold surface through a covalent sulfur bonds retains its binding capabilities over that of wild type LacI. The main challenge of this work is to unravel how the adsorption of biomacromolecules at the solid/liquid interface leads to surface free energy changes and ultimately changes the stress of the underlying solid surface (the cantilever). The uses of microcantilever sensors and QCM to probe the interactions that take place on SLBs and surface-bound proteins have the advantage of being a sensitive, real-time, and label-free technique.

The work elucidates the novel physical and thermal properties of thin and ultra-thin films of crosslinked polymer and organized microstructures with a special emphasis on surface and interfacial effects and the structure-property relationships. Two major crosslinked polymer coatings have been thoroughly investigated: polymer microstructures fabricated by multi-laser interference lithography (IL), and plasma polymer coatings. We unveiled intriguing thermal properties of plasma polymer films originating from their physical state and exploiting the same for the design of ultrasensitve chemical sensors. A novel paradigm of surface coatings, single and bi-component periodic, porous crosslinked polymeric structures, has been introduced and thoroughly studied. Surface, interfacial, and mechanical properties of these novel class crosslinked polymer coatings clearly demonstrate the enormous potential of the IL microstructures as organized multicomponent polymer systems. When subjected to external or internal stresses the periodic porous structures can exhibit a sudden and dramatic pattern transformation resulting in remarkable change in the photonic, phononic and mechanical properties of these structures. Furthermore, the confinement of these instabilities to localized regions results in complex hierarchical structures. The two polymer coatings (plasma polymers and IL microstructures) with complementary attributes (such as periodic structure, vertical stratification, residual internal stresses, and high surface and interface tunability) enabled us to understand and design novel multifunctional polymer coatings.