Fluorescence to measure light intensity.

Despite the need for quantitative measurements of light intensity across many scientific disciplines, existing technologies for measuring light dose at the sample of a fluorescence microscope cannot simultaneously retrieve light intensity along with spatial distribution over a wide range of wavelengths and intensities. To address this limitation, we developed two rapid and straightforward protocols that use organic dyes and fluorescent proteins as actinometers. The first protocol relies on molecular systems whose fluorescence intensity decays and/or rises in a monoexponential fashion when constant light is applied. The second protocol relies on a broad-absorbing photochemically inert fluorophore to back-calculate the light intensity from one wavelength to another. As a demonstration of their use, the protocols are applied to quantitatively characterize the spatial distribution of light of various fluorescence imaging systems, and to calibrate illumination of commercially available instruments and light sources.

Confinement-induced accumulation and de-mixing of microscopic active-passive mixtures.

Understanding the out-of-equilibrium properties of noisy microscale systems and the extent to which they can be modulated externally, is a crucial scientific and technological challenge. It holds the promise to unlock disruptive new technologies ranging from targeted delivery of chemicals within the body to directed assembly of new materials. Here we focus on how active matter can be harnessed to transport passive microscopic systems in a statistically predictable way. Using a minimal active-passive system of weakly Brownian particles and swimming microalgae, we show that spatial confinement leads to a complex non-monotonic steady-state distribution of colloids, with a pronounced peak at the boundary. The particles' emergent active dynamics is well captured by a space-dependent Poisson process resulting from the space-dependent motion of the algae. Based on our findings, we then realise experimentally the de-mixing of the active-passive suspension, opening the way for manipulating colloidal objects via controlled activity fields.

Parallel, linear, and subnanometric 3D tracking of microparticles with Stereo Darkfield Interferometry.

While crucial for force spectroscopists and microbiologists, three-dimensional (3D) particle tracking suffers from either poor precision, complex calibration, or the need of expensive hardware, preventing its massive adoption. We introduce a new technique, based on a simple piece of cardboard inserted in the objective focal plane, that enables simple 3D tracking of dilute microparticles while offering subnanometer frame-to-frame precision in all directions. Its linearity alleviates calibration procedures, while the interferometric pattern enhances precision. We illustrate its utility in single-molecule force spectroscopy and single-algae motility analysis. As with any technique based on back focal plane engineering, it may be directly embedded in a commercial objective, providing a means to convert any preexisting optical setup in a 3D tracking system. Thanks to its precision, its simplicity, and its versatility, we envision that the technique has the potential to enhance the spreading of high-precision and high-throughput 3D tracking.

Method for the determination of preferential orientation of marine particles from laser diffraction measurements.

In situ laser diffractometry (LD) is increasingly used in oceanographic studies to estimate sediment transport, particle fluxes and to assess the concentration of marine phytoplankton. It enables an accurate characterization of the size distribution of suspended particles from the scattering signal produced by their interaction with a collimated laser beam. LD reliably reflects the sizes of suspensions dominated by nearly spherical particles; however, when complex particle morphologies dominate the suspension (e.g. phytoplankton) the resulting particle size distribution (PSD) may present significant variations attributed to different factors. In particular, the orientation of non-spherical particles - which abound in the sea - modifies LD measurements of PSDs. While this may be interpreted as a drawback for some studies (i.e. when precise measurement of the volume concentration is required), we propose that detailed analysis of this signal provides information on particle orientation. We use PDMS micropillars with prescribed elliptical cross-sections to experimentally determine the dependence between the spatial orientation of elongated particles and changes in the PSD measured with a LISST laser diffractometer. We show that LD can be used to adequately characterize the different dimensions of the non-spherical particles at specific orientations. Using this property, we describe and validate a method to infer the preferential orientation of particles in the sea. Our study opens new perspectives in the use of in-situ LD in ocean research.

Collective sinking promotes selective cell pairing in planktonic pennate diatoms.

Finding a partner in an inherently unsteady 3-dimensional system, such as the planktonic marine environment, is a difficult task for nonswimming organisms with poor control over their orientation. We experimentally investigate the process of cell pairing in pennate marine diatoms and present field evidence of its occurrence in the ocean. We describe the mechanism as a 3-step process in which pennate diatoms (i) vertically reorient while sinking from surface turbulent waters to a more stable environment (i.e., under the seasonal pycnocline), (ii) segregate from incompatible partners (e.g., dead or different sized cells), and (iii) pair with other partners as a result of the hydrodynamic instabilities generated by collective cell sinking. This is, eminently, a cell abundance-dependent process, therefore being more effective when population sinking is synchronized. We suggest that this selective process, enabling matching of size-compatible healthy partners, could be fundamental in understanding sexual reproduction in pennate diatoms.

Confinement Enhances the Diversity of Microbial Flow Fields.

Despite their importance in many biological, ecological, and physical processes, microorganismal fluid flows under tight confinement have not been investigated experimentally. Strong screening of Stokelets in this geometry suggests that the flow fields of different microorganisms should be universally dominated by the 2D source dipole from the swimmer's finite-size body. Confinement therefore is poised to collapse differences across microorganisms, which are instead well established in bulk. We combine experiments and theoretical modeling to show that, in general, this is not correct. Our results demonstrate that potentially minute details like microswimmer spinning and the physical arrangement of the propulsion appendages have in fact a leading role in setting qualitative topological properties of the hydrodynamic flow fields of microswimmers under confinement. This is well captured by an effective 2D model, even under relatively weak confinement. These results imply that active confined hydrodynamics is much richer than in bulk and depends in a subtle manner on the size, shape, and propulsion mechanisms of the active components.

Entrainment dominates the interaction of microalgae with micron-sized objects.

The incessant activity of swimming microorganisms has a direct physical effect on surrounding microscopic objects, leading to enhanced diffusion far beyond the level of Brownian motion with possible influences on the spatial distribution of non-motile planktonic species and particulate drifters. Here we study in detail the effect of eukaryotic flagellates, represented by the green microalga Chlamydomonas reinhardtii, on microparticles. Macro- and microscopic experiments reveal that microorganism-colloid interactions are dominated by rare close encounters leading to large displacements through direct entrainment. Simulations and theoretical modelling show that the ensuing particle dynamics can be understood in terms of a simple jump-diffusion process, combining standard diffusion with Poisson-distributed jumps. This heterogeneous dynamics is likely to depend on generic features of the near-field of swimming microorganisms with front-mounted flagella.

Emergent Hyperuniformity in Periodically Driven Emulsions.

We report the self-organization of microfluidic emulsions into anomalously homogeneous structures. Upon periodic driving confined emulsions undergo a first-order transition from a reversible to an irreversible dynamics. We evidence that this dynamical transition is accompanied by structural changes at all scales yielding macroscopic yet finite hyperuniform structures. Numerical simulations are performed to single out the very ingredients responsible for the suppression of density fluctuations. We show that, as opposed to equilibrium systems, the long-range nature of the hydrodynamic interactions are not required for the formation of hyperuniform patterns, thereby suggesting a robust relation between reversibility and hyperuniformity which should hold in a broad class of periodically driven materials.

Geometrically protected reversibility in hydrodynamic Loschmidt-echo experiments.

When periodically driven, a number of markedly different systems (colloids, droplets, grains, flux lines) have revealed a transition from a reversible to an irreversible dynamics that hardly depends on the very nature of the interacting objects. Yet, no clear structural signature has been found for this collective self-organization. Here, we demonstrate an archetypal Loschmidt-echo experiment involving thousands of droplets that interact in a reversible fashion via a viscous fluid. First, we show that periodically driven microfluidic emulsions self-organize and geometrically protect their macroscopic reversibility. Self-organization is not merely dynamical: it has a clear structural signature. Second, we show that, above a maximal shaking amplitude, structural order and reversibility are lost simultaneously through a first-order non-equilibrium phase transition. We account for this discontinuous transition in terms of a memory-loss process. Finally, we suggest potential applications of microfluidic echo as a robust tool to tailor colloidal self-assembly at large scales.

Hydrodynamic fluctuations in confined particle-laden fluids.

We address the collective dynamics of non-Brownian particles cruising in a confined microfluidic geometry and provide a comprehensive characterization of their spatiotemporal density fluctuations. We show that density excitations freely propagate at all scales, and in all directions even though the particles are neither affected by potential forces nor by inertia. We introduce a kinetic theory which quantitatively accounts for our experimental findings, demonstrating that the fluctuation spectrum of this nonequilibrium system is shaped by the combination of truly long-range hydrodynamic interactions and local collisions. We also demonstrate that the free propagation of density waves is a generic phenomenon which should be observed in a much broader range of hydrodynamic systems.

Hamiltonian traffic dynamics in microfluidic-loop networks.

Recent microfluidic experiments revealed that large particles advected in a fluidic loop display long-range hydrodynamic interactions. However, the consequences of such couplings on the traffic dynamics in more complex networks remain poorly understood. In this Letter, we focus on the transport of a finite number of particles in one-dimensional loop networks. By combining numerical, theoretical, and experimental efforts, we evidence that this collective process offers a unique example of Hamiltonian dynamics for hydrodynamically interacting particles. In addition, we show that the asymptotic trajectories are necessarily reciprocal despite the microscopic traffic rules explicitly break the time-reversal symmetry. We exploit these two remarkable properties to account for the salient features of the effective three-particle interaction induced by the exploration of fluidic loops.