Searcharxiv⌕ Search

arXiv subjects

Felix Ritort

Publications and source records attributed to Felix Ritort.

At least 19 recordsLinked to original sources
arXiv2025

SmartTrap: Automated Precision Experiments with Optical Tweezers

There is a trend in research towards more automation using smart systems powered by artificial intelligence. While experiments are often challenging to automate, they can greatly benefit from automation by reducing labor and increasing reproducibility. For example, optical tweezers are widely employed in single-molecule biophysics, cell biomechanics, and soft matter physics, but they still require a human operator, resulting in low throughput and limited repeatability. Here, we present a smart optical tweezers platform, which we name SmartTrap, capable of performing complex experiments completely autonomously. SmartTrap integrates real-time 3D particle tracking using deep learning, custom electronics for precise feedback control, and a microfluidic setup for particle handling. We demonstrate the ability of SmartTrap to operate continuously, acquiring high-precision data over extended periods of time, through a series of experiments. By bridging the gap between manual experimentation and autonomous operation, SmartTrap establishes a robust and open source framework for the next generation of optical tweezers research, capable of performing large-scale studies in single-molecule biophysics, cell mechanics, and colloidal science with reduced experimental overhead and operator bias.

physics.bio-ph↗
arXiv2024

Stacking correlation length in single-stranded DNA

Base stacking is crucial in nucleic acid stabilization, from DNA duplex hybridization to single-stranded DNA (ssDNA) protein binding. While stacking energies are tiny in ssDNA, they are inextricably mixed with hydrogen bonding in DNA base pairing, making their measurement challenging. We conduct unzipping experiments with optical tweezers of short poly-purine (dA and alternating dG and dA) sequences of 20-40 bases. We introduce a helix-coil model of the stacking-unstacking transition that includes finite length effects and reproduces the force-extension curves. Fitting the model to the experimental data, we derive the stacking energy per base, finding the salt-independent value $\Delta G_0$ = 0.14(3) kcal/mol for poly-dA and $\Delta G_0$ = 0.07(3) kcal/mol for poly-dGdA. Stacking in these polymeric sequences is predominantly cooperative with a correlation length of $\sim4$ bases at zero force. The correlation length reaches a maximum of $\sim10$ and 5 bases at the stacking-unstacking transition force of $\sim10$ and 20 pN for poly-dA and poly-dGdA, respectively. The salt dependencies of the cooperativity parameter in ssDNA and the energy of DNA hybridization are in agreement, suggesting that double-helix stability is primarily due to stacking. Analysis of poly-rA and poly-rC RNA sequences shows a larger stacking stability but a lower stacking correlation length of $\sim2$ bases.

cond-mat.soft↗
arXiv2024

DNA Calorimetric Force Spectroscopy at Single Base Pair Resolution

DNA hybridization is a fundamental reaction with wide-ranging applications in biotechnology. The nearest-neighbor (NN) model provides the most reliable description of the energetics of duplex formation. Most DNA thermodynamics studies have been done in melting experiments in bulk, of limited resolution due to ensemble averaging. In contrast, single-molecule methods have reached the maturity to derive DNA thermodynamics with unprecedented accuracy. We combine single-DNA mechanical unzipping experiments using a temperature jump optical trap with machine learning methods and derive the temperature-dependent DNA energy parameters of the NN model. In particular, we measure the previously unknown ten heat-capacity change parameters $\Delta C_p$, relevant for thermodynamical predictions throughout the DNA stability range. Calorimetric force spectroscopy establishes a groundbreaking methodology to accurately study nucleic acids, from chemically modified DNA to RNA and DNA/RNA hybrid structures.

q-bio.BM↗
arXiv2024

Universal Cold RNA Phase Transitions

RNA's diversity of structures and functions impacts all life forms since primordia. We use calorimetric force spectroscopy to investigate RNA folding landscapes in previously unexplored low-temperature conditions. We find that Watson-Crick RNA hairpins, the most basic secondary structure elements, undergo a glass-like transition below $\mathbf{T_G\sim 20 ^{\circ}}$C where the heat capacity abruptly changes and the RNA folds into a diversity of misfolded structures. We hypothesize that an altered RNA biochemistry, determined by sequence-independent ribose-water interactions, outweighs sequence-dependent base pairing. The ubiquitous ribose-water interactions lead to universal RNA phase transitions below $\mathbf{T_G}$, such as maximum stability at $\mathbf{T_S\sim 5 ^{\circ}}$C where water density is maximum, and cold denaturation at $\mathbf{T_C\sim-50^{\circ}}$C. RNA cold biochemistry may have a profound impact on RNA function and evolution.

q-bio.BM↗
arXiv2024

Variance sum rule: proofs and solvable models

We derive, in more general conditions, a recently introduced variance sum rule (VSR) [I. Di Terlizzi et al., 2024 Science 383 971] involving variances of displacement and force impulse for overdamped Langevin systems in a nonequilibrium steady state (NESS). This formula allows visualising the effect of nonequilibrium as a deviation of the sum of variances from normal diffusion $2Dt$, with $D$ the diffusion constant and $t$ the time. From the VSR, we also derive formulas for the entropy production rate $\sigma$ that, differently from previous results, involve second-order time derivatives of position correlation functions. This novel feature gives a criterion for discriminating strong nonequilibrium regimes without measuring forces. We then apply and discuss our results to three analytically solved models: a stochastic switching trap, a Brownian vortex, and a Brownian gyrator. Finally, we compare the advantages and limitations of known and novel formulas for $\sigma$ in an overdamped NESS.

cond-mat.stat-mech↗
arXiv2023

Detecting molecular folding from noise measurements

Detecting conformational transitions in molecular systems is key to understanding biological processes. Here, we investigate the force variance in single-molecule pulling experiments as an indicator of molecular folding transitions. We consider cases where Brownian force fluctuations are large, masking the force rips and jumps characteristic of conformational transitions. We compare unfolding and folding data for DNA hairpin systems of loop sizes 4,8, and 20 and the 110 amino acids protein barnase, finding conditions that facilitate the detection of folding events at low forces where the signal-to-noise ratio is low. In particular, we discuss the role of temperature as a useful parameter to improve the detection of folding transitions in entropically driven processes where folding forces are temperature-independent. The force variance approach might be extended to detect the elusive intermediate states in RNA and protein folding.

cond-mat.stat-mech↗
arXiv2023

N-States Continuous Maxwell Demon

Maxwell's demon is a famous thought experiment and a paradigm of the thermodynamics of information. It is related to Szilard's engine, a two-state information-to-work conversion device in which the demon performs single measurements and extracts work depending on the state measurement outcome. A variant of these models, the Continuous Maxwell Demon (CMD), was recently introduced by Ribezzi-Crivellari and Ritort where work was extracted every time $\tau$ in a two state model. The CMD was able to extract unbounded amounts of work at the cost of an unbounded amount of information storage. In this work, we built a generalization of the CMD to the N-states case. We obtained generalized analytical expressions for the average work extracted and the information content. We show that the second law inequality for information-to-work conversion is fulfilled. We illustrate the results for N-states with uniform transition rates and for the N=3 case.

cond-mat.stat-mech↗
arXiv2023

Unraveling RNA by Mechanical Unzipping

We review the basic concepts and tools for mechanically unzipping RNA hairpins using force spectroscopy. By pulling apart the ends of an RNA molecule using optical tweezers, it is possible to measure the folding free energy at varying experimental conditions. Energy measurements permit us to characterize the thermodynamics of RNA hybridization (base pairing and stacking), the dynamics of the formation of native and kinetic (intermediates and misfolded) molecular states, and interactions with metallic ions. This paper introduces basic concepts and reviews recent developments related to RNA force thermodynamics, native and barrier RNA energy landscapes, and RNA folding dynamics. We emphasize the implications of mechanical unzipping experiments to understand non-coding RNAs and RNAs in extreme environments.

cond-mat.soft↗
arXiv2022

Towards a robust criterion of anomalous diffusion

Anomalous-diffusion, the departure of the spreading dynamics of diffusing particles from the traditional law of Brownian-motion, is a signature feature of a large number of complex soft-matter and biological systems. Anomalous-diffusion emerges due to a variety of physical mechanisms, e.g., trapping interactions or the viscoelasticity of the environment. However, sometimes systems dynamics are erroneously claimed to be anomalous, despite the fact that the true motion is Brownian -- or vice versa. This ambiguity in establishing whether the dynamics as normal or anomalous can have far-reaching consequences, e.g., in predictions for reaction- or relaxation-laws. Demonstrating that a system exhibits normal- or anomalous-diffusion is highly desirable for a vast host of applications. Here, we present a criterion for anomalous-diffusion based on the method of power-spectral analysis of single trajectories. The robustness of this criterion is studied for trajectories of fractional-Brownian-motion, a ubiquitous stochastic process for the description of anomalous-diffusion, in the presence of two types of measurement errors. In particular, we find that our criterion is very robust for subdiffusion. Various tests on surrogate data in absence or presence of additional positional noise demonstrate the efficacy of this method in practical contexts. Finally, we provide a proof-of-concept based on diverse experiments exhibiting both normal and anomalous-diffusion.

cond-mat.stat-mech↗
arXiv2022

Experimental test of Sinai's model in DNA unzipping

The experimental measurement of correlation functions and critical exponents in disordered systems is key to testing renormalization group (RG) predictions. We mechanically unzip single DNA hairpins with optical tweezers, an experimental realization of the diffusive motion of a particle in a one-dimensional random force field, known as the Sinai model. We measure the unzipping forces $F_w$ as a function of the trap position $w$ in equilibrium and calculate the force-force correlator $\Delta_m(w)$, its amplitude, and correlation length, finding agreement with theoretical predictions. We study the universal scaling properties since the effective trap stiffness $m^2$ decreases upon unzipping. Fluctuations of the position of the base pair at the unzipping junction $u$ scales as $u \sim m^{-\zeta}$, with a roughness exponent $ \zeta=1.34\pm0.06$, in agreement with the analytical prediction $\zeta = \frac{4}{3}$. Our study provides a single-molecule test of the functional RG approach for disordered elastic systems in equilibrium.

cond-mat.dis-nn↗
arXiv2022

Roadmap for Optical Tweezers

Optical tweezers are tools made of light that enable contactless pushing, trapping, and manipulation of objects ranging from atoms to space light sails. Since the pioneering work by Arthur Ashkin in the 1970s, optical tweezers have evolved into sophisticated instruments and have been employed in a broad range of applications in life sciences, physics, and engineering. These include accurate force and torque measurement at the femtonewton level, microrheology of complex fluids, single micro- and nanoparticle spectroscopy, single-cell analysis, and statistical-physics experiments. This roadmap provides insights into current investigations involving optical forces and optical tweezers from their theoretical foundations to designs and setups. It also offers perspectives for applications to a wide range of research fields, from biophysics to space exploration.

physics.optics↗
arXiv2022

Force-Dependent Folding Kinetics of Single Molecules with Multiple Intermediates and Pathways

Most single-molecule studies derive the kinetic rates of native, intermediate, and unfolded states from equilibrium hopping experiments. Here, we apply Kramers kinetic diffusive model to derive the force-dependent kinetic rates of intermediate states from non-equilibrium pulling experiments. From the kinetic rates, we also extract the force-dependent kinetic barriers and the equilibrium folding energies. We apply our method to DNA hairpins with multiple folding pathways and intermediates. The experimental results agree with theoretical predictions. Furthermore, the proposed non-equilibrium single-molecule approach permits us to characterize kinetic and thermodynamic properties of native, unfolded, and intermediate states that cannot be derived from equilibrium hopping experiments.

cond-mat.soft↗
arXiv2022

Molten globule-like transition state of protein barnase measured with calorimetric force spectroscopy

Understanding how proteins fold into their native structure is a fundamental problem in biophysics, crucial for protein design. It has been hypothesized that the formation of a molten globule intermediate precedes folding to the native conformation of globular proteins; however, its thermodynamic properties are poorly known. We perform single-molecule pulling experiments of protein barnase in the range of 7$^\circ$C to 37$^\circ$C using a temperature-jump optical trap. We derive the folding free energy, entropy and enthalpy, and heat capacity change ($\Delta C_p = 1050\pm50$ cal/mol$\cdot$K) at low ionic strength conditions. From the measured unfolding and folding kinetic rates, we also determine the thermodynamic properties of the transition state, finding a significant change in $\Delta C_p$ ($\sim$ 90$\%$) between the unfolded and the transition states. In contrast, the major change in enthalpy ($\sim$ 80$\%$) occurs between the transition and native states. These results highlight a transition state of high energy and low configurational entropy structurally similar to the native state, in agreement with the molten globule hypothesis.

cond-mat.soft↗
arXiv2022

Stem-loop formation drives RNA folding in mechanical unzipping experiments

Accurate knowledge of RNA hybridization is essential for understanding RNA structure and function. Here we mechanically unzip and rezip a 2-kbp RNA hairpin and derive the 10 nearest-neighbor base pair (NNBP) RNA free energies in sodium and magnesium with 0.1 kcal/mol precision using optical tweezers. Notably, force-distance curves (FDCs) exhibit strong irreversible effects with hysteresis and several intermediates, precluding the extraction of the NNBP energies with currently available methods. The combination of a suitable RNA synthesis with a tailored pulling protocol allowed us to obtain the fully reversible FDCs necessary to derive the NNBP energies. We demonstrate the equivalence of sodium and magnesium free-energy salt corrections at the level of individual NNBP. To characterize the irreversibility of the unzipping-rezipping process, we introduce a barrier energy landscape of the stem-loop structures forming along the complementary strands, which compete against the formation of the native hairpin. This landscape correlates with the hysteresis observed along the FDCs. RNA sequence analysis shows that base stacking and base pairing stabilize the stem-loops that kinetically trap the long-lived intermediates observed in the FDC. Stem-loops formation appears as a general mechanism to explain a wide range of behaviors observed in RNA folding.

q-bio.BM↗
arXiv2021

Dissipation reduction and information-to-measurement conversion in DNA pulling experiments with feedback protocols

Information-to-energy conversion with feedback measurement stands as one of the most intriguing aspects of the thermodynamics of information in the nanoscale. To date, experiments have focused on feedback protocols for work extraction. Here we address the novel case of dissipation reduction in non-equilibrium systems with feedback. We perform pulling experiments on DNA hairpins with optical tweezers, with a general feedback protocol based on multiple measurements that includes either discrete-time or continuous-time feedback. While feedback can reduce dissipation, it remains unanswered whether it also improves free energy determination (information-to-measurement conversion). We define thermodynamic information {\Upsilon} as the natural logarithm of the feedback efficacy, a quantitative measure of the efficiency of information-to-energy and information-to-measurement conversion in feedback protocols. We find that discrete and continuous-time feedback reduces dissipation by roughly kBT{\Upsilon} without improvement in free energy determination. Remarkably, a feedback strategy (defined as a correlated sequence of feedback protocols) further reduces dissipation, enhancing information-to-measurement efficiency. Our study underlines the role of temporal correlations to develop feedback strategies for efficient information-to-energy conversion in small systems.

cond-mat.stat-mech↗
arXiv2021

Information-to-work conversion in single molecule experiments: from discrete to continuous feedback

We theoretically investigate the extractable work in single molecule unfolding-folding experiments with applied feedback. Using a simple two-state model, we obtain a description of the full work distribution, from discrete to continuous feedback. The effect of the feedback is captured by a detailed fluctuation theorem, accounting for the information aquired. We find analytical expressions for the average work extraction as well as an experimentally measurable bound thereof, which becomes tight in the continuous feedback limit. We further determine the parameters for maximal power, or rate of work extraction. While our two-state model only depends on a single, effective transition rate, we find quantitative agreement with Monte Carlo simulations of DNA hairpin unfolding-folding dynamics.

cond-mat.stat-mech↗
arXiv2021

Generalized Continuous Maxwell Demons

We introduce a family of Generalized Continuous Maxwell Demons (GCMDs) operating on idealized single-bit equilibrium devices that combine the single-measurement Szilard and the repeated measurements of the Continuous Maxwell Demon protocols. We derive the cycle distributions for extracted work, information-content, and time and compute the power and information-to-work efficiency fluctuations for the different models. We show that the efficiency at maximum power is maximal for an opportunistic protocol of continuous type in the dynamical regime dominated by rare events. We also extend the analysis to finite-time work extracting protocols by mapping them to a three-state GCMD. We show that dynamical finite-time correlations in this model increase the information-to-work conversion efficiency, underlining the role of temporal correlations in optimizing information-to-energy conversion. The effect of finite-time work extraction and demon memory resetting is also analyzed. We conclude that GCMD models are thermodynamically more efficient than the single-measurement Szilard and preferred for describing biological processes in an information-redundant world.

cond-mat.stat-mech↗