Calibration of the spring constant of cantilevers of arbitrary shape using the phase signal in an atomic force microscope.

The measurement of cantilever parameters is an essential part of performing a calibrated measurement with an atomic force microscope (AFM). The thermal motion method is a widely used technique for calibrating the spring constant of an AFM cantilever, which can be applied to non-rectangular cantilevers. Given the trend towards high frequency scanning, calibration of non-rectangular cantilevers is of increasing importance. This paper presents two results relevant to cantilever calibration via the thermal motion method. We demonstrate the possibility of using the AFM's phase signal to acquire the thermal motion. This avoids the challenges associated with connecting the raw photodiode signal to a separate spectrum analyser. We also describe how numerical calculations may be used to calculate the parameters needed in a thermal motion calibration of a non-rectangular cantilever. Only accurate knowledge of the relative size of the in-plane dimensions of the cantilever is needed in this computation. We use this pair of results in the calibration of a variety of rectangular and non-rectangular cantilevers. We observe an average difference between the Sader and thermal motion values of cantilever stiffness of 10%.

Microfabricated tissue gauges to measure and manipulate forces from 3D microtissues

Physical forces generated by cells drive morphologic changes during development and can feedback to regulate cellular phenotypes. Because these phenomena typically occur within a 3-dimensional (3D) matrix in vivo, we used microelectromechanical systems (MEMS) technology to generate arrays of microtissues consisting of cells encapsulated within 3D micropatterned matrices. Microcantilevers were used to simultaneously constrain the remodeling of a collagen gel and to report forces generated during this process. By concurrently measuring forces and observing matrix remodeling at cellular length scales, we report an initial correlation and later decoupling between cellular contractile forces and changes in tissue morphology. Independently varying the mechanical stiffness of the cantilevers and collagen matrix revealed that cellular forces increased with boundary or matrix rigidity whereas levels of cytoskeletal and extracellular matrix (ECM) proteins correlated with levels of mechanical stress. By mapping these relationships between cellular and matrix mechanics, cellular forces, and protein expression onto a bio-chemo-mechanical model of microtissue contractility, we demonstrate how intratissue gradients of mechanical stress can emerge from collective cellular contractility and finally, how such gradients can be used to engineer protein composition and organization within a 3D tissue. Together, these findings highlight a complex and dynamic relationship between cellular forces, ECM remodeling, and cellular phenotype and describe a system to study and apply this relationship within engineered 3D microtissues.

SU8 bio-chemical sensor microarrays

Microfabricated cantilevers have recently attracted considerable attention as novel label-free chemical and biological biosensors which translate surface reactions into nanomechanical bending motion. However these studies have primarily focused on commercially available silicon cantilevers and relatively little work has been performed on cantilevers fabricated from other materials. Polymeric materials, offer significant advantages over silicon by virtue of the low Young's modulus, ease of microfabrication and reduced cost. In this paper, we report a non-vacuum fabrication process to produce arrays of SU8 cantilevers and demonstrate their application as chemical sensors using in situ reference cantilevers. © 2006 Elsevier B.V. All rights reserved.