Biomaterial immobilization on polyurethane films by XeCl excimer laser processing

The surface chemical modification of polyurethane (PU) films was performed by an UV laser-induced chemical reaction in a polysaccharide solution. This process may be applicable as hydrophilic packaging of implantable medical devices and in vivo sensors. When a PU film in contact with an aqueous alginic acid (AAC) solution was irradiated with a XeCl laser, the PU film turned hydrophilic. Contact angles of water on the film were reduced from 110° to 60°. Since light absorption of the AAC solution at 308 nm was negligibly small, reactive sites were generated solely on the PU surface. There, AAC could be immobilized by chemical bonds thus allowing for a nanometer-scaled grafting of this biomolecule. The mechanism was investigated by surface analyses with Fourier-transform infrared spectroscopy (FT-IR), dye staining, ultraviolet-visible (UV-VIS) spectroscopy, and scanning electron microscopy (SEM) techniques. A one-photon photochemical process could beidentified. Received: 30 June 2000 / Accepted: 4 July 2000 / Published online: 13 September 2000     

Amplitude noise suppression in high-repetition-rate pulse train generation from a frequency-modulated Er-Yb laser

A detailed experimental analysis of the amplitude noise in high-repetition-rate picosecond pulse trains generated by spectral filtering of a frequency-modulated Er-Yb:glass laser is reported. Two distinct sources of noise are identified, and stabilization techniques for noise suppression are proposed and experimentally demonstrated. Intensity noise suppression of ∼20 dB, corresponding to less than 2% residual amplitude fluctuations of the pulse train, has been achieved at repetition rates of 2.5 GHz and 5 GHz with pulse durations of ∼50 ps. Received: 28 July 1999 / Revised version: 13 September 1999 / Published online: 20 October 1999     

A model for the self-organization of microtubules driven by molecular motors

We propose a two-dimensional model for the organization of stabilized microtubules driven by molecular motors in an unconfined geometry. In this model two kinds of dynamics are competing. The first one is purely diffusive, with an interaction between the rotational degrees of freedom, while the second one is a local drive, dependent on microtubule polarity. As a result, there is a configuration dependent driving field. Applying a molecular field approximation, we are able to derive continuum equations. A study on the solutions of these equations shows non-equilibrium inhomogeneous steady states in various regions of the parameter space. The presence and stability of such self-organized states are investigated in terms of entropy production. Numerical simulations confirm our analytic results. Received 4 August 1999 and Received in final form 24 November 1999     

Anisotropic elastic properties of microtubules

The present study aims at providing insight into the acceleration mechanism of a bubble chain rising in shear-thinning viscoelastic fluids. The experimental investigation by the Particle Image Velocimetry (PIV), birefringence visualisation and rheological simulation shows that two aspects are central to bubble interactions in such media: the stress creation by the passage of bubbles, and their relaxation due to the fluids memory forming an evanescent corridor of reduced viscosity. Interactions between bubbles were taken into account mainly through a linear superposition of the stress evolution behind each bubble. An analytical approach together with the rheological consideration was developed to compute the rise velocity of a bubble chain in function of the injection period and bubble volume. The model predictions compare satisfactorily with the experimental investigation.     

Ratchet patterns sort molecular shuttles

Molecular shuttles based on microtubules propelled by motor proteins can be guided on surfaces by adsorbing motors in chemical patterns or by using open guiding channels. While chemical patterns can guide microtubules based on a Brownian ratchet mechanism, the rigidity of the microtubules limits guiding to features with dimensions on the order of their persistence length (5 mm). To achieve guiding on micron-scale dimensions, physical barriers are required which can exploit the forces exerted by multiple motors to bend tubules into tight radii of curvature. Microtubule guiding is illustrated for the case of a special ratchet pattern that is capable of sorting microtubules on the basis of the direction of their motion. Received: 3 December 2001 / Accepted: 11 February 2002 / Published online: 22 April 2002     

Strategies for the separation of polyelectrolytes based on non-linear dynamics and entropic ratchets in a simple microfluidic device

We perform Monte Carlo simulations of an existing electrophoretic microchannel device used for the size separation of large DNA fragments. This device is normally operated with a constant (dc) driving field. In contrast, we consider the case of a varying (ac) driving field, in the zero-frequency limit. We find that a time-asymmetric pulse can yield interesting migration regimes, in particular bidirectional transport for different molecular sizes. We also study a spatially asymmetric version of the device and show that it can rectify unbiased but non-equilibrium molecular motion, in agreement with previous predictions for entropic ratchets. Finally, at finite frequency we uncover a resonance for the molecular velocity in the channel which could lead to improved performance. Received: 16 November 2001 / Accepted: 11 February 2002 / Published online: 22 April 2002     

Optical tweezers for confocal microscopy

In confocal laser scanning microscopes (CLSMs), lasers can be used for image formation as well as tools for the manipulation of microscopic objects. In the latter case, in addition to the imaging lasers, the light of an extra laser has to be focused into the object plane of the CLSM, for example as optical tweezers. Imaging as well as trapping by optical tweezers can be done using the same objective lens. In this case, z-sectioning for 3D imaging shifts the optical tweezers with the focal plane of the objective along the optical axis, so that a trapped object remains positioned in the focal plane. Consequently, 3D imaging of trapped objects is impossible without further measures. We present an experimental set-up keeping the axial trapping position of the optical tweezers at its intended position whilst the focal plane can be axially shifted over a distance of about 15 μm. It is based on fast-moving correctional optics synchronized with the objective movement. First examples of application are the 3D imaging of chloroplasts of Elodea densa (Canadian waterweed) in a vigorous cytoplasmic streaming and the displacement of zymogen granules in pancreatic cancer cells (AR42 J). Received: 24 March 2000 / Revised version: 23 June 2000 / Published online: 11 October 2000     

AFM, a tool for single-molecule experiments

Received: 27 March 1998     

Electrostatic theory of viral self-assembly

Viruses self-assemble from identical capsid proteins and their genome consisting, for example, of a long single stranded (ss) RNA. For a big class of T=3 viruses, capsid proteins have long positive N-terminal tails. We explore the role played by the Coulomb interaction between the brush of positive N-terminal tails rooted at the inner surface of the capsid and the negative ss RNA molecule. We show that viruses are most stable when the total contour length of ss RNA is close to the total length of the tails. For such a structure the absolute value of the total RNA charge is approximately twice as large as the charge of the capsid. This conclusion agrees with structural data.     

Photobleaching and stabilization of. fluorophores used for single-molecule analysis. with one- and two-photon excitation

Fluorescence spectroscopic measurements at the single-molecule level usually require large absorption cross sections and fluorescence quantum yields for the dyes under study. In addition to these parameters, the collectable number of fluorescence photons and, thus, the signal-to-noise ratio of the measurement, is influenced by processes like triplet-state population and photobleaching, shifting the saturation threshold of the dye to lower excitation intensities. Confocal fluorescence correlation spectroscopy (FCS) is a versatile method to precisely determine photon emission rates of single molecules but also gives access to rate constants of dynamic bleaching and intersystem crossing. In recent FCS studies in solution and living cells, two-photon excitation with its inherent spatial sectioning has proven to be a very valuable alternative to minimize background and cumulative signal loss. However, there is evidence that in many dye systems, the photobleaching rates with two-photon excitation are significantly enhanced with respect to one-photon excitation at comparable photon-emission yields. The reasons have so far remained mainly speculative. In the present study, potential photobleaching pathways are investigated by adding chemical stabilizers and by working at different oxygen concentrations. The results suggest that the population of triplet states does not appear to be responsible for the limited emission rate with two-photon excitation. Rather, photobleaching pathways via the formation of radicals seem to be plausible causes for the signal limitation. Favorable conditions are discussed to maximize the overall photon-collection yield in two-photon experiments. Received: 4 July 2001 / Published online: 23 November 2001     

Ultra-broadband continuum generation by hollow-fiber cascading

A novel spectral broadening technique, based on hollow-fiber cascading, is reported, which allows the generation of a supercontinuum extending to a bandwidth exceeding 510 THz with excellent spatial beam quality. High-peak-power, sub-7-fs light pulses tunable from the visible to the near infrared have been generated by compression of portions of the supercontinuum, employing different sets of chirped mirrors. Received: 11 July 2002 / Revised version: 3 September 2002 / Published online: 15 November 2002 RID="*" ID="*"Corresponding author. Fax: +39-02/2399-6126, E-mail: mauro.nisoli@fisi.polimi.it     

Analytical functions for the optimization of second-harmonic generation and parametric generation by focused Gaussian beams

An analytical function involving four parameters is proposed to express the second-harmonic generation efficiency as well as the parametric generation gain coefficient in the Boyd–Kleinman theory. The analytical function clearly reveals the dependence of conversion efficiency on the focusing parameter and the walk-off parameter. Moreover, the optimum focusing parameter and its corresponding maximum efficiency are explicitly given in the analytical function, leading to a straightforward evaluation of a given crystal performance. Received: 20 January 2003 / Published online: 22 May 2003 RID="*" ID="*"Corresponding author. Fax: +886-35/7291-34, E-mail: yfchen@cc.nctu.edu.tw     

Simulations of flexible membranes using a coarse-grained particle-based model with spontaneous curvature variables

We propose a simple mathematical model for lipid bilayer membranes of flexible fluid or solid state. The model consists of interacting coarse-grained particles with two extra variables. One indicates the spontaneous curvature at the particle position, and the other indicates the vector representing the direction normal to the membrane. When the spontaneous curvature variable is allowed to fluctuate significantly, the fluctuation causes a softening of the membrane and growth of large undulations as the amplitude of the fluctuation is increased. By changing the amplitude of the fluctuation in simulations, the bending rigidity of the membrane can be easily controlled. Because the proposed model includes anisotropic interactions between the particles, multilayered vesicles can be obtained through a reversible transition by weakening the strength of the anisotropic interactions.     

Slow nucleic acid unzipping kinetics from sequence-defined barriers

Recent experiments on unzipping of RNA helix-loop structures by force have shown that ≈40-base molecules can undergo kinetic transitions between two well-defined “open” and “closed” states, on a timescale ≈1 sec [Liphardt et al., Science 297, 733-737 (2001)]. Using a simple dynamical model, we show that these phenomena result from the slow kinetics of crossing large free energy barriers which separate the open and closed conformations. The dependence of barriers on sequence along the helix, and on the size of the loop(s) is analyzed. Some DNA and RNA sequences that could show dynamics on different time scales, or three(or more)-state unzipping, are proposed. Our dynamical model is also applied to the unzipping of long (kilo-basepair) DNA molecules at constant force. Received 29 July 2002 / Received in final form 5 February 2003 Published online: 16 April 2003 RID="a" ID="a"e-mail: cocco@ldfc.u-strasbg.fr RID="b" ID="b"e-mail: jmarko@uic.edu RID="c" ID="c"e-mail: monasson@lpt.ens.fr     

Membrane transistor with giant lipid vesicle touching a silicon chip

Lipid bilayers on silicon may become the matrix of future bioelectronic devices if the junction is sufficiently insulating. We touched the open gate of a field-effect transistor with a preformed giant lipid vesicle and bound the membrane by means of polyelectrolyte interaction. The sheet resistance along the junction was 100 GΩ and the membrane resistance was above 100 GΩ at a contact area of 1000 μm². The bilayer was fluid and smoothly followed the surface profile of the chip. The compound lipid–silicon structure is suitable to couple semiconductor and electroactive proteins. Received: 12 August 1999 / Accepted: 16 August 1999 / Published online: 6 October 1999     

Force spectroscopy on single xanthan molecules

Received: 3 July 1998 / Published online: 10 February 1999     

Far-field distribution of third-harmonic generation by two crossed beams

The far-field intensity distribution of the third harmonic generated in a thin silica plate by two laser beams crossed at a small angle comprises four peaks approximately symmetric with respect to the bisector. The measurement of the intensity ratio of the inner to the outer peaks yielded values scattered from 10 to 20, surprisingly larger than the combinatorial value 9. The phase-matching factor, as determined by the full three-dimensional calculations required by the absence of axial symmetry, accounts for the observed ratio and its fluctuations. Received: 14 November 2002 / Published online: 22 May 2003 RID="*" ID="*"Corresponding author. Fax: +7-95/1941-994, E-mail: fomichev@imp.kiae.ru     

Tracking of fluorescent molecules diffusing within membranes

A new method is proposed for tracking fluorescing single molecules diffusing within a two-dimensional membrane. It is based on a confocal microscopy setup with a constantly rotating laser focus, which follows the position of the molecule. The optimization and efficiency of the method are theoretically studied for a broad range of experimentally realistic conditions. The proposed method allows for a long-time observation of diffusing molecules while allowing the application of fast spectroscopic techniques such as fluorescence decay time determination or fluorescence anisotropy measurements. Received: 14 January 2000 / Published online: 13 September 2000