Investigation of biotin-streptavidin binding interactions using microcantilever sensors

We report the investigation of biotin-streptavidin binding interactions using microcantilever sensors. A symmetric cantilever construction is employed to minimize the effects of thermal drift and the control of surface chemistry on the backside of the cantilever is demonstrated to reduce the effects of non-specific binding interactions on the cantilever. Three structurally different biotin modified cantilever surfaces are used as a model system to study the binding interaction with streptavidin. The cantilever response to the binding of streptavidin on these biotin sensing monolayers is compared. The lowest detection limit of streptavidin using biotin-HPDP is found to be between 1 and 10 nM limited by the optical measurement setup. Surface characterization using quartz crystal microbalance (QCM) and high-resolution atomic force microscope (AFM) is used to benchmark the cantilever sensor response. In addition, the QCM and AFM studies reveal that the surface density of bound streptavidin on biotin modified surfaces was low, thereby implying that effects other than steric hindrance are responsible for defining cantilever response. (c) 2006 Elsevier B.V. All rights reserved.

Transfer of dynamic learning across postures.

When learning a difficult motor task, we often decompose the task so that the control of individual body segments is practiced in isolation. But on re-composition, the combined movements can result in novel and possibly complex internal forces between the body segments that were not experienced (or did not need to be compensated for) during isolated practice. Here we investigate whether dynamics learned in isolation by one part of the body can be used by other parts of the body to immediately predict and compensate for novel forces between body segments. Subjects reached to targets while holding the handle of a robotic, force-generating manipulandum. One group of subjects was initially exposed to the novel robot dynamics while seated and was then tested in a standing position. A second group was tested in the reverse order: standing then sitting. Both groups adapted their arm dynamics to the novel environment, and this movement learning transferred between seated and standing postures and vice versa. Both groups also generated anticipatory postural adjustments when standing and exposed to the force field for several trials. In the group that had learned the dynamics while seated, the appropriate postural adjustments were observed on the very first reach on standing. These results suggest that the CNS can immediately anticipate the effect of learned movement dynamics on a novel whole-body posture. The results support the existence of separate mappings for posture and movement, which encode similar dynamics but can be adapted independently.

Flexible representations of dynamics are used in object manipulation.

To manipulate an object skillfully, the brain must learn its dynamics, specifying the mapping between applied force and motion. A fundamental issue in sensorimotor control is whether such dynamics are represented in an extrinsic frame of reference tied to the object or an intrinsic frame of reference linked to the arm. Although previous studies have suggested that objects are represented in arm-centered coordinates [1-6], all of these studies have used objects with unusual and complex dynamics. Thus, it is not known how objects with natural dynamics are represented. Here we show that objects with simple (or familiar) dynamics and those with complex (or unfamiliar) dynamics are represented in object- and arm-centered coordinates, respectively. We also show that objects with simple dynamics are represented with an intermediate coordinate frame when vision of the object is removed. These results indicate that object dynamics can be flexibly represented in different coordinate frames by the brain. We suggest that with experience, the representation of the dynamics of a manipulated object may shift from a coordinate frame tied to the arm toward one that is linked to the object. The additional complexity required to represent dynamics in object-centered coordinates would be economical for familiar objects because such a representation allows object use regardless of the orientation of the object in hand.

Correct biological timing in Arabidopsis requires multiple light-signaling pathways.

Circadian oscillators provide rhythmic temporal cues for a range of biological processes in plants and animals, enabling anticipation of the day/night cycle and enhancing fitness-associated traits. We have used engineering models to understand the control principles of a plant's response to seasonal variation. We show that the seasonal changes in the timing of circadian outputs require light regulation via feed-forward loops, combining rapid light-signaling pathways with entrained circadian oscillators. Linear time-invariant models of circadian rhythms were computed for 3,503 circadian-regulated genes and for the concentration of cytosolic-free calcium to quantify the magnitude and timing of regulation by circadian oscillators and light-signaling pathways. Bioinformatic and experimental analysis show that rapid light-induced regulation of circadian outputs is associated with seasonal rephasing of the output rhythm. We identify that external coincidence is required for rephasing of multiple output rhythms, and is therefore important in general phase control in addition to specific photoperiod-dependent processes such as flowering and hypocotyl elongation. Our findings uncover a fundamental design principle of circadian regulation, and identify the importance of rapid light-signaling pathways in temporal control.

Anharmonic surface interactions for biomolecular screening and characterization.

The acoustic response of conventional mechanical oscillators, such as a piezoelectric crystal, is predominantly harmonic at modest amplitudes. However, here, we observe from the electrical response that significant motional anharmonicity is introduced in the presence of attached analyte. Experiments were conducted with streptavidin-coated polystyrene microbeads of various sizes attached to a quartz crystal resonator via specific and nonspecific molecular tethers in liquid. Quantitative analysis reveals that the deviation of odd Fourier harmonics of the response caused by introduction of microbeads as a function of oscillation amplitude presents a unique signature of the molecular tether. Hence, the described anharmonic detection technique (ADT) based on this function allows screening of biomolecules and provides an additional level of selectivity in receptor-based detection that is often associated with nonspecific interactions. We also propose methods to extract mechanical force-extension characteristics of the molecular tether and activation energy using this technique.

Integration of holographic sensors into microfluidics for the real-time pH sensing of L. casei metabolism

We describe developments in the integration of analyte specific holographic sensors into PDMS-based microfluidic devices for the purpose of continuous, low-impact monitoring of extra-cellular change in micro-bioreactors. Holographic sensors respond to analyte concentration via volume change, which makes their reduction in size and integration into spatially confined fluidics difficult. Through design and process modification many of these constraints have been addressed, and a microfluidics-based device capable of real-time monitoring of the pH change caused by Lactobacillus casei fermentation is presented as a general proof-of-concept for a wide array of possible devices.

Microbead dynamics on quartz crystal microbalance at elevated amplitudes

The application of the Quartz Crystal Microbalance (QCM) for biochemical sensing is well known. However, utilizing the nonlinear response of the QCM at elevated amplitudes has received sporadic attention. This study presents results for QCM-analyte interaction that provide insight into the nonlinear dynamics of the QCM with attached analyte. In particular, interactions of the QCM with polystyrene microbeads physisorbed via self-assembled monolayer (SAM) were studied through experiments and modelling. It was found that the response of the QCM coupled to these surface adsorbents is anharmonic even at low oscillation amplitudes and that the nonlinear signals from such interactions are much higher than those for bare quartz. Therefore, these signals can potentially be used as sensitive signatures of adsorbents and their kinetics on the surface. ©2009 IEEE.

GSPS photonic analogue to digital conversion system using broadband continuous wave source

An 80 GSPS photonic ADC system is demonstrated, using broadband MLL and dispersive fibre to form a continuous waveform with time-wavelength mapping, and AWG to channelise. Tests are carried out for RF signals up to 10GHz. © 2005 Optical Society of America.

Effects of mechanical and electrical coupling on the parametric sensitivity of mode localized sensors

We compare and contrast the effects of two distinctly different mechanisms of coupling (mechanical and electrical) on the parametric sensitivity of micromechanical sensors utilizing mode localization for sensor applications. For the first time, the strong correlation between mode localization and the phenomenon of 'eigenvalue loci-veering' is exploited for accurate quantification of the strength of internal coupling in mode localized sensors. The effects of capacitive coupling-spring tuning on the parametric sensitivity of electrically coupled resonators utilizing this sensing paradigm is also investigated and a mass sensor with sensitivity tunable by over 400% is realized. ©2009 IEEE.

The circadian oscillator gene GIGANTEA mediates a long-term response of the Arabidopsis thaliana circadian clock to sucrose.

Circadian clocks are 24-h timing devices that phase cellular responses; coordinate growth, physiology, and metabolism; and anticipate the day-night cycle. Here we report sensitivity of the Arabidopsis thaliana circadian oscillator to sucrose, providing evidence that plant metabolism can regulate circadian function. We found that the Arabidopsis circadian system is particularly sensitive to sucrose in the dark. These data suggest that there is a feedback between the molecular components that comprise the circadian oscillator and plant metabolism, with the circadian clock both regulating and being regulated by metabolism. We used also simulations within a three-loop mathematical model of the Arabidopsis circadian oscillator to identify components of the circadian clock sensitive to sucrose. The mathematical studies identified GIGANTEA (GI) as being associated with sucrose sensing. Experimental validation of this prediction demonstrated that GI is required for the full response of the circadian clock to sucrose. We demonstrate that GI acts as part of the sucrose-signaling network and propose this role permits metabolic input into circadian timing in Arabidopsis.