Optimizing sampling for surface localization in 3D-scanning microscopy.

3D-scanning fluorescence imaging of living tissue is in demand for less phototoxic acquisition process. For the imaging of biological surfaces, adaptive and sparse scanning schemes have been proven to efficiently reduce the light dose by concentrating acquisitions around the surface. In this paper, we focus on optimizing the scanning scheme at a constant photon budget, when the problem is to estimate the position of a biological surface whose intensity profile is modeled as a Gaussian shape. We propose an approach based on the Cramér-Rao bound to optimize the positions and number of scanning points, assuming signal-dependant Gaussian noise. We show that, in the case of regular sampling, the optimization problem can be reduced to a few parameters, allowing us to define quasi-optimal acquisition strategies, first when no prior knowledge of the surface location is available and then when the user has a prior on this location.

Orchestrating size and shape during morphogenesis.

Living organisms exhibit tremendous diversity, evident in the large repertoire of forms and considerable size range. Scientists have discovered that conserved mechanisms control the development of all organisms. Drosophila has proved to be a particularly powerful model system with which to identify the signalling pathways that organize tissue patterns. More recently, much has been learned about the control of tissue growth, tissue shape and their coordination at the cellular and tissue levels. New models integrate how specific signals and mechanical forces shape tissues and may also control their size.

Adaptive shift in the domain of negative stiffness during spontaneous oscillation by hair bundles from the internal ear.

When a hair cell of the bullfrog's sacculus is maintained in vitro under native ionic conditions, its mechanosensitive hair bundle may oscillate spontaneously. This movement has been hypothesized to result from the interaction of the bundle's negative stiffness, which creates a region of mechanical instability, with a myosin-based adaptation mechanism that continually repositions the bundle there. To test this proposition, we used a flexible stimulus fiber in an analog feedback loop to measure the displacement-force relation of an oscillating hair bundle. A digital signal processor was used to monitor spontaneous oscillations in real time and trigger measurements at particular phases of the movement cycle. By comparing the displacement-force curves obtained at the two extremes of a hair bundle's motion, we demonstrated a shift in the negative-stiffness region whose direction, orientation, magnitude, and kinetics agreed with the predictions of the gating-spring theory. The results are in accordance with the idea that adaptation underlies spontaneous hair-bundle oscillation, and therefore powers the active process that also amplifies and tunes the hair cell's mechanical responsiveness.

Tracer studies on f-actin fluctuations.

We present a study on the fluctuations of semiflexible actin filaments using fluorescence videomicroscopy, focusing on the end-to-end fluctuations of single filaments. In order to specifically measure the position of the polymer's ends, we developed a novel noninvasive method that consists of annealing short end tags to the filaments. This allows us to probe polymer fluctuations to a very high accuracy. We compared the distribution of the end-to-end distance with recent theoretical results, and found excellent agreement. We also studied the dynamics of the mean-square end-to-end distance deltaR2(t) and orientation of the ends, deltaTheta(2)(t), finding power laws t(3/4) and t(1/4), respectively. Scaling behavior for deltaR2(t) is observed over several decades in relaxation time in agreement with theoretical results.

Motor-driven dynamics in actin-myosin networks.

The effect of myosin motor protein activity on the filamentous actin (F-actin) rheological response is studied using diffusing wave spectroscopy. Under conditions of saturating motor activity, we find an enhancement of longitudinal filament fluctuations corresponding to a scaling of the viscoelastic shear modulus G(d)(omega) approximately omega(7/8). As the adenosine tri-phosphate reservoir sustaining motor activity is depleted, we find an abrupt transient to a passive, "rigor state" and a return to dissipation dominated by transverse filament modes. Single-filament measurements of the apparent persistence length support the notion that motor activity leads to an increase in the effective temperature for tangential motion.