Marco Cosentino Lagomarsino

How Should Microrobots Swim

Microrobots have the potential to dramatically change many aspects of medicine by navigating through bodily fluids to perform targeted diagnosis and therapy. Researchers have proposed numerous micro-robotic swimming methods, with the vast majority utilizing magnetic fields to wirelessly power and control the microrobot. In this paper, we compare three promising methods of microrobot swimming (using magnetic fields to rotate helical propellers that mimic bacterial flagella, using magnetic fields to oscillate a magnetic head with a rigidly attached elastic tail, and pulling directly with magnetic field gradients) considering practical hardware limitations in the generation of magnetic fields. We find that helical propellers and elastic tails have very comparable performance, and they generally become more desirable than gradient pulling as size decreases and as distance from the magnetic-field-generation source increases. We provide a discussion of why helical propellers are likely the best overall choice for in vivo applications.

Microfluidic chemostat for measuring single cell dynamics in bacteria

We designed a microfluidic chemostat consisting of 600 sub-micron trapping/growth channels connected to two feeding channels. The microchemostat traps E. coli cells and forces them to grow in lines for over 50 generations. Excess cells, including the mother cells captured at the start of the process, are removed from both ends of the growth channels by the media flow. With the aid of time-lapse microscopy, we have monitored dynamic properties such as growth rate and GFP expression at the single-cell level for many generations while maintaining a population of bacteria of similar age. We also use the microchemostat to show how the population responds to dynamic changes in the environment. Since more than 100 individual bacterial cells are aligned and immobilized in a single field of view, the microchemostat is an ideal platform for high-throughput intracellular measurements. We demonstrate this capability by tracking with sub-diffraction resolution the movements of fluorescently tagged loci in more than one thousand cells on a single device. The device yields results comparable to conventional agar microscopy experiments with substantial increases in throughput and ease of analysis.

Concerted control of Escherichia coli cell division

Significance The decisional process controlling cell division is a long-standing question in biology, but the answers were traditionally hindered by limited statistics on single cells. Contemporary experimental tools overcome this problem, but this progress must be combined with new theoretical tools to approach the data. This work introduces a quantitative method for estimating the variables controlling division rate and uses it to construct a minimal model from large-scale dynamic data on the size of dividing Escherichia coli cells. Cell size is found to be an important control variable of cell division, but not the sole one. Conversely, a description where division rate is determined jointly by cell size and time into the cell cycle reproduces well the available measurements. The coordination of cell growth and division is a long-standing problem in biology. Focusing on Escherichia coli in steady growth, we quantify cell division control using a stochastic model, by inferring the division rate as a function of the observable parameters from large empirical datasets of dividing cells. We find that (i) cells have mechanisms to control their size, (ii) size control is effected by changes in the doubling time, rather than in the single-cell elongation rate, (iii) the division rate increases steeply with cell size for small cells, and saturates for larger cells. Importantly, (iv) the current size is not the only variable controlling cell division, but the time spent in the cell cycle appears to play a role, and (v) common tests of cell size control may fail when such concerted control is in place. Our analysis illustrates the mechanisms of cell division control in E. coli. The phenomenological framework presented is sufficiently general to be widely applicable and opens the way for rigorous tests of molecular cell-cycle models.

Hydrodynamic synchronization of colloidal oscillators

Two colloidal spheres are maintained in oscillation by switching the position of an optical trap when a sphere reaches a limit position, leading to oscillations that are bounded in amplitude but free in phase and period. The interaction between the oscillators is only through the hydrodynamic flow induced by their motion. We prove that in the absence of stochastic noise the antiphase dynamical state is stable, and we show how the period depends on coupling strength. Both features are observed experimentally. As the natural frequencies of the oscillators are made progressively different, the coordination is quickly lost. These results help one to understand the origin of hydrodynamic synchronization and how the dynamics can be tuned. Cilia and flagella are biological systems coupled hydrodynamically, exhibiting dramatic collective motions. We propose that weakly correlated phase fluctuations, with one of the oscillators typically precessing the other, are characteristic of hydrodynamically coupled systems in the presence of thermal noise.

Short-time movement of E. coli chromosomal loci depends on coordinate and subcellular localization

In bacteria, chromosomal architecture shows strong spatial and temporal organization, and regulates key cellular functions, such as transcription. Tracking the motion of chromosomal loci at short timescales provides information related to both the physical state of the nucleo-protein complex and its local environment, independent of large-scale motions related to genome segregation. Here we investigate the short-time (0.1-10 s) dynamics of fluorescently labelled chromosomal loci in Escherichia coli at different growth rates. At these timescales, we observe for the first time a dependence of the locis apparent diffusion on both their subcellular localization and chromosomal coordinate, and we provide evidence that the properties of the chromosome are similar in the tested growth conditions. Our results indicate that either non-equilibrium fluctuations due to enzyme activity or the organization of the genome as a polymer-protein complex vary as a function of the distance from the origin of replication.

Isotropic-nematic transition of long, thin, hard spherocylinders confined in a quasi-two-dimensional planar geometry

We present computer simulations of long, thin, hard spherocylinders in a narrow planar slit. We observe a transition from the isotropic to a nematic phase with quasi-long-range orientational order upon increasing the density. This phase transition is intrinsically two-dimensional and of the Kosterlitz–Thouless type. The effective two-dimensional density at which this transition occurs increases with plate separation. We qualitatively compare some of our results with experiments where microtubules are confined in a thin slit, which gave the original inspiration for this work.

Physical descriptions of the bacterial nucleoid at large scales, and their biological implications

Recent experimental and theoretical approaches have attempted to quantify the physical organization (compaction and geometry) of the bacterial chromosome with its complement of proteins (the nucleoid). The genomic DNA exists in a complex and dynamic protein-rich state, which is highly organized at various length scales. This has implications for modulating (when not directly enabling) the core biological processes of replication, transcription and segregation. We overview the progress in this area, driven in the last few years by new scientific ideas and new interdisciplinary experimental techniques, ranging from high space- and time-resolution microscopy to high-throughput genomics employing sequencing to map different aspects of the nucleoid-related interactome. The aim of this review is to present the wide spectrum of experimental and theoretical findings coherently, from a physics viewpoint. In particular, we highlight the role that statistical and soft condensed matter physics play in describing this system of fundamental biological importance, specifically reviewing classic and more modern tools from the theory of polymers. We also discuss some attempts toward unifying interpretations of the current results, pointing to possible directions for future investigation.

Continuum description of the cytoskeleton: ring formation in the cell cortex.

Motivated by the formation of ringlike filament structures in the cortex of plant and animal cells, we study the dynamics of a two-dimensional layer of cytoskeletal filaments and motor proteins near a surface by a general continuum theory. As a result of active processes, dynamic patterns of filament orientation and density emerge via instabilities. We show that self-organization phenomena can lead to the formation of stationary and oscillating rings. We present state diagrams that reveal a rich scenario of asymptotic behaviors and discuss the role of boundary conditions.

Contrasting evolutionary genome dynamics between domesticated and wild yeasts

Structural rearrangements have long been recognized as an important source of genetic variation, with implications in phenotypic diversity and disease, yet their detailed evolutionary dynamics remain elusive. Here we use long-read sequencing to generate end-to-end genome assemblies for 12 strains representing major subpopulations of the partially domesticated yeast Saccharomyces cerevisiae and its wild relative Saccharomyces paradoxus. These population-level high-quality genomes with comprehensive annotation enable precise definition of chromosomal boundaries between cores and subtelomeres and a high-resolution view of evolutionary genome dynamics. In chromosomal cores, S. paradoxus shows faster accumulation of balanced rearrangements (inversions, reciprocal translocations and transpositions), whereas S. cerevisiae accumulates unbalanced rearrangements (novel insertions, deletions and duplications) more rapidly. In subtelomeres, both species show extensive interchromosomal reshuffling, with a higher tempo in S. cerevisiae. Such striking contrasts between wild and domesticated yeasts are likely to reflect the influence of human activities on structural genome evolution.

Stochasticity and homeostasis in the E. coli replication and division cycle

How cells correct for stochasticity to coordinate the chromosome replication and cellular division cycle is poorly understood. We used time-lapse microscopy and fluorescently labelled SeqA to determine the timing of birth, initiation, termination, and division, as well as cell size throughout the cell cycle. We found that the time between birth and initiation (B-period) compensates for stochastic variability in birth size and growth rate. The time between termination and division (D-period) also compensates for size and growth variability, invalidating the notion that replication initiation is the principal trigger for cell division. In contrast, the time between initiation and termination (C-period) did not display such compensations. Interestingly, the C-period did show small but systematic decreases for cells that spontaneously grew faster, which suggests a coupling between metabolic fluctuations and replication. An auto-regressive theoretical framework was employed to compare different possible models of sub-period control.